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Jbull
r t
~
~~ tmiddot l ~
J
l I I
J I ~
~ ~
i
[ Imiddot
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7COHGREso fNTERNAOONAl DECORROSlOtfMARWAEINCRUSTACIONES
11~ iNT_~IQAtCm~JiE~ bull ~ 0 M~~~ cQ~a~pK6NDJ9U~G
CONGRESiNTERNAnONAL DE LA CORROSIONMARfNE ETOES-SALISSURES
UNIVERSioAD POUTEcNICA Valencia 7 11NoViembre19S8
~~
ESPANA
SlllON 1
CorrOsiOll marino Cortosion mQ~ Corrosionl1l(lr1ne
The unpublished manuscripts have been reproduced as receivedfrom the
author~ TheS~ientiiicCQmmlttee takeeMt ~esp~~~abllity for any shy
erroj or olllillsion
Las ponencias no han sido publicadas y eatan reproducidas tal y como
se han recibido de sus autores El Com~te CientUco no se tesponsashy
biliza de cualquier error u omidon
---- bullbull IICHnD~ ~I11)AY 11th NOVEMBER VENDREDI NOVEMBRE --~
SESSION I Corrosion Marine
Friday 11th November
SURFACE MODIFICATION OF CuZn20A12 ALLOY WITH PIRIMOINE DERIVATIVES
Krzysztof Debrowiecki and Kazimierz Boron
MICROENVIRONMENTAL CHANGES AROUND FOULING ORGANISMS AND THEIR CORROSION IMPLICATIONS
E G Bellinger and P Wall
THE EFFECTS OF CATHODIC PROTECTION ON THE MICROFLORA AND PERFORMANCE OF ANTIFOULING PAINTS
E G Bellinger and A J Mitchell
is usually
inhibitors
compounds
an
against
their
of
surtace
to
waters (F)
the IIOst
in
160 -190
001
The specimens Iliad ttle tOnt ot ring~ cut out from recrysshy
tallized coamercial tubes (di~ter90160 am height 200 am)
The grav~metr1c corrosiontest8 were IIIOde using an artificial
see water at room temPerature with contineous aeration
In the preUlafnatY serie8 ~t experiments lIIOClitications were
carried out by p laCirig thespeciiDens into aqueous solutions of the
chemicals specified allltWe and kept at 40middotCtor 2 hrs The
concentration 10-shy -oledJlill 111 the Ibainexperiment the
lIIOClitication conditions were follows
- Inhibitor conce~trat10n 10- to 05middot10-a ole~
-TelllPerature20~060eOmiddotC
- Time 361624 hrs The corrosion resi8tance was measured bY expoeing the samples
to aerated see water for 5 days in the preliminary experiments
or tor 5102040 or 60 daye in the main experiments
Resu1 ts and msc_sion
Only the compounds BTA DAP and BD were found in the
preliminelry experiments to have the ability ot inhibiting brass
corrosion
Tabie 2 The effectiveness of CuZn20Al2alloy surface modification after 5 day exposit 1011 to see )later
COIIIPOIolM
~
bullbullbullbullbullt
bullt
bullbullbullbullbullbullbullbullbullbullbullbullbull
naamptot~rowieCkl nd Kszilllierz BOron
Cul~AL2AiLO ~~bullocJl Inst hit 0 lnorllanic Tchnolotfy Tchntcal UnLHIrSL ty
tiL s-gttucho_JuIIO 25 60-372 IorocCaw PoLand
Introducton
The chemiCal modification ot copper alloy surtaces
carried out with compounds which are good corrosion
The most commonly used inhibitors are benzotriazole and
ot the structure similar to that ot this compound 11 As tollowsfrom a number ot reports 12-4 benzotriazole and
the compounds similar to it are chemisorbed 011 the surface of
alloy to form a stable inert layer protecting 1t
corrosion The surtace modification of copper alloys improves
stability against outdoor corrosion It also reduces the rate
corrosion in see waters In this work SOllMt other chemicals were tested as
modifiers tor CuZn20Al2 alloy (special brass) The aim was
illlProve rsistance ot the brass against corrosion in see The tollowing compounds were selected 110-phenanthroline
13-benzodiozole (BDL 2-aminopirimidine (AP) 2-mercapto-46-dishy
aminopirimidine IDAPI and 2-mercapto-456-triaminopirimidine
(TAP) hnzotriazole HITA) was also used as a reference standard
In the pniliminary experiments DAP +as fOUnd to be
effective corrosion inhibitor This compound was than studied
detail
Ettectiveness ~ Procegtdurs
BTA 293The experimenu were carried out with CuZn20Al2 brass ot the DD 12chemical compoition presented in Table 1 (accordinq to Pol ish
DAP 645Standard PH 67H-870251
1 2
I
I
~ Deteiled experi~hts were iiIodeltwi~h DAP~he most effectiv~ corrosion inhibitor among thecoiaPoUndsstudiEiq The ett~cts ot
Ii its concentration antJ~mpe~ature in modifiction bath on the bull bullbullbullbull T _
Icorrosion rate of brass in an artificial see watet afe shown in Figs bull 1-3 Fig4 pres~ht8 the) effectir~ness of middotQlqMficationvs time of exposure for the saRIPls mOdified Wlth101-M DAP at 20middotC
- bullN tor 24 brs lt
I The gra1Dtetr1~ hur~~t~dtOJCuZlj20AJ2 middotmiddotallQY in the
stationary cQnditfonaevealed that tbecorroliion rate insee water was rdUcabi~bil~ttrtr~a~~ngtl1e brass ~ur1acbullbullmiddottdtil
DAP sQlut1onTlteettbullct otmiddotcconctfrClt-tonofthe DAFIIIOltl1fYing
I solutio~ upon th~ ~~~01l10it~~t~u ~oterY ~l~~At~O)DAP 11 protetedln~iMb~n usedirttbe concentntion 10~ M The
corrosiOnitf~~it~~~a~~~~~middot tji-~iticat10n tibullbee longer aM 10nger1beDAP$oludoMmiddot of concentrations O~5middot10lt
10or~0 tUled at~oejtb~Pdht ~ott~ct ~r increuedthe
J- corrOiti(gt~ ratoflt~OAI~ lllOY~ At t~etu-ra~dtolO~Cr tl14it oclif1cationwlth ui M
DAP swtaot1by i44U~edth~ corrOiJionzte Jbi ntwsmiddotre
lusindelIndent ~t the tioCII041f1ca1iCgtl themocl1iicatlonI ffectivbullnesswasin~bisc~C(~Jtib as ~a ebull Anlncreasein modification tempra~u~ middothllda~sitive effect oli corr~ionate also for D~SolUti~ri otconciitirationoSmiddot10-X bull
For DAP of concentrations 10 and 10 Il1O ledJdI bull nof sufficient iDlProvnt in resistance ~f the brass agAinSt
corroslon tas obee~e4even at elevated temperatures 060bull orr eomiddotc The rate of corrosion 4ecreasedyithincreasing JDOdification
I time and after the 24 hrs treatment it was nearly the slUDe as
that tor uIIIIOC1it iedbi-ass For the DPA S~lutionofconcentr~~ion10-middot the change in
bull modification temperature inthe range 60-80C did not produce
significant differeneilin corr~ionratesThe resultilUJ highly reduced rate of corrosion UCQGlpQred wi thufmodified slllllPles
J remained UnChangedindep~ndentl~ of modification time The lowest rote of corrosion was obtained for the slUDplea
I I
3
modified with DPAaolutionof concentration 0510-amp at 60middotC The protection effectiveness of 97 wae obtained after a 24 hr treatment Further increase of the solution temperature to aomiddotc gaveno ItIOre improveJpent
eonc1QSJons
nllt ~u1t presente~ ~ an be S~~S~l ~n the following conclusions
1 FrOID UIOng 11 compounds stucUed the Hst corros1on
protectioopro~ruH h~ 2-mercapt0-46-timinoPii1D1idiM (DAP) 2 The protection properties of DAF 1rovtd with increasing
rilodi t1cadonterature
3 DAP wu found to tie better corraaion inhibitor tor CuZri20A12 alloy than benzotrlazole
The lIOdit~cat1on effectiveness did not cbengewitllUme of exposition ot 8amP1es to an artificial see water
Ret-nc_
1 SpinellLP~middotMaj~hM BrunonoG Trabanel1LG r~t u
KDrro 31 919-923 (1980)
2CignaRG~oG BrCOZrobullbull J bullbull US 223-225 (1979) 3 WalkerR Corratoft 2Q 290-296 (1973)
4 Trabanell1G Zuchh1F bull BrunonoG Carru1t1V t Korrobullbullbull 24 602-606 (1973)
4
-- -II
JII
if ~-it CKpirons 101- dltf1lresect
it
it ~
coyio~rOIl tit Ctn20A i ii I uy if bH 5 dys ot
exposure to ltir tltl Cl d 1 see water ~ff~CdtioH condit 10IlsI)II1
it 10-middotmoled1W te_ 2o4Q6080middot
fi92 Therote of corrosion of CuZn20Al~ 01 aff~r 5 days ofit ex~EJuHl~ortif ictal 1-eet wateYKocfificiltrorP conditIons DAPit
10-middot ~ole~ temp 20406080middotC it Fi~~TIu(~~t ~f ~orro~ionmiddot~f cuiP20A2~111i) otter 5 days of it ~~~~I~iii~~rtIti~i~l s water Moaiflcafiorl ~ohifitians DAP
~ 4 ( 1- -~~ ) ~~ II10-~ IIioledIII temp 20406080 C I ~
Fi~4 The lIlQCl~t ica~iol) effec~iven~s Yt exposure time torIt CuZn20Al2 alloy in ortiflci~t slaquolelfaterbull KoclificaqQnc~pditions
bullI it
~ Dllt~i~lQmiddot~Jel~ telllP 2PmiddotC~l hr~
ibull -
bullbull bullt
bullJ
bull bullt
bullbull 5
bullt
~ i ~
12bull 5 p
3 6
r1g 1
t bull 2OC iIrC to-c tIOC
-24
16 iMtiMtillle Ihl
I I
~ N
t ~ 5bull
JOC bull AOc bull IiOC -we I
I I I
3 1amp l I modifiCation tiMe Ihl
I I
t
f ~
_--
yshy-~~
-1shy~~(~
Jl -
- _JIJ 11 _ bull ~ bullbull L IIIJIIM
I bullbullbull shy
-
if Ilt
JII l3$i]J ~e-l~ ~ 1
J ill KItt
Ii I 1 8 Ij j iJ)Sj
emiddot Sllll0 r B 8Je Do Ji
t ~l u Ji ]
1 0
j
j
r 1l3iiiJail1h
bull III
~ i J ii i )1 t~I ~
~
~ )38 ttl i UN ~I3 11 I ~iii bullbull I fi~fi IelI M 6 6 bull s plj i I Jjhi
~l -u II11 i~ Ji ~aJ j~ ~ tll
- bull
-------------- -- - --- -- - -- shy
2
This paper desr1blS the use of ti1c~lctrooes to measure pHbull bull _ bullbull ~- l bull bull
iJQ2 chmgtS ~~ 1II1C~olones ~middotilIlg~ ~f~~s i~rsed ll ~ter ~ wlJlch t1)U11ncjotgaI11sllls are grolog SOllie p~tnt1al 11P1~~tions C1f~se cIIatlges to corrosloO at the
lt I )
surlP alllO br1lJfl)dl~ - 1 - ( _ ~ -
ItIiterWs 8nd ~ ~ ~
() poundI~~lt - -
Ttl lec~_ 1n theu bpelsnts hIM bnndeScdbed In
~~~I )iel~~bull~~i~) but n 11 ~ haJe
- Aj ~-
(1) pHel~
ThapHalec~ lII8rII basadQIJ tne-iWof It1tQ (1__)
wre the pHlt1V8recenton of~~l~ 1a fiI)OS8d bull 1Ihart po1nt extlndlngfro- tt lnsulatlnQ (Flljj1) The
pH andmiddot alUlllno~nlcate glau 1cropipettesued lnthe - - ~
electrode constnction went ~ using bull wrt1cal alectrode
puller assntielly bull vart~c81 version of the equiplllltnt
orlQlnally developed by Al8Xllllder and NasWlaquo1953) The ~ ~ -
electrolyte u-d1llllS 1N tel and filllnQ IIIIIS 8aoIpllshad using bull ~ - or
aydnge 8nd a vary fine draIIIn out Qlass capillary
Once f1lled the elfCttocJes were soaked for severalday~ in 1111
tel to allow coqlletehYdroi~s ofthe pH glass lin lnternd refelflf1(e of sllver lrecoated wi til sliver chlorld~~~s
ccinmctedmiddot to a gold plated pin iealed into the imd of the
electrode lII1th~ill~oo rubber COMPleted ei~trodei er~ stated Up downIIIards lna dilute solution of ~r~iIcrcj In allmiddot uper~ middot-temal reference Qf bull~~l
allCtradilllllS UIIIId DurlnQ -~ts -n bulllact~damiddot was c~cted to ind~ ~~F~Y(_ itrgtbull The cStcu1t layautforbotitthiiptl lIlIdpDz electtaifes
~
1a IIhIWi Sn Figure 2 bull
~ -~ - ~ -- - - - I -
CaltbnUariof all alacbodn id Clirr1ed middotOutmiddotbllfGn ftc AIIICPUIIIri inmiddot c- elctmdeiimiddotmiddotmiddot_middot~
ckalng en apars-nt by in nJIalt~ orthey dldriat
pEOCia ~lbblIt ~lntal callbraUon In ~
CIIIIi8S all ~ datafl(WtMI middotmiddotdl
(11) 0ItygIn a1acUocfes bull
TIUI catheter typa elachode use a fiat ended and of duw 1 to 15 _ It conslatad ofa ~ uia P~tJft- wlra
aeelad 1n 9l~ and IIIDadded in 1IPaKY~1o 1I1th a aUler wue KUng a raferance (see FiQ3) Ihen the epOxy hId~
h I of h l~middot careMJy ~ flat to UpQM
bath the plau _the ~lwr 1IIIIIbnre WIis ttn8llPlled
ecrosa ~ flat lind by rep8atedly d1fIPlng Jt into a aQ1utJDn Of
collodion 10 atheta1cohol A Pola1-1slng voltage of m all __
applied bit eachelectrodl and these re not thariuSed for
axper1lllental purposes until middotthe response I118sstllblallihen
bullp
4
bullit tuffer elut1on a~ a eorustllllt
it i)
~~ ~ ~eted IItld accepted for uSe elliCtlodes
it lt gt ~ ~~J ~J _ bull~ - - -gt bull
IIIIiIr~~Jn~IIOWt1on of 02 KCl D~ygen Ilectrodeser _iliry~ ~~)~ 5middot -1 ~ J~- - f -
it cli~rated aga1pst solution I of known oxygen content (as 1 w f - w ~-l _ bull bull
i IllllllaUred bY n of the l111nkler titration thad APHA 1915) ~Lmiddotmiddotlmiddot ~ ~
it Any el-=trodI not lIlCtIib1ting a ilnaar response rejected ~ ~ ~ ( _ - ~
zelia axyg8n lution obteined by the usa of a buffer
i aturated wUhpyrogallol crubbed nitrogen The 100$
~~~ reed1ng obtained by bubbling a1r thlOUltl the ~ ~~bull ~ ~ bullIII burfal far 12 hours at ~~ture 8M pressure- ~t bull
(b) [)cpars-ttal~
All -rs-tts IIIIIl1I Wried aut in atellperature controlled
bulli it
tank pliad an a teflain NYler ted l~ steel patfon ill insidil a ane _tre CIbI Faraday CIlIIIi(fig 4) Tbe~
prav1decI IIf1th a lJrfQII hinged door at the fiant for eccns and[f as gro~nded to earth to scrbullbullnout radio frequencyi intelfuwce The _tar flowing ttmluitI the experluntali - tri pullPed at a controlled rate fro a temperature
~ controlled reservoir For all experillanta involving algae
i llllA1nat1on provided using daylight fluorescent twes if l1d teel and 1Ial1ns plywood plate approxilaately 20 bull x 15
wire Ixposed in the -rine environmantfar several IIIQIIths to bull
allow a heal) growth of IIIICrofoul1ng organi_ to deVelop The
bullI steel plates had been placed utldally and IIIItre exclu8ively
bullt
~~==1$ and ~klOOrganl_ ~ldirig Itltena ~srar-nt of their
OWl The wooden plates 1IIIIIl8~sn the Jntett1ifalteclJQii
-f llllllre COIIered lIIIith ta 01 tt- alp InWfCld1IIPtth
occa1a1onal bryozaens There IIIIIIl8 IWIo lagII quantitibullbull of
~ retained -gat the algal fu-nt nPiate lIIIIIle alsO cleansed and stalJil1aad_ bllton ~
provided On 12haUrly~ ~dllrUght luar~w1 RddtUOlllll steel plat8s ~~In Alii ~dIxk~ lit
Uwrpaol (lie) IIIhare they IIIIIIl8 co~ by middot~Of~
(xtllus edulis) and the lISCld1M Clona intUn1l us
~1IIIIIl8 roved f the pJaqa NP~1id In cl_
ta tor 5 days at cansqnt ~ toIitDUampe
pH WId pOz ~1ane frc the extuMl t~~
the fouling to the Milstrata llllllre ncarded tt elecUadM
middotheld in a rigid pttllpex ~rr1et attechld to II ~ly
Jointed GIl on the lI1crcIIanipulAtor To nciad the muM the
tnraugh the foulJng tI1IIIIerds the ~ta JI_ta 0( 1 _
could be acCUllitaly controlled by talng the tIM clrlw of
the n1pJlator Iftn each lllc~ the DVIII ~oIdlft9 shy
allowed to _tabIU for ane wte prior to ncordJng the
VGltge~
~
bull a
-lt 1
l~to the_ batnacl~ 1ls~ COC)trol - fmiddotmiddotmiddot
recod4l9sof W pH 8I)d pOi at the SIJIIEdepth outside tOe 5hell middot-middotvmiddotmiddotmiddot_I~ lgt~middot middotmiddot~_middotmiddoti 1gt )
wete made
-flesolts ~
(a) pH IIIldIDz p~lllllS
The _re~ts for Hancl p02proflle tluough both algal -d - ~ gt Comiddotmiddot i -
eniIIIalfou11ng are gi~ in Figs Sto8bull --
TjpicalProtUeis daIiilttuougha t of Entronlorpha are ~1n -bull l J ltl ~~ -
Fig 5 - lIhllsttyp1cal d1innal fluctuations CICCUIJ8d during bull ~i bull bull e bull bull
photosynthetiC- II1d non-phOtinwnthet1c lods (Fig Sa) mad _~ d q _ _ __ _ i bull
decreases inpHlIIIIl1I~ IIIIa) fro-theaul-f1lCe of ttWt bullbull bull ~ bull
dcyenI tqwards the abstratacn tdch it enclOSlild (fig 511)
~fal1s1n pH--nted to as-ach as 2 units Over 9 IIIj
dIpth
figure Sa ahoid the thangea in pH -d Pl2 between 11ve barnacles
with depthln a stlat1c envirornent A steady decrease in ~
occurred as the plata IlUrfe~es toiereappro8ched pHWtially
fell on one profile before rislngbut lOSe steadily with daPth
on the second praflle SlIdlar changes are IIt10wI in fig 6b
with depth into the shells of dead barnacles 80th oxygen -d
pH levels were re~at1l1E1ly steadY ~ similar to the 8IIIblent - ~- lt
envlrOlWOt Until the -actual dead shell IIias entered When a
IIIUked drop in both fJ02 and pH CICQ11Tedbull
~llell in rigs rit JllellllUrellt$IIere maltle ~
iOlfillidual IlUs$els withtiRle (Fig 7dlb) In thll first
experisent elactrodes were placed indicated (fig)
arCUld indivicUllalsln genU flawJog ~ter The
resUlts show bull ~tle fIwrease ~ IID2 eMIS bull aiJc hour 1Ucj
The pH~t of the~ II4ttIn ~~atmiddot~
81 (te UlbientpH being 80) but 1GII8 ateIIfIU ~ the
1rtIa1ent aJphon to a6bull 111 iIadIIr ~~thI aff~ of
flOiW 00 local ~ciIndltJans~ the ~
~tadbut tile flow at~ turned Off tar 25i1aur
Jadin thelllddleofthlt ~The nailts g1R
in FlQIb -PD2 IIMtls mae ~y inflQldng CCnd1tJanamiddotbut
fell -zkadly fter alhoit lao w flaws caadawJnit a
~ fall lit 90 alii Levels rapidly ratond onca t IIIIUtchadDli agaJil No ILICh teed varlauon ~
in pH around the eldlalent 1pbn 1iIIflImI a taIdyJncn4IM -
obsmt8d aspec1elly cleM to thIIen1Ml CondltLni around
indivOJa1 ~ lIIhllst of interest are lMa repl_tative
of field conditions fro around groups or clIIIPS The
IlICpIIr1ants were ~ ~ted Wittdn denSe clUIIPs of ala
fouling a plate These r8SIJlts are given 10 figs tie (1Il1Ud
IlUlIS8ls) -d 8b (within byssal thrtiIds) In tatic ccinditJans
amprCUld the _15 u-a1(8$ ~ fellrapldly (45l) reelJ pft
rose sl1rjltly and then feU aftar 2 h llhen flowing condltiors
were resuraed after J hthe ~ rose to the original middotlevels shy
The pH rose lArlcedly at the eltictrodenear to the xhalent
bull bull bull bull bull
bull11
bullbullbullbull
it
I
8
s~ (3) but reriialned stlaadyahd ultilllateiy fell silghtiy (2)
inttie genetal fooling inassbull
~
III1thihthe1JyMli~~ Or~selci~5 IiIfieretilltti($trltus teniis ltO -=IiUlteliter~ ~ ~(Flg 8b) ~ lIIithrrolaw~ cCincentreuoos diCIppea framp1~D lBigtif3S
~bUt toaitnpl~lylt4i 150 IIifIo lIIhfjnficw t8st0ted~middot pH
t wi~1rithi thread ClUIps~lMl1BtO 74 (3) IinIt B~ito
785(2) lri-staUc cinfit1at IoihUst ettn88dgtiofthlt ~
it -flnedf81r1y~at aroOOd17S~ lIIlanf1Dlilwu
rellitoteilpft inltleUyJose (by 04 UI1ts at Ii1ta3) but it theo
bull fall only to startrfs1riq tgi1nattM not theIlXper1ilent~middot
bull Dlseusslont f tMresulblmiddot obta1nIid it ~be cl8lirly sean that f(IIJUngI
bull species vethe abUlty to Gtly IIOdlfy thdr loclmiddotmiddot
bull InIl~ts
bullbull L1t1t end dark period tluctuat1ona in algalts IBre obsarved
end ware lri close wpeeIIIen~ with those reported by Terry end
bull ~
Edyv- (1981) with 1ncnasaln both pH end IID2 llIrJngllttt
traquor1ods lIhilst thlshappened ttM surface of the t pre
bull fU_ts ware frelyprojecting into UJe water -deep intottie
mat nearer to the solid _stratlli whl1lre shading fll1lli light andbull accuIIJlatlonmiddot of deconIposJng ~gallIIBtllrial DCCIINed pH valuesbull fell IIIBrkedly Concurrent wlth this dlQp in pH ilaS II fell inbull
bullbull
9
oxygen Such t~ral and $PiItial va11ations in ~andpft
would certainly qive 11se to corrosion cells middotatmiddot the sUbsttatie
surface ~ly resulting in iorroaien pitting
The r] _tabellc ctivlUee or fau)Jng -=raJrwerttmlbullbull
together with their prapeN1ty to-=ci-1tritla~
tlwlsalves wIll also lIied to 1dcIoenv~ lIOdifktIona
Resplretory activities wU1 tend to ~~ tn-the tft and Input caapanants auch a cinia~nY qatk
1mIertabJatesmiddot ara mlk 1bullbullbull middot the bulk of their nlUagllnoua
wastes ~ excrated as ~ 1heexpftunt ncord~ pDz
end pH cningS amN 1nd1v1Q8l~ ~ ~ri
claady 1nd1cat41 the ~ of _tft~t GIl tt ~ of lnIirollliental changes Inmiddot expr_~ta lllhere no Wlter
--nts occurradthmi a np1d dacreUfi1ll pDz
COIJIdalo be a ~decreae in pH (no a) v fladrlg
condlt1C1ns are r-ed tIwn Ieymiddotmiddotor iIay nat be a 1bIIeIqI1int PH
1nctease dBpwIding ~ the --- ofmiddot iheltr ~ by the
organl1n the cluIIp 0wIgn wui a influenced by the
proxJa1ty of 1nhalent or ~a~ the dlnsity IIId the
tIlJcknes$ of the clullp w111alsa t)IMt wiuence in ~
wetr -N_tbull Hencein thick clampllP aUt and detritus
cCUlllulatlon is are COIIIIiIn ~ theh billa IIIdfbls
restricted Br8IJIIltdow1 gf organtc datrital _ter1als CI3IIIIUIIS
oxygen resulting in recLlcing conii1Uons llleasurlllenta haw
shown that sulphids qulckly build up In such deposits and
sulphate recLlcing bacteria y tie-~ SRBconcentrtlonmiddotin
J -I
II II IJI Ii ill
II
-bull I
11
10
the depOSlts woUld terid to Ile lOW at We start of ~r1ments as
condit1onswere nat favourable (POstgate 1979) The cantlooQUs
prockIctiPn of r1th f~plieUdofeacal_te1alor IlUlpllonill1l
~ts bYpoundilt8~ (~~Iick1ngand 1IkIod1) hOweva ~ t H 1 (
IIIOUld rep1dly altAIr theli11l~t and the rua~ofSRB re
notedtofncreasemiddotttcllll ( 1 SRaIll1taat llaUt 1 x 101 SMIIl-1
aftet bull -th
IUctotJlIIl corrosion canfe=t _ta1sln a1Ullberof Ways
(Iverson1972 fUlllIt end King 1975f 1Uilr 1981) pH can
aff~ the ectivity of the 1IIin organlasinuolved the H 1IIho
hava tin qlu pH range beban 10 12 (Graff 1981)
although tIdS -can be lnf1u8nCed by ather fllltors In atI1tl11n
they lIKIJlre a catbon end nitrogen aurCa 11 various
salts tin alactron accaptor (HedJart ~ idbullbull 1985)bullAil of
U- facton are availabll in the debris -alltting b41tween
ar1na foullng -d the 1lUbstra~ 110 providing bull potantlally
corrosiveanvirorwent
Evan IIIhete such accullIUlatiori does not take place varlable
changas inoxYQ8ll and pH ~trations acrossthe suface of
teel soch as C3I becaustd by foul~ ~ C3I give ti_
to corms1an calla lalildlng to piftlng corrosion
There is an initial tllld8llCY for the pH to rise as a relUlt of
the excretoryactivi1i~ of fouling aniIIal and this 1amp UIIllkely
to give rise to corros1an problems Eventually ~er feacal
h
1
~I
materials and detrltus~ill build up around or~anisms ~re bo~
low pH and p02 devul9ls providing a potentially coJJOampiIu
envinx-nt flaw does dify thh _build UfJ but inmiddotthbull
Ixparlunts carrllldotttid notprlvl(lt it R~~r it
l8Iilated the depth of deposit ~1~~ in t- ~ It ahoutd alao r~~- in f~ population so-1QIJvlWela 111111 be dying or dUd Thlb
~lng~wUl~1Ielacal1Md conoa1ve ~iccandltiorJs-t SR8a 111111 b4I fCllllfC314lng ~y pOIi1desa hlbitt~ HtInd 1tt~ WIi~ul create 1~1Mct Jrcx-ntal dlffarenca ~ ~th l1kel1haOd or pitting
n
References
Albilnder J~T 1 bL~1953~~(9EthII prockIctiPn of IdcroaltJodIIs UIIId in I~ studies Rev Sc1 lnstra ~ 528-531
_r1can Ptbl1c Health laSoc1ation 1975 5taIIdIrd~for the lIIIIIlystS of _tars end taatms 11lth pounddIt1on 1IPHIi-~ 1015 ~taenthSt NW 1IIa-l~~OC 2IJ]J6 USA
Bau-8laquo=k1ng C~IIIG amp 1IIoOd EJF1955 Biologlcal ~ in theestuarlna envi~t 1 EcolClil) of the ~ cycle Kcnlridlkjl Ned Akad -t ~ Ser B~ fi 160-181
cter DB amp SUver IA 1961 In IRafaranctlElectrodl$l EdltolS Iva DlG and l-z Gl lc8dilll1c Pr London Maw Yodlt Chapter 11 ~
Davies PIII I 8r1ric middotF 1942JII1c1oel4iitrodas for-aln local oxygen tension in n11 tissues Rlv Sci InStruII Yo 524-533
EdyileariRJGJ ~ Tetty LA UJ83 The 1nfluenc ~lIlcro algae on corrosion ofstructural steels used 4l~ the 1tIirth
Sea 81odeterioratian sect 336-347
bull bull
~
bull - -- ~It Jf bull
- ~ 13
12
Terry LA I~Ed)VeangtRfiJ 1981 lIi(toal~IOI1CGrmianF~ l)i 1St1oJhemailnEifooiiiigffixed gftshoreIt Bot 1Iar ~ 117-183 lsect~~jlaUoQ1l1 _ ~t Ef)e~gy Offs~r1I Te~lOtjjy Paj)8r
-middotltcntpI)~it$t~l91-16PP~middotmiddotdl ( I
bullbull
iIIalk e1 JL 1971 bull Ion specfie Uquid I~~ e)(charig~ ndcroelactroclels Anal ChIIaI ~8S-Q2 AGraf1~ w~S 9S1~ 1orrosloh InFIntroooctI00 tooffshOle
51~weSoes~~fab~lcat~onIns~allat1on Gulf Wall PI aelUnger EG 1982 The appl1~tlon 9fMl1ikIngmiddotHoust(jfW~Q23g-258middot icroelectrade in the study of pH end PD2 in lIHinli
HerbierC SN fGH~rt~ )RI -Stoctcdalpii Watltif1Sal RS ~1ronIIInb Ub~~ 27l-Z79 1985 Fac~or lontro1l11)9 tha activity ofmiddot sulpMte
bull Wall P 1983 The appllcatlon Md ~ ofiwrJ)e~c~odl shy-taicfii9 tiiCtetili tn~ diiilncj tar infection
techniques for the iiaurnt cif icraeov1ror1liltital5QiCiety ofJebo1- Engi~ SPE 139781-10 _i~~ - ~ ~ i ( _ IKIdif1catlons caued by aGII8 rlnefou~lng -C~ebullbull
PhD lhaaia Ibiverslty or ~tar UKbull bullbull bull -
IIH1nke JA 19fiSa In EXperllRants in Physiology and middotampioClIiIiiilatJift l c E(If(orltGAiKetkutgt ~c-Presa-
Wooltdngton AObullbull Davenport J 1983 pH lind IlIl2 IMla ffl~- j~t~5 -lMIIleath 1IH1na ro-fauUngGqWl J bull Expbull 1
B101 Earlsectsect 113-124 Klnke JAIII 1969b II) Glass IUcroelectJQdes poundx1tors Uwal1~Ui~ OrOllt arid~t Nt JOhn fII11ey YOI1lt thaptar 11
~-f _f~ ~ ~~nt ~ Igt
bull Iverson IIIP 1972 Biolog~ Qrroaian In Advencesin
corrosion scianca and tecIYIology 2EdS FCntaia G~ I bull IIS~et RIIIbull p~ PJaSsLancIoIl 1-42
bull
Jorgensen BB Reusbech NP I Cohen T 19B3 Photosynthesis and strUcture of bIIn~ II1crobWp MC1OIIlectrocki and ~ stud1ea of four cyanabacledalt -~usllWtOl ~bullbull 1~f093
t l~ O~ ~~~ H~ ~9s7~c-gtHatste~iecfn1k van shy pellediftlrtenPt - Stlchelektraden (1-Su
AiJBitiKlificfiliilislflilr)zlIr polrapusdiailJIIissui1g_ liIserstoff(jruQkas fur d1eB lIsUbullbullung der IIliJlttOUf1cillatlon PflftgaHArdI ~fl39
bullbull Mller JOA 1981 tals In EconcinI1c II1croblolooy sect McNbUdbicldatariOtationEd Rosebull A middotAcadelc Praulondcn149-202
t Mller J DAI King RA 1975 8io$~iorat1on of lllltals In lIIi~1al ClflPlCts of the deterloration of tarWbullbull EdsGilbertRJ~ middotloveloCk 0111 bullAcadeic- Presst IILandon 8Jo103
bullt
IPotgate JR 1976 The sulphate-reducingbacteda
Callbridge tIn1versity Press Cliilbrldg8U-K
Reusbecb fUI i Ward DIII 1964 i~~~1Ida5tud1el of
bullt inter$Utilll IIIItez dttm1stry and photosynthetk activity
in ilflaI spring Idcrob1altAppl~EIi~ MerobW tsect4IlU-27lt I
IIbullt bullt
bullbull Ibullt
1
Inmiddot middotmiddot11middotmiddot f~~ a~ -tmiddot bull J~f
I~ W
i5iImiddot J~a t
ilmiddotmiddotmiddot I middotIt Ii
l
~
middotmiddot~~middotlL= g r
Lj Ic
t
i ~
I
gi II cEi ~ I I
1 t rW
D
tR bull
bull t 10
11
r
i
I ~
j
~
I
~bullbullbullbullbullbullbullbullbullbullbull
LV
FigS pH and p02 profiles into macrofoltling eomlllllllitilS dominated by Balm trenatU$ on steel plate 11 two pH profiles Ib) two lIOai profiles leI pH and P02 profiles into dead barllllCle shiella withdetri~Us I I
itIbl r p02 ~ I
nvnHg 11 70
j ifJi eo 1amp0
laquof it 30 s
I 20 N
ca 11 10 a o -- shy i bull
11
80
19
781 o 2 6 8 10 12 0 2 6 amiddot 10 12 dIpdI Qf ~iftto fouling In HighIst numbIII pIatIi
8deg~~
~i c5 iioo5 fiil
pH 8 bull_ ~
~~ ~
_ bullbullbull- pH
-pOz
(cJ
e-Clf ( j~JJlaquo1- --~I ~a ilil - _ I i ]1
~ ri en i ~ oJt ~iJ id tUI
2u1U I shyI t
shy1 2 3 6 6 7 8 9 10 11 12 i shyDepth 9f electrode (mm) into fouling
1 i
1
~----------~-------
~
~-~ lt1~
middotmiddotPmiddot ~
I
V
~
ill
1 Tbellitt~~ of cathodiC protection OO~~lIic~tlO aridrfOflllUlc
ofi AA~Uoullr14l ~1~I
I
ael~lner EG middotIUtcb8l1AJ Dept lt4 lInY1roMe1lal Bl01011
=~~venilt7 IlariCheW 1f139PL $ ~11al1df
r
I
Ablitrict ~~~ e~l billttI ~ted with ~pert1n ant11_1111ampp paint llere itpoii~1n _vatI- In the tl~ld iUIit 1bOioatbrt~ScM Were
1IUbjll~ to qtbocJ~C ~teltt1on byen _ current (~)SJi inci EDAX Iieamp~ta IJdlctecI ~ lachln teoc copper OIl tbePlCtec~ pa1n~ atter14 dQ1a~ protected Pllnt abo ebowed
) reduCed bacteiial activIty butmiddot ~e4 dltoe Iu1d protOzoanmiddot co~~l~tlont~1l IlU l~byenCOCltact not byen dUfwIl00 ~r _baRl lrillolvedln tOXln nil ereio noted
l ~~
1
I
Introduction
Applications of protective coatings and cathodic protect1on(cp~l
are recognised corrosion control methods for IIIBrine steel Itructureso Cathod~c protectlOll preventl conCIsion by Int~lng til8ctrl~l
currentS trem externalllClUfCa to e~~tthlJno~ e~ carrollon feetlans C04U lUChu tUauJ1ng Aelnts a
(
barrIer to ~ the flao of couos1a1 ~aboIHnu~r
Dccurrlng anodes cathodes or wlwit gelvWc CCIIII1 ( 19681
flinger 1980) AnUfauling ~ also JJICOqIOrII~ b1oc~~
Tr~ylt1n nuor~ (~) end o~ Oxide (~) (~ flJ1J
- of u blacld1111~
V _Ut Evans 1981) ~ ~
end growth of bipfoullng 0I1I)IIIII1IIIIe Wdd~ ca ~ of
paint rusendmiddotmanemiddot ~(r~ til )
As corrosian ~t1on ~ bICCIIIII -an ~1tlQd ~ of
antiroullnQs 1 conjunction _ith CP have bllClIOr ~
resultIng in reseudt 1rwestllj111tlng trw COIIIPIitlbUlty or VU~ ~t t _~
rOratlation to dlfrert levI ot CP (~Eelf 18110 ~
~t 1980) RelatIvely 1nIestiljlllUans have ~ urtw ~
or CP an thIt anUraullngpert~ of palnta
In thIt preseot 1nlIestioaUon tV~iln111 ronutlon ud
cOIIIHrclallY incanjlntlon wIth CP was ~t4IC~ Iw I sraed current (-966 l1li) arid exposed at the 1 Strait IInQlbullbullbully
~t1ons weremiddot 00 the IIhcIrt bnifects the PprIUd_
had an a) the various COIIIPQI1eIIts of n pe1ntf1l11 end b) l1oft~
developaentbull
bull bull ~
~ ~ - f
bull ~ _T~~-middot~ ~~- ~r _ -$1
(1) ~rlttionor Painted Steel Billets middot_ ~~tmiddot_tlJ middot~hmiddot~--~fmiddot ~ middot~A~-~~middot - -~ ~
Thirty-al ateel blllets (8-10aD2 end l depth) were cut frollbullt ~middot~middot~middotmiddot~middottrSmiddott ~jS-~ It bull gt~ tfl - middot--imiddotmiddot ~ t1dItrd steal plates The dimenSlonsof -thB butits-lim chosento
bull -~~ middotmiddotgt2h ~ ~-Imiddot ~~rt ~- -middoti~~ ni ~~middot1 l middot-~t~~middot _
(a) fecilltate amiddot Ie detrucUve -thxI of prePraUonpdor to ~~_gt gt~ k7i ~_ t~ 1--1- _ ) - +--_~ -_~bull ming electron ~ 4II1CI (iialloWdirectobs8iVatlCnS of the
- i lt-- ~ gtbull -o1e billet 8Uface
gt r~middotmiddotj ~L(~middot~middot rmiddot tmiddot J ~ - ~ - lt _ bull Single non-standard (1hIula~) nickel lIIireSmiddot Spot Welded e1Cing
bull ~1deJ ~ b~i~ ~j~~ tri afl~1b1ipOi~ f ~1t1 ~4middotmiddot f-gt -~ ~S-- ( -- - - _bull resin EactI billet then Undb~ tI1d prtiladlllltha5O __ eyilr
bull ~~_ ~~imiddot ~1iil h~ intlfcili~ ~eonbt1nlyi Tr1butylt1n F~id(18n)~~~O~dettu20i~blcicld8is 1IIfii bull i - ~ ( then 1IPI)l1ed to glvaadry Ulil tIi1c1ltnOss or 1(1)-150bullbull
tbull (2) IipreSSeCi Current Syat
bull A rigid ~tic due 1I8S drWedllliihwrtv-s41 liiIihOieS IIPProX~
bullbull 05-10 cia episrt ~ Pilnted ampUl8t Wsfbed IntaPQSIUon ampy
threadlng the nickel IIIlr8throu11 the dinto a tarproot junction
box Eliltilen of th8~1l1ii (recelving ~t-d curnritlere IIIlred
Into bull block connectOrrecelvlng dc trOll is potentiOstat The
rainJng iontroi blUets Wet8fbedfl11i1iiy lIIithln thejtlytlonbox
i~ityd~c IkiliPly ~~ ~lat1ruaWiode 8ncIacalOliel
refer~ electtodetoiere fllied ~ the per1aetat of the plut1cdlc IIIId
connected to the pot8ritlostat The desl~ of the potentlosbt ailoiolEld
~PlDteItlO1TClJrnnt to be MIllntained at 8prampset levtl of 966 II1II
c
r~
Grid raf 1J5s-t 114iH 563724 dud 0 Juna
Alolecule andbechr1a in general are ~ to
sutface lI1eroleyer (nlUSton) of _ter (~bullbull 191KJ)~To avold
contaJnatlonof ttw painted surf by U- -vents
blllets Were s-nact in a 1 conte1ner or atc1le (02i ) f~~nd
ter Iih1ch then f9ICI in tv 1IIhere the b~
be r-oved undeNeter El~~ bllleta fa Mcb trtai bullbullel
removed over a 1 dtq (336 hr~ expoeure perioda
each being ~~bn uposurnof 1 5 11 iA 168and DlIIauz1r
Arter field axpauHli the billets wan
fUtered _tar to ~ 1_ debtlflMd far 10 aJnuta ~
wv ~~~endattid sdbUiUd(otO
~ ~l f~ nn1ng el~~ --
lOOted onto 1middotCIi ~bullbullWIsend carbonmiddotcoa~nmiddotmiddot~middot f
each billet Ws~ Iis1ng a ccr1dge ~el~~
(5360) IIIlth anergy dJ8perslve X-ray alcroenelre1a (mx)
_lysis the les Wen reNed end coated III1th vold to al]Ql En
detailed sat of the alaquofa1l
(4) Lfiborattux ~
Painted taa~ bUletalllllre pnparad ~
in a Mrlne reclrculet1ng flOlll _ ~
f1Oll the ~ ~a1t~y Tha~~Un bullybullbullbull
10C M1nta1ned ~~t1as of ~t~la protazOllbull~bull10shy
(Entermap) ~ ~~~middotNlt~Eamp1~ __~and ~ r - ~ -
certaJn ~lc 1IQ1~ 1(1 the
the painted
~
thre repl1catft fn
plKed 1nto(022_)
- thIine1J~~ -lt bull
Atter mAX
(1-2) and i-n-t bn collected
14OC
Pain~
-
12 daY~ exposure and prepared as in d)~ - t
~I ~~~-- i~yen 1~ v
Rflltsi ~ _ ~f-~lI ft j bull _
rt~ll0~~~llY~~~~peln~stetsl~11i~ex~sed for
y~t1~J~ ~~~~1~1~~1UII~ya~kooJ~~~lt ~ ~~_~~I1J~ ~~~tomAX -lysis ~era than gold ~ bull bullbullbull c - _ bull lt
~~fllr ~~~ vllUll ~t1ans of the rfee htures and gt Co
~f~ - ~~~ ~f ~ ~aobta1Mdue IjIll811 iI1Flg 1~2 The IIIIdn ~~ ~ lt)ji9 )tmiddotmiddot - ht
- bull
~I rIIplt~ tIt8 -~ cOlllponanta of the pei1t 1bullbull WelIIlItr1x $ilj~t-~~middot - ~ _ -_
-IUch PVCbQed end rap~13y the chlorlna peak the
i~~~-~~-~V~~~-t18 ~~~~ ~~ Th8
ral~~i~ JlrQP(lrUo1sof ch 1Il-ntal paek~ll1d l~t_ Q~7 -gt- ~ _~ -- gt ) gt -_ -lt ~~ the 0 -end _ xprtsntabullTIwsa changes are -r1sed ~t~ lt~ _ ~ 1nT~ IThe8llia$lb npresentthe chengelI 1n~t1V8 p~lons
ti~~~ [ ~gt 3 - - - _ - _ -
Qf nellllllllt within We surface layars of the pe1ntmiddot ru The
~f~~thtJn -d ~rfeli avr the 336 h11cxI ampScanbe
SII4iIl r~ fig 3 al~ tht rta of~ dirfers tJetwaeo thaa ltmiddot~1- 1ft~- ~ _
IJOl appfiai-ed to ~1nlt1elly until ftar day 168 it fall
~~l~ 1h8relllllrelgn1fi~ diff~be~ qrOtactlid and
proieCted (at -966 ) piIIllnts tor lioo COPPer but not folimiddottin
Ibi~tlons Of the Pain -urface 8hoWad _Pil ~ witilt1M
(Plete I to IV)~ 1hePllat1na (~) pelnt(Phta x and b)
Clearl~st-i~~r alots on the SurfllC8 IiII1kh~ter1sed ~- f hlt1 lriitUl tin conCantrat1on~ lllariy slotsre Partially
~OIIe~ ~ tile _tr~tedal middot1lere rs ~an~ densely packed
grerltr~ ~th 1ar~(iIa1niy cotifIIr) 8ntt -ampniBil (lIB1rily 1rori)being
middot vl~l~~~~~ Dtt~i~~~t~ r~ ~f~~ I~t seawatllr for 11 n IIIOre slots appeared and th~ ~~ess _trb cwer
--if -- V~ c~~ f(i ~~~i~
(Plate Ie to It)~ the granules dld not protrude from the~rtace in ~ ~~~~j1~r~-ffJmiddot0~~
suhap~~r Both 0 nd --960 at ~~pa1nts ~ - fr ~~~~-w1-U~ ~~t
sl~llrat thi stag~ (Plates Ic-d II1d xrnspac~IlYh middotmiddotWith middot middotaJ~cmiddot gt incraallO ~alolre t~ difrrqcbullbull middotbatIiI8IImiddot the protected fjd
bull ~ ~ ~t~
IIllrotacbldpaint ~~lIfIt~ftitr~ h alQt~ p~t bull u bullbullbull J ~~i~~ 1~~
lIIith IIIuper lIdguon u 4lrotatad piIJnt (Pl8ti1 II) 1hD1II8S
Ill )1IMdw1lillS gnnJla~on f-r~~ dpert1cl ~ - - - ~~r ~t
Onthe 966 1IIpetnt t-var tIIO typeaofan with diffannt c t
~hIId~(Plate UI a to c) One hIId tIII-1 bull bullbull e _ bull
fUll wJ1hleas abvlaus alats the o1fwl bed C18UU1IlOt ~litUi middotC c ~ ~
less ~ t11 h Gnnulatlan __ but ~lIiIZly def1nlld in
middot~ latter Afte fila bed tiIIrtIId 10 ~are ~~j(
extansll(tlly ~ the~tIId(O at) tIIIIple lIIIdngthe~ - ~
middot orarallet1ai leas claaI (Plate I~) f u II1ao CQ1tJAJ11d to ~Gn - -
the -966 at 1 (Plata III d tor but 1JI8V8n 1nittNdcneabull -
Althatqa -middotslots wrs abvlaus becaI1ng 1 cwampr aiI- the orwWat1an8
f1nailyftar 14 day exposure ~ fUll drMIlapIIIent on all la - ~
are extensive with the 0 lhawlng both alota pore ~
OVIKgrOloln (Plata II c to f) Gnrulatlon 1bl to ~ gt~t~~ i
or the th1cknats of tho ma ~the ~ at ~ (~~~~ themiddot - - _ middott ~ ~ ~ - -1 1 t
middot f11bull IIIIIS IIOre extans1 thIIIn at dIlya but IIIIIIIY bull1Q~ tI st11l ~w -middotr~ 1gt ~1 ~
obvious fIIany ~ loI8re still p~ yen~~~~~~~r
than before Poiu appearedto be f~lf~ila~~c-~~~a (Plate IVf~ label 1) IIIIl1ch broke~i(2)~ ~~~~
t t
bull ~lS (3) ~ rflC~~ ~i~t~~l~~KI~ t~ldiIiiicyul~loIiih ~~Rht ~ )~- -~ - ~middotd 40 ( gt ~gt ~d lt1 t ~ Jt ~remiddot4amp tha paint trix cavity bottom eroded away reVe81~ng ore
t ~~t~ por~~ ~l~ P~]Y p~~8dby di~lutiaA~ ~~~~ti~Jr~gt~ta(l~~Siinll1cl1Ud~f iUiIiI reriatlGllbullt
bull a~~ ba~tid colCriAtloriccic~ GIl ~~~d ri -966 - ib~~~amp catQn1iti~~t~I~~(~t an ( ~
t the ~~~palnts (PIa_taiiI d~~h ~wh ~ n td have ~ CamplluJMitiIVc) ~~middottti o~lgtnalrf tlt of hIght
~ l=~A ~J~rW tYtles~ p~fiiilriQcOCCold to~Ja to jU-t ~jmiddoti~ai dlatClilS ~ir -jlati ~tbullbullent aNit ~ t ~~ th8middotf~ld~ies~~t~on~ ~ -966 prQ~~~(P14ta Iii~)Omiddotiliof~m1hiiHAiiipieat ere~ll-(piitir~ p-iot~iilans(PiA-iII~ end c) nbullbulltiledt dl~by1i~~ end wirinali1sheii ~cMI~bY thI tili
bull In t-~~ ~1i1nt 1oIIIeHfd~ ~m1IS (15OClind a t ri~r-dlatoa ~lor P~t dlataiOlonlt1anGIl ttw-966
t
bull ~
pr~t~ ~is~ih iion obitJauIJ A ~rot -=1 ~ ~ i __
reprasenttild on the 966 painted 1iUrfc1ncluicunljj ~ rwt
Achnanths Colonisation by bacteria 1n the laboretory bsed
bull eliperlMnt tollMct sbiUar pattern to the fild ~
bull bull Dilicueii
bull bull DUt 1Iidnt ~lona Mve dlttllrint ~~ ictian
It ~ JntM l1tenture ttt Iff yen __ JIll the
bull ~ bicins ani 111 ~t INchlnil (PhillIPs 1t73t EWene~ 1981
In ci1ntlHt tJWY ~tit ~Wl1c iIuCh as 1rbiltyl tirI
~ lite wAlII]ltiClr dlsP8tfu~ middotthiirsttilll~intaJn~ a
bull ~
lIIifoflll ampalCentraUon through the filII 10 thoIt biocide rel_ at tha
paint surfce 1fI$Jlts in IIIOre ditfusing ~s to repla~lj -iflJ~it
resulting 111 bullbulltNdy lr leechlng rata M1llat this IIIIY be middottruIi ot
tin ~ it was nOt true ~ ttw middotlBlf fOZWJlatiGn ua ~ bull - i - bull 1(1
ObSfvt1onundr SE clarly ~ tM t1l1 to be present
~ly lIS OIIIdle-llk parUc1~~t tn lUlfiice (Ito
IV) bUt usa ~ the dIpth attt fU-middot ~y~
If one considra the concn~rtlonmiddot chMge 111 1n ~111c COKICIl8rlts fro- the paint tU- i bullbullbull tMWei b~ ~ Si6t arid
tI-~t iron oxidt 1~ can bt (Fl_ 3 IrId TIibl8I) ~t ~ ase cona1derllble d1fftrenC8s ~ a Un decz bcdYMr ttw p1iod IIIIwraS lron~- an unt Ught inCr h ~ incqJper ~tlGn occUrnd _idly at ttw ~~ Gfb ~ period ~ out ttwn fu nPldly ampQII1n ~ umiddotirid II phast This IIIOUld bt tIlq)8Ctad It cantact l-=h1ng we occun1ng
IIltfIOS8d pertlcl of copper would lMch rapldly on t-n1on 1n - shy JIsewtr There would thlll1 ba bull J-a a1iltUy leu avalUba
i1 ~ ~
_tedal becfM uposed Thi tIOUld ttwn IIllCh Thla pulSing efttct
IIIOUld be expctad to -oth OIItlllith U- putic1 __ -lllible II tot 1~ GIl a on r-x _lao Thla teet 01 put1c1e 1tUng
can tit GIl the lIUrlace ClIlC8 it ~ a pan 1amp t
a 11aM bet in Plat I to IV IIaIrIg _ lit la_ II ~ 0- ~s _- PIate IW $llldl ~bull nshyot poRI flllNltttn Al~ tin ~ lou rete thtn I bullbullbull atill Ivwnce of bull al~t pulsed 1Itue rrlict tII1IIWd
elqlCtaG inIl ttwauUr ~ I I I
0
~ i
~ ~
~I J
bullbullt bullbullbullbullt bull bull ~
bull bull
I
bull t
~~
The same seqiHllle of cevity follliltloii (intttecaseofthetestjjltle
slot- shaped) fClliOileciby SecOlld8rYtlolesfotlllingin thll cavity
bottollis releasing further deep8rtox1ils (Plate XV d and g) As
cavities_re ProdICed ~ tiecaaIelnvadedby_ter~ onelilauld
expect 1ncreaa 1n1IOdha and dIladne lMls t the t1met
which the toxlM uw ~ Ths fCMld tCtHi eo (Flvmiddot 3)
~rttno the vi that_tat ~t1on otthe~ ~lY
il1Cl1l8S8S Idth UIiJ
Differences f~in toxin li~~to Ini 966 ill 8811P~ F ~ bOth Un end ~ lcMIr levels afillW
IIIIIlfI ptaInt(~1~~1ntha~~ ~~f~~ cst rklld with tbiIccpper ElJIc~l ta~Uona invalvinv
w pl8cId at a~ht~~middot3lO~lth Un cqJpel bull
(-960)10 ttW ~af this expei1tIIri1ts thIsiorelfJltdt -
that cqlp8r b8IiIId ~ IIiIOUldProc-ln f41ip1dly t thellllgllitiv
patentW _lied () as 1t 11 further f~1t ~ witt Un
III11ch is u-tat aqulllbd Idth
it If ttda 18 eo~ ~
~ )
frOll1 paintscould bl anqnced by an illprd current clithodic
protecUansysteipos1bly ahart_nino te 11fe of the paInts
antifouling ability Bacterial coloni Uon IIIU rtdUCed on the -s66 paint This
could have been due to unflSllOUlllble electrostatic cqrge on th surtece Th1a effect CIIl pley en IIIportant role in the t~of
bacteria to alrraces (lIIarsMll19E1O) As bacteria appear to IhiJw
greater toxicltyto copper (DMpsey 1981) it is Klre likely that
enhanced cqJper leacll1nl On the -966 AIIIPl c8used the recluCed
activity The IDlol8r toxicity ofUn to bacteria (as reported by
~ -
DelIIPsey Hle1gt or its lowes leaching rat was sUpPorted bythl
relatively rapid recolonisation of the slots by lIIicroorganiSlllS ~ftar thE inlt1al aurge of leachi~ Thailllpressecl current dto ~~ the oppoSite _ff~middot ~r on dtta colonstJon l1lI-1- in the
JIbcJrtory~ta larger rUDIIra f~ fit pro~td ~ther than uriprotected Pafnta
Ccnlus1one
1 tiild-a 18 presaIIUd ~ thattJn~ ~~
~llNChng notd1rf~ ~
2 TIleuseofCP~ ~CUIftnt(at) on1h1a ~fca41ng ~- ~
pa1I]t~ to ~_lAIIach1ngn_ at toxiN - -
3 leIJstaPidbectar1al ca1on1aatJon acaampmId an ~bId Pt1ntJ ~
4 JItore larger alcrofauJing (Oiat_ and PzOtoaoana) Gr~_
occurred on the protcbld~
I l
I
bull bull ~
mfEflpoundaS middot~-t9i _jgt- ~ _ _ ~sev M~J~ (1981) (cilonJzatiooof alUf~lng lnts by _rine
Jbull ~teda9g~1ta IIarina XXlV~ 185-191
- - gt 0 Ie 1 _ - _
poundV8I$i I~lf (1~P~h 1II8rin~Ugae and Foul lng AReviw ith
t p~l-i9Jrlr ~iiMdilto5hiP-fQUllfCiH 8ot~ ~r~ ~~~ XXIVbull lIPwl~J11A
Fotteth CN~R ~kk8nGB IWPIR1seJCiiiroaic1lihd ~e ~JIIth CII Offshore St1ucturu ElllS ~tjI~f~1Dt19i4
- - ~
UKC (1976) I~races ill ~W Ecology KIuverd Univbullbull middotraabullf~iddge ~bull C~~jII illt)Ia(t RC (1980) totinllsa Cattod1CPrOtect1cinbull middotmiddot8AtEmiddot~6haOCIlfif~ T~ p No 108 ~ 18 ~ 1IIHf bull
t bull lt ~J~~~ __ ~
Norkr4irulB(198D) Advanculn UcrOfj~lpound~ltiOyVol 4 (AlItxlaquoIder fl Ed) PlerIw Press lcxIdon~ lIP~ 51-85 ~ l~l - bull bull
PJijllt~i At(fm)~mr~~JnI~1fqul1nQ JlalntlJ Riumh ~JnOrpnic toatJngs 2 192middot
)~J gt0 c ~lt -- ~ __ RogerS~ Ut (1968) ina ~(lnt~fbItliP4ili GI COnoslon ~a Technology)bull bull
Sl~iorr~ VP~Ranald~ Rctd~~ (1S80)_ ~ Experllattal Stud1aiRelate middotEffect or Cathod1c Protict1Oi withmiddot terta1nr1c
Coating Syst~ 12th Annualofr T8Id1na1ogy Co1fItHnC8 liauston TIIlt p 9-12
j
fIG1 sPct FromMon IT8TJCuZOI Int Lliu EOAX
FlG~ Fro TlTCu20Ar-4 Dy Field Ellposult I~ir
il
I c
R hi -00amp=7
-I I I I I I I I I f
iii
bull bull
bullbullbull
I J
J
I Figur 3 Thl chan In reltl rqpoIQnlof -ariOtimiddotjOmponents of the paint film with time Thl vtic~1 ampkiS unl till _n ofmiddotmt Illment per unit OJ fe 50all Cl Na
Cu StI Feeshy
PlistinllFe S7~J6 704 3~74 908
f
r e ~ ee~
middotie ~lt - ~ shybullt bull ~
t -~ l Cu~ o ~ f
t --- 3igt - bull 0 J Sn
cc
t ~
~
bullo
~ bull e
bullbullbullbull bull ~
bull ebull a QI 1- bull 0 40 431282 316
101 dmllIII)bullt bullbullbullt
hr 0 ~~bulla 5~16 ~~ -~~ i166 4S i83 8~1B
24tu 6~480 SD~83
111middot 1~1 OS6
~ 5114 6~69 -51 UIamp middot016- lt ~rmiddot
1d o Mea 818 21 021
~Z21 199 1~1Qmiddot ~Jgt2 058
14 d o ~~ 804 Q~B8~~~a~~t~middot 685
866 372amp 8amp4 OS _i~11[ ~~64
Tttile 1 A81UvlL pzQJJortlcw 0 vxl~oI ~~bull Jn 1I1Dtitctel lt-1166) end lqIlDteCt6 (O)middot~af~_~middot_lodEDf~ middotmiddotmiddotmiddotmiddotmiddotmiddotmiddotmiddotc
~ ~
_----I~
J
I bull
I
J bull tmiddot
7o CONQRESOIMTERNACIONAL
I D~CORRO$IOr(IAAINAEmiddot~USTACIONES~v c)
t 7thflfERHATlONAL CONGRESS
ON IARINE CORROSION AND fO(JUNG
~CONGRE$INtERNATIONAL DE~CORR~ON ~E ET QES~SAUSSURE$
i t
~ ~
bullt
t t
UNIVERSIDAD PourtcNlCAVollncio7J1 NiwiemIif~ 1988
(SPAN
SECcl6lfn
BiOIogro marino Morine biologY ~1i1orine
Theunpub1ishedJllBIlu8criptEl have been reprod~d asr~ielved from the
authors The SCientiUc COlIIIBittee take not respoi~ulbll~~Y for any shy
error or omssi9n
Laa ponenciasno han side publicadas y estanreproducidas tal y como
sellan recibido demiddot BUSmiddot autorea pound1 Conte Cientfico middotno se responsemiddot
bil1za de cualquier error ~ om1si~n
bullbull _ bullbull_ bullbull ___ bull $_ _ L _ - _ --- -__
SESSION II Marine biology
Friday 11th November
SURFACE PROPERTIES ON NONmiddotTOXIC ANTIFOULING PAINT FILM
K Nanishi M Murase Y Yonehara M Kishihara and T Hirama
MICROFOULING ON ALUMINIUM PANELS PLACED IN THE TROPICAL WATERS
N B Bhosle K Venkat and A B Wagh
EFFECT OF TRIBUTYLTIN OXIDE (TBTO) ON THE OVARIAN MATURATION OF THE PRAWN CARIDINA WEBERI
R Nagabhushanam B Indira R Sarojini and Mary Frances Thompson
EFFECT OF BIOACTIVE MATERIALS FROM SOFT CORALS ON BACTERIA ASSOCIATED WITH COMMON FOULER
Vitalina Mary Avelin Mary R Sarojini and R Nagabhushanam
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
SESSION I Corrosion Marine
Friday 11th November
SURFACE MODIFICATION OF CuZn20A12 ALLOY WITH PIRIMOINE DERIVATIVES
Krzysztof Debrowiecki and Kazimierz Boron
MICROENVIRONMENTAL CHANGES AROUND FOULING ORGANISMS AND THEIR CORROSION IMPLICATIONS
E G Bellinger and P Wall
THE EFFECTS OF CATHODIC PROTECTION ON THE MICROFLORA AND PERFORMANCE OF ANTIFOULING PAINTS
E G Bellinger and A J Mitchell
is usually
inhibitors
compounds
an
against
their
of
surtace
to
waters (F)
the IIOst
in
160 -190
001
The specimens Iliad ttle tOnt ot ring~ cut out from recrysshy
tallized coamercial tubes (di~ter90160 am height 200 am)
The grav~metr1c corrosiontest8 were IIIOde using an artificial
see water at room temPerature with contineous aeration
In the preUlafnatY serie8 ~t experiments lIIOClitications were
carried out by p laCirig thespeciiDens into aqueous solutions of the
chemicals specified allltWe and kept at 40middotCtor 2 hrs The
concentration 10-shy -oledJlill 111 the Ibainexperiment the
lIIOClitication conditions were follows
- Inhibitor conce~trat10n 10- to 05middot10-a ole~
-TelllPerature20~060eOmiddotC
- Time 361624 hrs The corrosion resi8tance was measured bY expoeing the samples
to aerated see water for 5 days in the preliminary experiments
or tor 5102040 or 60 daye in the main experiments
Resu1 ts and msc_sion
Only the compounds BTA DAP and BD were found in the
preliminelry experiments to have the ability ot inhibiting brass
corrosion
Tabie 2 The effectiveness of CuZn20Al2alloy surface modification after 5 day exposit 1011 to see )later
COIIIPOIolM
~
bullbullbullbullbullt
bullt
bullbullbullbullbullbullbullbullbullbullbullbullbull
naamptot~rowieCkl nd Kszilllierz BOron
Cul~AL2AiLO ~~bullocJl Inst hit 0 lnorllanic Tchnolotfy Tchntcal UnLHIrSL ty
tiL s-gttucho_JuIIO 25 60-372 IorocCaw PoLand
Introducton
The chemiCal modification ot copper alloy surtaces
carried out with compounds which are good corrosion
The most commonly used inhibitors are benzotriazole and
ot the structure similar to that ot this compound 11 As tollowsfrom a number ot reports 12-4 benzotriazole and
the compounds similar to it are chemisorbed 011 the surface of
alloy to form a stable inert layer protecting 1t
corrosion The surtace modification of copper alloys improves
stability against outdoor corrosion It also reduces the rate
corrosion in see waters In this work SOllMt other chemicals were tested as
modifiers tor CuZn20Al2 alloy (special brass) The aim was
illlProve rsistance ot the brass against corrosion in see The tollowing compounds were selected 110-phenanthroline
13-benzodiozole (BDL 2-aminopirimidine (AP) 2-mercapto-46-dishy
aminopirimidine IDAPI and 2-mercapto-456-triaminopirimidine
(TAP) hnzotriazole HITA) was also used as a reference standard
In the pniliminary experiments DAP +as fOUnd to be
effective corrosion inhibitor This compound was than studied
detail
Ettectiveness ~ Procegtdurs
BTA 293The experimenu were carried out with CuZn20Al2 brass ot the DD 12chemical compoition presented in Table 1 (accordinq to Pol ish
DAP 645Standard PH 67H-870251
1 2
I
I
~ Deteiled experi~hts were iiIodeltwi~h DAP~he most effectiv~ corrosion inhibitor among thecoiaPoUndsstudiEiq The ett~cts ot
Ii its concentration antJ~mpe~ature in modifiction bath on the bull bullbullbullbull T _
Icorrosion rate of brass in an artificial see watet afe shown in Figs bull 1-3 Fig4 pres~ht8 the) effectir~ness of middotQlqMficationvs time of exposure for the saRIPls mOdified Wlth101-M DAP at 20middotC
- bullN tor 24 brs lt
I The gra1Dtetr1~ hur~~t~dtOJCuZlj20AJ2 middotmiddotallQY in the
stationary cQnditfonaevealed that tbecorroliion rate insee water was rdUcabi~bil~ttrtr~a~~ngtl1e brass ~ur1acbullbullmiddottdtil
DAP sQlut1onTlteettbullct otmiddotcconctfrClt-tonofthe DAFIIIOltl1fYing
I solutio~ upon th~ ~~~01l10it~~t~u ~oterY ~l~~At~O)DAP 11 protetedln~iMb~n usedirttbe concentntion 10~ M The
corrosiOnitf~~it~~~a~~~~~middot tji-~iticat10n tibullbee longer aM 10nger1beDAP$oludoMmiddot of concentrations O~5middot10lt
10or~0 tUled at~oejtb~Pdht ~ott~ct ~r increuedthe
J- corrOiti(gt~ ratoflt~OAI~ lllOY~ At t~etu-ra~dtolO~Cr tl14it oclif1cationwlth ui M
DAP swtaot1by i44U~edth~ corrOiJionzte Jbi ntwsmiddotre
lusindelIndent ~t the tioCII041f1ca1iCgtl themocl1iicatlonI ffectivbullnesswasin~bisc~C(~Jtib as ~a ebull Anlncreasein modification tempra~u~ middothllda~sitive effect oli corr~ionate also for D~SolUti~ri otconciitirationoSmiddot10-X bull
For DAP of concentrations 10 and 10 Il1O ledJdI bull nof sufficient iDlProvnt in resistance ~f the brass agAinSt
corroslon tas obee~e4even at elevated temperatures 060bull orr eomiddotc The rate of corrosion 4ecreasedyithincreasing JDOdification
I time and after the 24 hrs treatment it was nearly the slUDe as
that tor uIIIIOC1it iedbi-ass For the DPA S~lutionofconcentr~~ion10-middot the change in
bull modification temperature inthe range 60-80C did not produce
significant differeneilin corr~ionratesThe resultilUJ highly reduced rate of corrosion UCQGlpQred wi thufmodified slllllPles
J remained UnChangedindep~ndentl~ of modification time The lowest rote of corrosion was obtained for the slUDplea
I I
3
modified with DPAaolutionof concentration 0510-amp at 60middotC The protection effectiveness of 97 wae obtained after a 24 hr treatment Further increase of the solution temperature to aomiddotc gaveno ItIOre improveJpent
eonc1QSJons
nllt ~u1t presente~ ~ an be S~~S~l ~n the following conclusions
1 FrOID UIOng 11 compounds stucUed the Hst corros1on
protectioopro~ruH h~ 2-mercapt0-46-timinoPii1D1idiM (DAP) 2 The protection properties of DAF 1rovtd with increasing
rilodi t1cadonterature
3 DAP wu found to tie better corraaion inhibitor tor CuZri20A12 alloy than benzotrlazole
The lIOdit~cat1on effectiveness did not cbengewitllUme of exposition ot 8amP1es to an artificial see water
Ret-nc_
1 SpinellLP~middotMaj~hM BrunonoG Trabanel1LG r~t u
KDrro 31 919-923 (1980)
2CignaRG~oG BrCOZrobullbull J bullbull US 223-225 (1979) 3 WalkerR Corratoft 2Q 290-296 (1973)
4 Trabanell1G Zuchh1F bull BrunonoG Carru1t1V t Korrobullbullbull 24 602-606 (1973)
4
-- -II
JII
if ~-it CKpirons 101- dltf1lresect
it
it ~
coyio~rOIl tit Ctn20A i ii I uy if bH 5 dys ot
exposure to ltir tltl Cl d 1 see water ~ff~CdtioH condit 10IlsI)II1
it 10-middotmoled1W te_ 2o4Q6080middot
fi92 Therote of corrosion of CuZn20Al~ 01 aff~r 5 days ofit ex~EJuHl~ortif ictal 1-eet wateYKocfificiltrorP conditIons DAPit
10-middot ~ole~ temp 20406080middotC it Fi~~TIu(~~t ~f ~orro~ionmiddot~f cuiP20A2~111i) otter 5 days of it ~~~~I~iii~~rtIti~i~l s water Moaiflcafiorl ~ohifitians DAP
~ 4 ( 1- -~~ ) ~~ II10-~ IIioledIII temp 20406080 C I ~
Fi~4 The lIlQCl~t ica~iol) effec~iven~s Yt exposure time torIt CuZn20Al2 alloy in ortiflci~t slaquolelfaterbull KoclificaqQnc~pditions
bullI it
~ Dllt~i~lQmiddot~Jel~ telllP 2PmiddotC~l hr~
ibull -
bullbull bullt
bullJ
bull bullt
bullbull 5
bullt
~ i ~
12bull 5 p
3 6
r1g 1
t bull 2OC iIrC to-c tIOC
-24
16 iMtiMtillle Ihl
I I
~ N
t ~ 5bull
JOC bull AOc bull IiOC -we I
I I I
3 1amp l I modifiCation tiMe Ihl
I I
t
f ~
_--
yshy-~~
-1shy~~(~
Jl -
- _JIJ 11 _ bull ~ bullbull L IIIJIIM
I bullbullbull shy
-
if Ilt
JII l3$i]J ~e-l~ ~ 1
J ill KItt
Ii I 1 8 Ij j iJ)Sj
emiddot Sllll0 r B 8Je Do Ji
t ~l u Ji ]
1 0
j
j
r 1l3iiiJail1h
bull III
~ i J ii i )1 t~I ~
~
~ )38 ttl i UN ~I3 11 I ~iii bullbull I fi~fi IelI M 6 6 bull s plj i I Jjhi
~l -u II11 i~ Ji ~aJ j~ ~ tll
- bull
-------------- -- - --- -- - -- shy
2
This paper desr1blS the use of ti1c~lctrooes to measure pHbull bull _ bullbull ~- l bull bull
iJQ2 chmgtS ~~ 1II1C~olones ~middotilIlg~ ~f~~s i~rsed ll ~ter ~ wlJlch t1)U11ncjotgaI11sllls are grolog SOllie p~tnt1al 11P1~~tions C1f~se cIIatlges to corrosloO at the
lt I )
surlP alllO br1lJfl)dl~ - 1 - ( _ ~ -
ItIiterWs 8nd ~ ~ ~
() poundI~~lt - -
Ttl lec~_ 1n theu bpelsnts hIM bnndeScdbed In
~~~I )iel~~bull~~i~) but n 11 ~ haJe
- Aj ~-
(1) pHel~
ThapHalec~ lII8rII basadQIJ tne-iWof It1tQ (1__)
wre the pHlt1V8recenton of~~l~ 1a fiI)OS8d bull 1Ihart po1nt extlndlngfro- tt lnsulatlnQ (Flljj1) The
pH andmiddot alUlllno~nlcate glau 1cropipettesued lnthe - - ~
electrode constnction went ~ using bull wrt1cal alectrode
puller assntielly bull vart~c81 version of the equiplllltnt
orlQlnally developed by Al8Xllllder and NasWlaquo1953) The ~ ~ -
electrolyte u-d1llllS 1N tel and filllnQ IIIIIS 8aoIpllshad using bull ~ - or
aydnge 8nd a vary fine draIIIn out Qlass capillary
Once f1lled the elfCttocJes were soaked for severalday~ in 1111
tel to allow coqlletehYdroi~s ofthe pH glass lin lnternd refelflf1(e of sllver lrecoated wi til sliver chlorld~~~s
ccinmctedmiddot to a gold plated pin iealed into the imd of the
electrode lII1th~ill~oo rubber COMPleted ei~trodei er~ stated Up downIIIards lna dilute solution of ~r~iIcrcj In allmiddot uper~ middot-temal reference Qf bull~~l
allCtradilllllS UIIIId DurlnQ -~ts -n bulllact~damiddot was c~cted to ind~ ~~F~Y(_ itrgtbull The cStcu1t layautforbotitthiiptl lIlIdpDz electtaifes
~
1a IIhIWi Sn Figure 2 bull
~ -~ - ~ -- - - - I -
CaltbnUariof all alacbodn id Clirr1ed middotOutmiddotbllfGn ftc AIIICPUIIIri inmiddot c- elctmdeiimiddotmiddotmiddot_middot~
ckalng en apars-nt by in nJIalt~ orthey dldriat
pEOCia ~lbblIt ~lntal callbraUon In ~
CIIIIi8S all ~ datafl(WtMI middotmiddotdl
(11) 0ItygIn a1acUocfes bull
TIUI catheter typa elachode use a fiat ended and of duw 1 to 15 _ It conslatad ofa ~ uia P~tJft- wlra
aeelad 1n 9l~ and IIIDadded in 1IPaKY~1o 1I1th a aUler wue KUng a raferance (see FiQ3) Ihen the epOxy hId~
h I of h l~middot careMJy ~ flat to UpQM
bath the plau _the ~lwr 1IIIIIbnre WIis ttn8llPlled
ecrosa ~ flat lind by rep8atedly d1fIPlng Jt into a aQ1utJDn Of
collodion 10 atheta1cohol A Pola1-1slng voltage of m all __
applied bit eachelectrodl and these re not thariuSed for
axper1lllental purposes until middotthe response I118sstllblallihen
bullp
4
bullit tuffer elut1on a~ a eorustllllt
it i)
~~ ~ ~eted IItld accepted for uSe elliCtlodes
it lt gt ~ ~~J ~J _ bull~ - - -gt bull
IIIIiIr~~Jn~IIOWt1on of 02 KCl D~ygen Ilectrodeser _iliry~ ~~)~ 5middot -1 ~ J~- - f -
it cli~rated aga1pst solution I of known oxygen content (as 1 w f - w ~-l _ bull bull
i IllllllaUred bY n of the l111nkler titration thad APHA 1915) ~Lmiddotmiddotlmiddot ~ ~
it Any el-=trodI not lIlCtIib1ting a ilnaar response rejected ~ ~ ~ ( _ - ~
zelia axyg8n lution obteined by the usa of a buffer
i aturated wUhpyrogallol crubbed nitrogen The 100$
~~~ reed1ng obtained by bubbling a1r thlOUltl the ~ ~~bull ~ ~ bullIII burfal far 12 hours at ~~ture 8M pressure- ~t bull
(b) [)cpars-ttal~
All -rs-tts IIIIIl1I Wried aut in atellperature controlled
bulli it
tank pliad an a teflain NYler ted l~ steel patfon ill insidil a ane _tre CIbI Faraday CIlIIIi(fig 4) Tbe~
prav1decI IIf1th a lJrfQII hinged door at the fiant for eccns and[f as gro~nded to earth to scrbullbullnout radio frequencyi intelfuwce The _tar flowing ttmluitI the experluntali - tri pullPed at a controlled rate fro a temperature
~ controlled reservoir For all experillanta involving algae
i llllA1nat1on provided using daylight fluorescent twes if l1d teel and 1Ial1ns plywood plate approxilaately 20 bull x 15
wire Ixposed in the -rine environmantfar several IIIQIIths to bull
allow a heal) growth of IIIICrofoul1ng organi_ to deVelop The
bullI steel plates had been placed utldally and IIIItre exclu8ively
bullt
~~==1$ and ~klOOrganl_ ~ldirig Itltena ~srar-nt of their
OWl The wooden plates 1IIIIIl8~sn the Jntett1ifalteclJQii
-f llllllre COIIered lIIIith ta 01 tt- alp InWfCld1IIPtth
occa1a1onal bryozaens There IIIIIIl8 IWIo lagII quantitibullbull of
~ retained -gat the algal fu-nt nPiate lIIIIIle alsO cleansed and stalJil1aad_ bllton ~
provided On 12haUrly~ ~dllrUght luar~w1 RddtUOlllll steel plat8s ~~In Alii ~dIxk~ lit
Uwrpaol (lie) IIIhare they IIIIIIl8 co~ by middot~Of~
(xtllus edulis) and the lISCld1M Clona intUn1l us
~1IIIIIl8 roved f the pJaqa NP~1id In cl_
ta tor 5 days at cansqnt ~ toIitDUampe
pH WId pOz ~1ane frc the extuMl t~~
the fouling to the Milstrata llllllre ncarded tt elecUadM
middotheld in a rigid pttllpex ~rr1et attechld to II ~ly
Jointed GIl on the lI1crcIIanipulAtor To nciad the muM the
tnraugh the foulJng tI1IIIIerds the ~ta JI_ta 0( 1 _
could be acCUllitaly controlled by talng the tIM clrlw of
the n1pJlator Iftn each lllc~ the DVIII ~oIdlft9 shy
allowed to _tabIU for ane wte prior to ncordJng the
VGltge~
~
bull a
-lt 1
l~to the_ batnacl~ 1ls~ COC)trol - fmiddotmiddotmiddot
recod4l9sof W pH 8I)d pOi at the SIJIIEdepth outside tOe 5hell middot-middotvmiddotmiddotmiddot_I~ lgt~middot middotmiddot~_middotmiddoti 1gt )
wete made
-flesolts ~
(a) pH IIIldIDz p~lllllS
The _re~ts for Hancl p02proflle tluough both algal -d - ~ gt Comiddotmiddot i -
eniIIIalfou11ng are gi~ in Figs Sto8bull --
TjpicalProtUeis daIiilttuougha t of Entronlorpha are ~1n -bull l J ltl ~~ -
Fig 5 - lIhllsttyp1cal d1innal fluctuations CICCUIJ8d during bull ~i bull bull e bull bull
photosynthetiC- II1d non-phOtinwnthet1c lods (Fig Sa) mad _~ d q _ _ __ _ i bull
decreases inpHlIIIIl1I~ IIIIa) fro-theaul-f1lCe of ttWt bullbull bull ~ bull
dcyenI tqwards the abstratacn tdch it enclOSlild (fig 511)
~fal1s1n pH--nted to as-ach as 2 units Over 9 IIIj
dIpth
figure Sa ahoid the thangea in pH -d Pl2 between 11ve barnacles
with depthln a stlat1c envirornent A steady decrease in ~
occurred as the plata IlUrfe~es toiereappro8ched pHWtially
fell on one profile before rislngbut lOSe steadily with daPth
on the second praflle SlIdlar changes are IIt10wI in fig 6b
with depth into the shells of dead barnacles 80th oxygen -d
pH levels were re~at1l1E1ly steadY ~ similar to the 8IIIblent - ~- lt
envlrOlWOt Until the -actual dead shell IIias entered When a
IIIUked drop in both fJ02 and pH CICQ11Tedbull
~llell in rigs rit JllellllUrellt$IIere maltle ~
iOlfillidual IlUs$els withtiRle (Fig 7dlb) In thll first
experisent elactrodes were placed indicated (fig)
arCUld indivicUllalsln genU flawJog ~ter The
resUlts show bull ~tle fIwrease ~ IID2 eMIS bull aiJc hour 1Ucj
The pH~t of the~ II4ttIn ~~atmiddot~
81 (te UlbientpH being 80) but 1GII8 ateIIfIU ~ the
1rtIa1ent aJphon to a6bull 111 iIadIIr ~~thI aff~ of
flOiW 00 local ~ciIndltJans~ the ~
~tadbut tile flow at~ turned Off tar 25i1aur
Jadin thelllddleofthlt ~The nailts g1R
in FlQIb -PD2 IIMtls mae ~y inflQldng CCnd1tJanamiddotbut
fell -zkadly fter alhoit lao w flaws caadawJnit a
~ fall lit 90 alii Levels rapidly ratond onca t IIIIUtchadDli agaJil No ILICh teed varlauon ~
in pH around the eldlalent 1pbn 1iIIflImI a taIdyJncn4IM -
obsmt8d aspec1elly cleM to thIIen1Ml CondltLni around
indivOJa1 ~ lIIhllst of interest are lMa repl_tative
of field conditions fro around groups or clIIIPS The
IlICpIIr1ants were ~ ~ted Wittdn denSe clUIIPs of ala
fouling a plate These r8SIJlts are given 10 figs tie (1Il1Ud
IlUlIS8ls) -d 8b (within byssal thrtiIds) In tatic ccinditJans
amprCUld the _15 u-a1(8$ ~ fellrapldly (45l) reelJ pft
rose sl1rjltly and then feU aftar 2 h llhen flowing condltiors
were resuraed after J hthe ~ rose to the original middotlevels shy
The pH rose lArlcedly at the eltictrodenear to the xhalent
bull bull bull bull bull
bull11
bullbullbullbull
it
I
8
s~ (3) but reriialned stlaadyahd ultilllateiy fell silghtiy (2)
inttie genetal fooling inassbull
~
III1thihthe1JyMli~~ Or~selci~5 IiIfieretilltti($trltus teniis ltO -=IiUlteliter~ ~ ~(Flg 8b) ~ lIIithrrolaw~ cCincentreuoos diCIppea framp1~D lBigtif3S
~bUt toaitnpl~lylt4i 150 IIifIo lIIhfjnficw t8st0ted~middot pH
t wi~1rithi thread ClUIps~lMl1BtO 74 (3) IinIt B~ito
785(2) lri-staUc cinfit1at IoihUst ettn88dgtiofthlt ~
it -flnedf81r1y~at aroOOd17S~ lIIlanf1Dlilwu
rellitoteilpft inltleUyJose (by 04 UI1ts at Ii1ta3) but it theo
bull fall only to startrfs1riq tgi1nattM not theIlXper1ilent~middot
bull Dlseusslont f tMresulblmiddot obta1nIid it ~be cl8lirly sean that f(IIJUngI
bull species vethe abUlty to Gtly IIOdlfy thdr loclmiddotmiddot
bull InIl~ts
bullbull L1t1t end dark period tluctuat1ona in algalts IBre obsarved
end ware lri close wpeeIIIen~ with those reported by Terry end
bull ~
Edyv- (1981) with 1ncnasaln both pH end IID2 llIrJngllttt
traquor1ods lIhilst thlshappened ttM surface of the t pre
bull fU_ts ware frelyprojecting into UJe water -deep intottie
mat nearer to the solid _stratlli whl1lre shading fll1lli light andbull accuIIJlatlonmiddot of deconIposJng ~gallIIBtllrial DCCIINed pH valuesbull fell IIIBrkedly Concurrent wlth this dlQp in pH ilaS II fell inbull
bullbull
9
oxygen Such t~ral and $PiItial va11ations in ~andpft
would certainly qive 11se to corrosion cells middotatmiddot the sUbsttatie
surface ~ly resulting in iorroaien pitting
The r] _tabellc ctivlUee or fau)Jng -=raJrwerttmlbullbull
together with their prapeN1ty to-=ci-1tritla~
tlwlsalves wIll also lIied to 1dcIoenv~ lIOdifktIona
Resplretory activities wU1 tend to ~~ tn-the tft and Input caapanants auch a cinia~nY qatk
1mIertabJatesmiddot ara mlk 1bullbullbull middot the bulk of their nlUagllnoua
wastes ~ excrated as ~ 1heexpftunt ncord~ pDz
end pH cningS amN 1nd1v1Q8l~ ~ ~ri
claady 1nd1cat41 the ~ of _tft~t GIl tt ~ of lnIirollliental changes Inmiddot expr_~ta lllhere no Wlter
--nts occurradthmi a np1d dacreUfi1ll pDz
COIJIdalo be a ~decreae in pH (no a) v fladrlg
condlt1C1ns are r-ed tIwn Ieymiddotmiddotor iIay nat be a 1bIIeIqI1int PH
1nctease dBpwIding ~ the --- ofmiddot iheltr ~ by the
organl1n the cluIIp 0wIgn wui a influenced by the
proxJa1ty of 1nhalent or ~a~ the dlnsity IIId the
tIlJcknes$ of the clullp w111alsa t)IMt wiuence in ~
wetr -N_tbull Hencein thick clampllP aUt and detritus
cCUlllulatlon is are COIIIIiIn ~ theh billa IIIdfbls
restricted Br8IJIIltdow1 gf organtc datrital _ter1als CI3IIIIUIIS
oxygen resulting in recLlcing conii1Uons llleasurlllenta haw
shown that sulphids qulckly build up In such deposits and
sulphate recLlcing bacteria y tie-~ SRBconcentrtlonmiddotin
J -I
II II IJI Ii ill
II
-bull I
11
10
the depOSlts woUld terid to Ile lOW at We start of ~r1ments as
condit1onswere nat favourable (POstgate 1979) The cantlooQUs
prockIctiPn of r1th f~plieUdofeacal_te1alor IlUlpllonill1l
~ts bYpoundilt8~ (~~Iick1ngand 1IkIod1) hOweva ~ t H 1 (
IIIOUld rep1dly altAIr theli11l~t and the rua~ofSRB re
notedtofncreasemiddotttcllll ( 1 SRaIll1taat llaUt 1 x 101 SMIIl-1
aftet bull -th
IUctotJlIIl corrosion canfe=t _ta1sln a1Ullberof Ways
(Iverson1972 fUlllIt end King 1975f 1Uilr 1981) pH can
aff~ the ectivity of the 1IIin organlasinuolved the H 1IIho
hava tin qlu pH range beban 10 12 (Graff 1981)
although tIdS -can be lnf1u8nCed by ather fllltors In atI1tl11n
they lIKIJlre a catbon end nitrogen aurCa 11 various
salts tin alactron accaptor (HedJart ~ idbullbull 1985)bullAil of
U- facton are availabll in the debris -alltting b41tween
ar1na foullng -d the 1lUbstra~ 110 providing bull potantlally
corrosiveanvirorwent
Evan IIIhete such accullIUlatiori does not take place varlable
changas inoxYQ8ll and pH ~trations acrossthe suface of
teel soch as C3I becaustd by foul~ ~ C3I give ti_
to corms1an calla lalildlng to piftlng corrosion
There is an initial tllld8llCY for the pH to rise as a relUlt of
the excretoryactivi1i~ of fouling aniIIal and this 1amp UIIllkely
to give rise to corros1an problems Eventually ~er feacal
h
1
~I
materials and detrltus~ill build up around or~anisms ~re bo~
low pH and p02 devul9ls providing a potentially coJJOampiIu
envinx-nt flaw does dify thh _build UfJ but inmiddotthbull
Ixparlunts carrllldotttid notprlvl(lt it R~~r it
l8Iilated the depth of deposit ~1~~ in t- ~ It ahoutd alao r~~- in f~ population so-1QIJvlWela 111111 be dying or dUd Thlb
~lng~wUl~1Ielacal1Md conoa1ve ~iccandltiorJs-t SR8a 111111 b4I fCllllfC314lng ~y pOIi1desa hlbitt~ HtInd 1tt~ WIi~ul create 1~1Mct Jrcx-ntal dlffarenca ~ ~th l1kel1haOd or pitting
n
References
Albilnder J~T 1 bL~1953~~(9EthII prockIctiPn of IdcroaltJodIIs UIIId in I~ studies Rev Sc1 lnstra ~ 528-531
_r1can Ptbl1c Health laSoc1ation 1975 5taIIdIrd~for the lIIIIIlystS of _tars end taatms 11lth pounddIt1on 1IPHIi-~ 1015 ~taenthSt NW 1IIa-l~~OC 2IJ]J6 USA
Bau-8laquo=k1ng C~IIIG amp 1IIoOd EJF1955 Biologlcal ~ in theestuarlna envi~t 1 EcolClil) of the ~ cycle Kcnlridlkjl Ned Akad -t ~ Ser B~ fi 160-181
cter DB amp SUver IA 1961 In IRafaranctlElectrodl$l EdltolS Iva DlG and l-z Gl lc8dilll1c Pr London Maw Yodlt Chapter 11 ~
Davies PIII I 8r1ric middotF 1942JII1c1oel4iitrodas for-aln local oxygen tension in n11 tissues Rlv Sci InStruII Yo 524-533
EdyileariRJGJ ~ Tetty LA UJ83 The 1nfluenc ~lIlcro algae on corrosion ofstructural steels used 4l~ the 1tIirth
Sea 81odeterioratian sect 336-347
bull bull
~
bull - -- ~It Jf bull
- ~ 13
12
Terry LA I~Ed)VeangtRfiJ 1981 lIi(toal~IOI1CGrmianF~ l)i 1St1oJhemailnEifooiiiigffixed gftshoreIt Bot 1Iar ~ 117-183 lsect~~jlaUoQ1l1 _ ~t Ef)e~gy Offs~r1I Te~lOtjjy Paj)8r
-middotltcntpI)~it$t~l91-16PP~middotmiddotdl ( I
bullbull
iIIalk e1 JL 1971 bull Ion specfie Uquid I~~ e)(charig~ ndcroelactroclels Anal ChIIaI ~8S-Q2 AGraf1~ w~S 9S1~ 1orrosloh InFIntroooctI00 tooffshOle
51~weSoes~~fab~lcat~onIns~allat1on Gulf Wall PI aelUnger EG 1982 The appl1~tlon 9fMl1ikIngmiddotHoust(jfW~Q23g-258middot icroelectrade in the study of pH end PD2 in lIHinli
HerbierC SN fGH~rt~ )RI -Stoctcdalpii Watltif1Sal RS ~1ronIIInb Ub~~ 27l-Z79 1985 Fac~or lontro1l11)9 tha activity ofmiddot sulpMte
bull Wall P 1983 The appllcatlon Md ~ ofiwrJ)e~c~odl shy-taicfii9 tiiCtetili tn~ diiilncj tar infection
techniques for the iiaurnt cif icraeov1ror1liltital5QiCiety ofJebo1- Engi~ SPE 139781-10 _i~~ - ~ ~ i ( _ IKIdif1catlons caued by aGII8 rlnefou~lng -C~ebullbull
PhD lhaaia Ibiverslty or ~tar UKbull bullbull bull -
IIH1nke JA 19fiSa In EXperllRants in Physiology and middotampioClIiIiiilatJift l c E(If(orltGAiKetkutgt ~c-Presa-
Wooltdngton AObullbull Davenport J 1983 pH lind IlIl2 IMla ffl~- j~t~5 -lMIIleath 1IH1na ro-fauUngGqWl J bull Expbull 1
B101 Earlsectsect 113-124 Klnke JAIII 1969b II) Glass IUcroelectJQdes poundx1tors Uwal1~Ui~ OrOllt arid~t Nt JOhn fII11ey YOI1lt thaptar 11
~-f _f~ ~ ~~nt ~ Igt
bull Iverson IIIP 1972 Biolog~ Qrroaian In Advencesin
corrosion scianca and tecIYIology 2EdS FCntaia G~ I bull IIS~et RIIIbull p~ PJaSsLancIoIl 1-42
bull
Jorgensen BB Reusbech NP I Cohen T 19B3 Photosynthesis and strUcture of bIIn~ II1crobWp MC1OIIlectrocki and ~ stud1ea of four cyanabacledalt -~usllWtOl ~bullbull 1~f093
t l~ O~ ~~~ H~ ~9s7~c-gtHatste~iecfn1k van shy pellediftlrtenPt - Stlchelektraden (1-Su
AiJBitiKlificfiliilislflilr)zlIr polrapusdiailJIIissui1g_ liIserstoff(jruQkas fur d1eB lIsUbullbullung der IIliJlttOUf1cillatlon PflftgaHArdI ~fl39
bullbull Mller JOA 1981 tals In EconcinI1c II1croblolooy sect McNbUdbicldatariOtationEd Rosebull A middotAcadelc Praulondcn149-202
t Mller J DAI King RA 1975 8io$~iorat1on of lllltals In lIIi~1al ClflPlCts of the deterloration of tarWbullbull EdsGilbertRJ~ middotloveloCk 0111 bullAcadeic- Presst IILandon 8Jo103
bullt
IPotgate JR 1976 The sulphate-reducingbacteda
Callbridge tIn1versity Press Cliilbrldg8U-K
Reusbecb fUI i Ward DIII 1964 i~~~1Ida5tud1el of
bullt inter$Utilll IIIItez dttm1stry and photosynthetk activity
in ilflaI spring Idcrob1altAppl~EIi~ MerobW tsect4IlU-27lt I
IIbullt bullt
bullbull Ibullt
1
Inmiddot middotmiddot11middotmiddot f~~ a~ -tmiddot bull J~f
I~ W
i5iImiddot J~a t
ilmiddotmiddotmiddot I middotIt Ii
l
~
middotmiddot~~middotlL= g r
Lj Ic
t
i ~
I
gi II cEi ~ I I
1 t rW
D
tR bull
bull t 10
11
r
i
I ~
j
~
I
~bullbullbullbullbullbullbullbullbullbullbull
LV
FigS pH and p02 profiles into macrofoltling eomlllllllitilS dominated by Balm trenatU$ on steel plate 11 two pH profiles Ib) two lIOai profiles leI pH and P02 profiles into dead barllllCle shiella withdetri~Us I I
itIbl r p02 ~ I
nvnHg 11 70
j ifJi eo 1amp0
laquof it 30 s
I 20 N
ca 11 10 a o -- shy i bull
11
80
19
781 o 2 6 8 10 12 0 2 6 amiddot 10 12 dIpdI Qf ~iftto fouling In HighIst numbIII pIatIi
8deg~~
~i c5 iioo5 fiil
pH 8 bull_ ~
~~ ~
_ bullbullbull- pH
-pOz
(cJ
e-Clf ( j~JJlaquo1- --~I ~a ilil - _ I i ]1
~ ri en i ~ oJt ~iJ id tUI
2u1U I shyI t
shy1 2 3 6 6 7 8 9 10 11 12 i shyDepth 9f electrode (mm) into fouling
1 i
1
~----------~-------
~
~-~ lt1~
middotmiddotPmiddot ~
I
V
~
ill
1 Tbellitt~~ of cathodiC protection OO~~lIic~tlO aridrfOflllUlc
ofi AA~Uoullr14l ~1~I
I
ael~lner EG middotIUtcb8l1AJ Dept lt4 lInY1roMe1lal Bl01011
=~~venilt7 IlariCheW 1f139PL $ ~11al1df
r
I
Ablitrict ~~~ e~l billttI ~ted with ~pert1n ant11_1111ampp paint llere itpoii~1n _vatI- In the tl~ld iUIit 1bOioatbrt~ScM Were
1IUbjll~ to qtbocJ~C ~teltt1on byen _ current (~)SJi inci EDAX Iieamp~ta IJdlctecI ~ lachln teoc copper OIl tbePlCtec~ pa1n~ atter14 dQ1a~ protected Pllnt abo ebowed
) reduCed bacteiial activIty butmiddot ~e4 dltoe Iu1d protOzoanmiddot co~~l~tlont~1l IlU l~byenCOCltact not byen dUfwIl00 ~r _baRl lrillolvedln tOXln nil ereio noted
l ~~
1
I
Introduction
Applications of protective coatings and cathodic protect1on(cp~l
are recognised corrosion control methods for IIIBrine steel Itructureso Cathod~c protectlOll preventl conCIsion by Int~lng til8ctrl~l
currentS trem externalllClUfCa to e~~tthlJno~ e~ carrollon feetlans C04U lUChu tUauJ1ng Aelnts a
(
barrIer to ~ the flao of couos1a1 ~aboIHnu~r
Dccurrlng anodes cathodes or wlwit gelvWc CCIIII1 ( 19681
flinger 1980) AnUfauling ~ also JJICOqIOrII~ b1oc~~
Tr~ylt1n nuor~ (~) end o~ Oxide (~) (~ flJ1J
- of u blacld1111~
V _Ut Evans 1981) ~ ~
end growth of bipfoullng 0I1I)IIIII1IIIIe Wdd~ ca ~ of
paint rusendmiddotmanemiddot ~(r~ til )
As corrosian ~t1on ~ bICCIIIII -an ~1tlQd ~ of
antiroullnQs 1 conjunction _ith CP have bllClIOr ~
resultIng in reseudt 1rwestllj111tlng trw COIIIPIitlbUlty or VU~ ~t t _~
rOratlation to dlfrert levI ot CP (~Eelf 18110 ~
~t 1980) RelatIvely 1nIestiljlllUans have ~ urtw ~
or CP an thIt anUraullngpert~ of palnta
In thIt preseot 1nlIestioaUon tV~iln111 ronutlon ud
cOIIIHrclallY incanjlntlon wIth CP was ~t4IC~ Iw I sraed current (-966 l1li) arid exposed at the 1 Strait IInQlbullbullbully
~t1ons weremiddot 00 the IIhcIrt bnifects the PprIUd_
had an a) the various COIIIPQI1eIIts of n pe1ntf1l11 end b) l1oft~
developaentbull
bull bull ~
~ ~ - f
bull ~ _T~~-middot~ ~~- ~r _ -$1
(1) ~rlttionor Painted Steel Billets middot_ ~~tmiddot_tlJ middot~hmiddot~--~fmiddot ~ middot~A~-~~middot - -~ ~
Thirty-al ateel blllets (8-10aD2 end l depth) were cut frollbullt ~middot~middot~middotmiddot~middottrSmiddott ~jS-~ It bull gt~ tfl - middot--imiddotmiddot ~ t1dItrd steal plates The dimenSlonsof -thB butits-lim chosento
bull -~~ middotmiddotgt2h ~ ~-Imiddot ~~rt ~- -middoti~~ ni ~~middot1 l middot-~t~~middot _
(a) fecilltate amiddot Ie detrucUve -thxI of prePraUonpdor to ~~_gt gt~ k7i ~_ t~ 1--1- _ ) - +--_~ -_~bull ming electron ~ 4II1CI (iialloWdirectobs8iVatlCnS of the
- i lt-- ~ gtbull -o1e billet 8Uface
gt r~middotmiddotj ~L(~middot~middot rmiddot tmiddot J ~ - ~ - lt _ bull Single non-standard (1hIula~) nickel lIIireSmiddot Spot Welded e1Cing
bull ~1deJ ~ b~i~ ~j~~ tri afl~1b1ipOi~ f ~1t1 ~4middotmiddot f-gt -~ ~S-- ( -- - - _bull resin EactI billet then Undb~ tI1d prtiladlllltha5O __ eyilr
bull ~~_ ~~imiddot ~1iil h~ intlfcili~ ~eonbt1nlyi Tr1butylt1n F~id(18n)~~~O~dettu20i~blcicld8is 1IIfii bull i - ~ ( then 1IPI)l1ed to glvaadry Ulil tIi1c1ltnOss or 1(1)-150bullbull
tbull (2) IipreSSeCi Current Syat
bull A rigid ~tic due 1I8S drWedllliihwrtv-s41 liiIihOieS IIPProX~
bullbull 05-10 cia episrt ~ Pilnted ampUl8t Wsfbed IntaPQSIUon ampy
threadlng the nickel IIIlr8throu11 the dinto a tarproot junction
box Eliltilen of th8~1l1ii (recelving ~t-d curnritlere IIIlred
Into bull block connectOrrecelvlng dc trOll is potentiOstat The
rainJng iontroi blUets Wet8fbedfl11i1iiy lIIithln thejtlytlonbox
i~ityd~c IkiliPly ~~ ~lat1ruaWiode 8ncIacalOliel
refer~ electtodetoiere fllied ~ the per1aetat of the plut1cdlc IIIId
connected to the pot8ritlostat The desl~ of the potentlosbt ailoiolEld
~PlDteItlO1TClJrnnt to be MIllntained at 8prampset levtl of 966 II1II
c
r~
Grid raf 1J5s-t 114iH 563724 dud 0 Juna
Alolecule andbechr1a in general are ~ to
sutface lI1eroleyer (nlUSton) of _ter (~bullbull 191KJ)~To avold
contaJnatlonof ttw painted surf by U- -vents
blllets Were s-nact in a 1 conte1ner or atc1le (02i ) f~~nd
ter Iih1ch then f9ICI in tv 1IIhere the b~
be r-oved undeNeter El~~ bllleta fa Mcb trtai bullbullel
removed over a 1 dtq (336 hr~ expoeure perioda
each being ~~bn uposurnof 1 5 11 iA 168and DlIIauz1r
Arter field axpauHli the billets wan
fUtered _tar to ~ 1_ debtlflMd far 10 aJnuta ~
wv ~~~endattid sdbUiUd(otO
~ ~l f~ nn1ng el~~ --
lOOted onto 1middotCIi ~bullbullWIsend carbonmiddotcoa~nmiddotmiddot~middot f
each billet Ws~ Iis1ng a ccr1dge ~el~~
(5360) IIIlth anergy dJ8perslve X-ray alcroenelre1a (mx)
_lysis the les Wen reNed end coated III1th vold to al]Ql En
detailed sat of the alaquofa1l
(4) Lfiborattux ~
Painted taa~ bUletalllllre pnparad ~
in a Mrlne reclrculet1ng flOlll _ ~
f1Oll the ~ ~a1t~y Tha~~Un bullybullbullbull
10C M1nta1ned ~~t1as of ~t~la protazOllbull~bull10shy
(Entermap) ~ ~~~middotNlt~Eamp1~ __~and ~ r - ~ -
certaJn ~lc 1IQ1~ 1(1 the
the painted
~
thre repl1catft fn
plKed 1nto(022_)
- thIine1J~~ -lt bull
Atter mAX
(1-2) and i-n-t bn collected
14OC
Pain~
-
12 daY~ exposure and prepared as in d)~ - t
~I ~~~-- i~yen 1~ v
Rflltsi ~ _ ~f-~lI ft j bull _
rt~ll0~~~llY~~~~peln~stetsl~11i~ex~sed for
y~t1~J~ ~~~~1~1~~1UII~ya~kooJ~~~lt ~ ~~_~~I1J~ ~~~tomAX -lysis ~era than gold ~ bull bullbullbull c - _ bull lt
~~fllr ~~~ vllUll ~t1ans of the rfee htures and gt Co
~f~ - ~~~ ~f ~ ~aobta1Mdue IjIll811 iI1Flg 1~2 The IIIIdn ~~ ~ lt)ji9 )tmiddotmiddot - ht
- bull
~I rIIplt~ tIt8 -~ cOlllponanta of the pei1t 1bullbull WelIIlItr1x $ilj~t-~~middot - ~ _ -_
-IUch PVCbQed end rap~13y the chlorlna peak the
i~~~-~~-~V~~~-t18 ~~~~ ~~ Th8
ral~~i~ JlrQP(lrUo1sof ch 1Il-ntal paek~ll1d l~t_ Q~7 -gt- ~ _~ -- gt ) gt -_ -lt ~~ the 0 -end _ xprtsntabullTIwsa changes are -r1sed ~t~ lt~ _ ~ 1nT~ IThe8llia$lb npresentthe chengelI 1n~t1V8 p~lons
ti~~~ [ ~gt 3 - - - _ - _ -
Qf nellllllllt within We surface layars of the pe1ntmiddot ru The
~f~~thtJn -d ~rfeli avr the 336 h11cxI ampScanbe
SII4iIl r~ fig 3 al~ tht rta of~ dirfers tJetwaeo thaa ltmiddot~1- 1ft~- ~ _
IJOl appfiai-ed to ~1nlt1elly until ftar day 168 it fall
~~l~ 1h8relllllrelgn1fi~ diff~be~ qrOtactlid and
proieCted (at -966 ) piIIllnts tor lioo COPPer but not folimiddottin
Ibi~tlons Of the Pain -urface 8hoWad _Pil ~ witilt1M
(Plete I to IV)~ 1hePllat1na (~) pelnt(Phta x and b)
Clearl~st-i~~r alots on the SurfllC8 IiII1kh~ter1sed ~- f hlt1 lriitUl tin conCantrat1on~ lllariy slotsre Partially
~OIIe~ ~ tile _tr~tedal middot1lere rs ~an~ densely packed
grerltr~ ~th 1ar~(iIa1niy cotifIIr) 8ntt -ampniBil (lIB1rily 1rori)being
middot vl~l~~~~~ Dtt~i~~~t~ r~ ~f~~ I~t seawatllr for 11 n IIIOre slots appeared and th~ ~~ess _trb cwer
--if -- V~ c~~ f(i ~~~i~
(Plate Ie to It)~ the granules dld not protrude from the~rtace in ~ ~~~~j1~r~-ffJmiddot0~~
suhap~~r Both 0 nd --960 at ~~pa1nts ~ - fr ~~~~-w1-U~ ~~t
sl~llrat thi stag~ (Plates Ic-d II1d xrnspac~IlYh middotmiddotWith middot middotaJ~cmiddot gt incraallO ~alolre t~ difrrqcbullbull middotbatIiI8IImiddot the protected fjd
bull ~ ~ ~t~
IIllrotacbldpaint ~~lIfIt~ftitr~ h alQt~ p~t bull u bullbullbull J ~~i~~ 1~~
lIIith IIIuper lIdguon u 4lrotatad piIJnt (Pl8ti1 II) 1hD1II8S
Ill )1IMdw1lillS gnnJla~on f-r~~ dpert1cl ~ - - - ~~r ~t
Onthe 966 1IIpetnt t-var tIIO typeaofan with diffannt c t
~hIId~(Plate UI a to c) One hIId tIII-1 bull bullbull e _ bull
fUll wJ1hleas abvlaus alats the o1fwl bed C18UU1IlOt ~litUi middotC c ~ ~
less ~ t11 h Gnnulatlan __ but ~lIiIZly def1nlld in
middot~ latter Afte fila bed tiIIrtIId 10 ~are ~~j(
extansll(tlly ~ the~tIId(O at) tIIIIple lIIIdngthe~ - ~
middot orarallet1ai leas claaI (Plate I~) f u II1ao CQ1tJAJ11d to ~Gn - -
the -966 at 1 (Plata III d tor but 1JI8V8n 1nittNdcneabull -
Althatqa -middotslots wrs abvlaus becaI1ng 1 cwampr aiI- the orwWat1an8
f1nailyftar 14 day exposure ~ fUll drMIlapIIIent on all la - ~
are extensive with the 0 lhawlng both alota pore ~
OVIKgrOloln (Plata II c to f) Gnrulatlon 1bl to ~ gt~t~~ i
or the th1cknats of tho ma ~the ~ at ~ (~~~~ themiddot - - _ middott ~ ~ ~ - -1 1 t
middot f11bull IIIIIS IIOre extans1 thIIIn at dIlya but IIIIIIIY bull1Q~ tI st11l ~w -middotr~ 1gt ~1 ~
obvious fIIany ~ loI8re still p~ yen~~~~~~~r
than before Poiu appearedto be f~lf~ila~~c-~~~a (Plate IVf~ label 1) IIIIl1ch broke~i(2)~ ~~~~
t t
bull ~lS (3) ~ rflC~~ ~i~t~~l~~KI~ t~ldiIiiicyul~loIiih ~~Rht ~ )~- -~ - ~middotd 40 ( gt ~gt ~d lt1 t ~ Jt ~remiddot4amp tha paint trix cavity bottom eroded away reVe81~ng ore
t ~~t~ por~~ ~l~ P~]Y p~~8dby di~lutiaA~ ~~~~ti~Jr~gt~ta(l~~Siinll1cl1Ud~f iUiIiI reriatlGllbullt
bull a~~ ba~tid colCriAtloriccic~ GIl ~~~d ri -966 - ib~~~amp catQn1iti~~t~I~~(~t an ( ~
t the ~~~palnts (PIa_taiiI d~~h ~wh ~ n td have ~ CamplluJMitiIVc) ~~middottti o~lgtnalrf tlt of hIght
~ l=~A ~J~rW tYtles~ p~fiiilriQcOCCold to~Ja to jU-t ~jmiddoti~ai dlatClilS ~ir -jlati ~tbullbullent aNit ~ t ~~ th8middotf~ld~ies~~t~on~ ~ -966 prQ~~~(P14ta Iii~)Omiddotiliof~m1hiiHAiiipieat ere~ll-(piitir~ p-iot~iilans(PiA-iII~ end c) nbullbulltiledt dl~by1i~~ end wirinali1sheii ~cMI~bY thI tili
bull In t-~~ ~1i1nt 1oIIIeHfd~ ~m1IS (15OClind a t ri~r-dlatoa ~lor P~t dlataiOlonlt1anGIl ttw-966
t
bull ~
pr~t~ ~is~ih iion obitJauIJ A ~rot -=1 ~ ~ i __
reprasenttild on the 966 painted 1iUrfc1ncluicunljj ~ rwt
Achnanths Colonisation by bacteria 1n the laboretory bsed
bull eliperlMnt tollMct sbiUar pattern to the fild ~
bull bull Dilicueii
bull bull DUt 1Iidnt ~lona Mve dlttllrint ~~ ictian
It ~ JntM l1tenture ttt Iff yen __ JIll the
bull ~ bicins ani 111 ~t INchlnil (PhillIPs 1t73t EWene~ 1981
In ci1ntlHt tJWY ~tit ~Wl1c iIuCh as 1rbiltyl tirI
~ lite wAlII]ltiClr dlsP8tfu~ middotthiirsttilll~intaJn~ a
bull ~
lIIifoflll ampalCentraUon through the filII 10 thoIt biocide rel_ at tha
paint surfce 1fI$Jlts in IIIOre ditfusing ~s to repla~lj -iflJ~it
resulting 111 bullbulltNdy lr leechlng rata M1llat this IIIIY be middottruIi ot
tin ~ it was nOt true ~ ttw middotlBlf fOZWJlatiGn ua ~ bull - i - bull 1(1
ObSfvt1onundr SE clarly ~ tM t1l1 to be present
~ly lIS OIIIdle-llk parUc1~~t tn lUlfiice (Ito
IV) bUt usa ~ the dIpth attt fU-middot ~y~
If one considra the concn~rtlonmiddot chMge 111 1n ~111c COKICIl8rlts fro- the paint tU- i bullbullbull tMWei b~ ~ Si6t arid
tI-~t iron oxidt 1~ can bt (Fl_ 3 IrId TIibl8I) ~t ~ ase cona1derllble d1fftrenC8s ~ a Un decz bcdYMr ttw p1iod IIIIwraS lron~- an unt Ught inCr h ~ incqJper ~tlGn occUrnd _idly at ttw ~~ Gfb ~ period ~ out ttwn fu nPldly ampQII1n ~ umiddotirid II phast This IIIOUld bt tIlq)8Ctad It cantact l-=h1ng we occun1ng
IIltfIOS8d pertlcl of copper would lMch rapldly on t-n1on 1n - shy JIsewtr There would thlll1 ba bull J-a a1iltUy leu avalUba
i1 ~ ~
_tedal becfM uposed Thi tIOUld ttwn IIllCh Thla pulSing efttct
IIIOUld be expctad to -oth OIItlllith U- putic1 __ -lllible II tot 1~ GIl a on r-x _lao Thla teet 01 put1c1e 1tUng
can tit GIl the lIUrlace ClIlC8 it ~ a pan 1amp t
a 11aM bet in Plat I to IV IIaIrIg _ lit la_ II ~ 0- ~s _- PIate IW $llldl ~bull nshyot poRI flllNltttn Al~ tin ~ lou rete thtn I bullbullbull atill Ivwnce of bull al~t pulsed 1Itue rrlict tII1IIWd
elqlCtaG inIl ttwauUr ~ I I I
0
~ i
~ ~
~I J
bullbullt bullbullbullbullt bull bull ~
bull bull
I
bull t
~~
The same seqiHllle of cevity follliltloii (intttecaseofthetestjjltle
slot- shaped) fClliOileciby SecOlld8rYtlolesfotlllingin thll cavity
bottollis releasing further deep8rtox1ils (Plate XV d and g) As
cavities_re ProdICed ~ tiecaaIelnvadedby_ter~ onelilauld
expect 1ncreaa 1n1IOdha and dIladne lMls t the t1met
which the toxlM uw ~ Ths fCMld tCtHi eo (Flvmiddot 3)
~rttno the vi that_tat ~t1on otthe~ ~lY
il1Cl1l8S8S Idth UIiJ
Differences f~in toxin li~~to Ini 966 ill 8811P~ F ~ bOth Un end ~ lcMIr levels afillW
IIIIIlfI ptaInt(~1~~1ntha~~ ~~f~~ cst rklld with tbiIccpper ElJIc~l ta~Uona invalvinv
w pl8cId at a~ht~~middot3lO~lth Un cqJpel bull
(-960)10 ttW ~af this expei1tIIri1ts thIsiorelfJltdt -
that cqlp8r b8IiIId ~ IIiIOUldProc-ln f41ip1dly t thellllgllitiv
patentW _lied () as 1t 11 further f~1t ~ witt Un
III11ch is u-tat aqulllbd Idth
it If ttda 18 eo~ ~
~ )
frOll1 paintscould bl anqnced by an illprd current clithodic
protecUansysteipos1bly ahart_nino te 11fe of the paInts
antifouling ability Bacterial coloni Uon IIIU rtdUCed on the -s66 paint This
could have been due to unflSllOUlllble electrostatic cqrge on th surtece Th1a effect CIIl pley en IIIportant role in the t~of
bacteria to alrraces (lIIarsMll19E1O) As bacteria appear to IhiJw
greater toxicltyto copper (DMpsey 1981) it is Klre likely that
enhanced cqJper leacll1nl On the -966 AIIIPl c8used the recluCed
activity The IDlol8r toxicity ofUn to bacteria (as reported by
~ -
DelIIPsey Hle1gt or its lowes leaching rat was sUpPorted bythl
relatively rapid recolonisation of the slots by lIIicroorganiSlllS ~ftar thE inlt1al aurge of leachi~ Thailllpressecl current dto ~~ the oppoSite _ff~middot ~r on dtta colonstJon l1lI-1- in the
JIbcJrtory~ta larger rUDIIra f~ fit pro~td ~ther than uriprotected Pafnta
Ccnlus1one
1 tiild-a 18 presaIIUd ~ thattJn~ ~~
~llNChng notd1rf~ ~
2 TIleuseofCP~ ~CUIftnt(at) on1h1a ~fca41ng ~- ~
pa1I]t~ to ~_lAIIach1ngn_ at toxiN - -
3 leIJstaPidbectar1al ca1on1aatJon acaampmId an ~bId Pt1ntJ ~
4 JItore larger alcrofauJing (Oiat_ and PzOtoaoana) Gr~_
occurred on the protcbld~
I l
I
bull bull ~
mfEflpoundaS middot~-t9i _jgt- ~ _ _ ~sev M~J~ (1981) (cilonJzatiooof alUf~lng lnts by _rine
Jbull ~teda9g~1ta IIarina XXlV~ 185-191
- - gt 0 Ie 1 _ - _
poundV8I$i I~lf (1~P~h 1II8rin~Ugae and Foul lng AReviw ith
t p~l-i9Jrlr ~iiMdilto5hiP-fQUllfCiH 8ot~ ~r~ ~~~ XXIVbull lIPwl~J11A
Fotteth CN~R ~kk8nGB IWPIR1seJCiiiroaic1lihd ~e ~JIIth CII Offshore St1ucturu ElllS ~tjI~f~1Dt19i4
- - ~
UKC (1976) I~races ill ~W Ecology KIuverd Univbullbull middotraabullf~iddge ~bull C~~jII illt)Ia(t RC (1980) totinllsa Cattod1CPrOtect1cinbull middotmiddot8AtEmiddot~6haOCIlfif~ T~ p No 108 ~ 18 ~ 1IIHf bull
t bull lt ~J~~~ __ ~
Norkr4irulB(198D) Advanculn UcrOfj~lpound~ltiOyVol 4 (AlItxlaquoIder fl Ed) PlerIw Press lcxIdon~ lIP~ 51-85 ~ l~l - bull bull
PJijllt~i At(fm)~mr~~JnI~1fqul1nQ JlalntlJ Riumh ~JnOrpnic toatJngs 2 192middot
)~J gt0 c ~lt -- ~ __ RogerS~ Ut (1968) ina ~(lnt~fbItliP4ili GI COnoslon ~a Technology)bull bull
Sl~iorr~ VP~Ranald~ Rctd~~ (1S80)_ ~ Experllattal Stud1aiRelate middotEffect or Cathod1c Protict1Oi withmiddot terta1nr1c
Coating Syst~ 12th Annualofr T8Id1na1ogy Co1fItHnC8 liauston TIIlt p 9-12
j
fIG1 sPct FromMon IT8TJCuZOI Int Lliu EOAX
FlG~ Fro TlTCu20Ar-4 Dy Field Ellposult I~ir
il
I c
R hi -00amp=7
-I I I I I I I I I f
iii
bull bull
bullbullbull
I J
J
I Figur 3 Thl chan In reltl rqpoIQnlof -ariOtimiddotjOmponents of the paint film with time Thl vtic~1 ampkiS unl till _n ofmiddotmt Illment per unit OJ fe 50all Cl Na
Cu StI Feeshy
PlistinllFe S7~J6 704 3~74 908
f
r e ~ ee~
middotie ~lt - ~ shybullt bull ~
t -~ l Cu~ o ~ f
t --- 3igt - bull 0 J Sn
cc
t ~
~
bullo
~ bull e
bullbullbullbull bull ~
bull ebull a QI 1- bull 0 40 431282 316
101 dmllIII)bullt bullbullbullt
hr 0 ~~bulla 5~16 ~~ -~~ i166 4S i83 8~1B
24tu 6~480 SD~83
111middot 1~1 OS6
~ 5114 6~69 -51 UIamp middot016- lt ~rmiddot
1d o Mea 818 21 021
~Z21 199 1~1Qmiddot ~Jgt2 058
14 d o ~~ 804 Q~B8~~~a~~t~middot 685
866 372amp 8amp4 OS _i~11[ ~~64
Tttile 1 A81UvlL pzQJJortlcw 0 vxl~oI ~~bull Jn 1I1Dtitctel lt-1166) end lqIlDteCt6 (O)middot~af~_~middot_lodEDf~ middotmiddotmiddotmiddotmiddotmiddotmiddotmiddotmiddotc
~ ~
_----I~
J
I bull
I
J bull tmiddot
7o CONQRESOIMTERNACIONAL
I D~CORRO$IOr(IAAINAEmiddot~USTACIONES~v c)
t 7thflfERHATlONAL CONGRESS
ON IARINE CORROSION AND fO(JUNG
~CONGRE$INtERNATIONAL DE~CORR~ON ~E ET QES~SAUSSURE$
i t
~ ~
bullt
t t
UNIVERSIDAD PourtcNlCAVollncio7J1 NiwiemIif~ 1988
(SPAN
SECcl6lfn
BiOIogro marino Morine biologY ~1i1orine
Theunpub1ishedJllBIlu8criptEl have been reprod~d asr~ielved from the
authors The SCientiUc COlIIIBittee take not respoi~ulbll~~Y for any shy
error or omssi9n
Laa ponenciasno han side publicadas y estanreproducidas tal y como
sellan recibido demiddot BUSmiddot autorea pound1 Conte Cientfico middotno se responsemiddot
bil1za de cualquier error ~ om1si~n
bullbull _ bullbull_ bullbull ___ bull $_ _ L _ - _ --- -__
SESSION II Marine biology
Friday 11th November
SURFACE PROPERTIES ON NONmiddotTOXIC ANTIFOULING PAINT FILM
K Nanishi M Murase Y Yonehara M Kishihara and T Hirama
MICROFOULING ON ALUMINIUM PANELS PLACED IN THE TROPICAL WATERS
N B Bhosle K Venkat and A B Wagh
EFFECT OF TRIBUTYLTIN OXIDE (TBTO) ON THE OVARIAN MATURATION OF THE PRAWN CARIDINA WEBERI
R Nagabhushanam B Indira R Sarojini and Mary Frances Thompson
EFFECT OF BIOACTIVE MATERIALS FROM SOFT CORALS ON BACTERIA ASSOCIATED WITH COMMON FOULER
Vitalina Mary Avelin Mary R Sarojini and R Nagabhushanam
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
is usually
inhibitors
compounds
an
against
their
of
surtace
to
waters (F)
the IIOst
in
160 -190
001
The specimens Iliad ttle tOnt ot ring~ cut out from recrysshy
tallized coamercial tubes (di~ter90160 am height 200 am)
The grav~metr1c corrosiontest8 were IIIOde using an artificial
see water at room temPerature with contineous aeration
In the preUlafnatY serie8 ~t experiments lIIOClitications were
carried out by p laCirig thespeciiDens into aqueous solutions of the
chemicals specified allltWe and kept at 40middotCtor 2 hrs The
concentration 10-shy -oledJlill 111 the Ibainexperiment the
lIIOClitication conditions were follows
- Inhibitor conce~trat10n 10- to 05middot10-a ole~
-TelllPerature20~060eOmiddotC
- Time 361624 hrs The corrosion resi8tance was measured bY expoeing the samples
to aerated see water for 5 days in the preliminary experiments
or tor 5102040 or 60 daye in the main experiments
Resu1 ts and msc_sion
Only the compounds BTA DAP and BD were found in the
preliminelry experiments to have the ability ot inhibiting brass
corrosion
Tabie 2 The effectiveness of CuZn20Al2alloy surface modification after 5 day exposit 1011 to see )later
COIIIPOIolM
~
bullbullbullbullbullt
bullt
bullbullbullbullbullbullbullbullbullbullbullbullbull
naamptot~rowieCkl nd Kszilllierz BOron
Cul~AL2AiLO ~~bullocJl Inst hit 0 lnorllanic Tchnolotfy Tchntcal UnLHIrSL ty
tiL s-gttucho_JuIIO 25 60-372 IorocCaw PoLand
Introducton
The chemiCal modification ot copper alloy surtaces
carried out with compounds which are good corrosion
The most commonly used inhibitors are benzotriazole and
ot the structure similar to that ot this compound 11 As tollowsfrom a number ot reports 12-4 benzotriazole and
the compounds similar to it are chemisorbed 011 the surface of
alloy to form a stable inert layer protecting 1t
corrosion The surtace modification of copper alloys improves
stability against outdoor corrosion It also reduces the rate
corrosion in see waters In this work SOllMt other chemicals were tested as
modifiers tor CuZn20Al2 alloy (special brass) The aim was
illlProve rsistance ot the brass against corrosion in see The tollowing compounds were selected 110-phenanthroline
13-benzodiozole (BDL 2-aminopirimidine (AP) 2-mercapto-46-dishy
aminopirimidine IDAPI and 2-mercapto-456-triaminopirimidine
(TAP) hnzotriazole HITA) was also used as a reference standard
In the pniliminary experiments DAP +as fOUnd to be
effective corrosion inhibitor This compound was than studied
detail
Ettectiveness ~ Procegtdurs
BTA 293The experimenu were carried out with CuZn20Al2 brass ot the DD 12chemical compoition presented in Table 1 (accordinq to Pol ish
DAP 645Standard PH 67H-870251
1 2
I
I
~ Deteiled experi~hts were iiIodeltwi~h DAP~he most effectiv~ corrosion inhibitor among thecoiaPoUndsstudiEiq The ett~cts ot
Ii its concentration antJ~mpe~ature in modifiction bath on the bull bullbullbullbull T _
Icorrosion rate of brass in an artificial see watet afe shown in Figs bull 1-3 Fig4 pres~ht8 the) effectir~ness of middotQlqMficationvs time of exposure for the saRIPls mOdified Wlth101-M DAP at 20middotC
- bullN tor 24 brs lt
I The gra1Dtetr1~ hur~~t~dtOJCuZlj20AJ2 middotmiddotallQY in the
stationary cQnditfonaevealed that tbecorroliion rate insee water was rdUcabi~bil~ttrtr~a~~ngtl1e brass ~ur1acbullbullmiddottdtil
DAP sQlut1onTlteettbullct otmiddotcconctfrClt-tonofthe DAFIIIOltl1fYing
I solutio~ upon th~ ~~~01l10it~~t~u ~oterY ~l~~At~O)DAP 11 protetedln~iMb~n usedirttbe concentntion 10~ M The
corrosiOnitf~~it~~~a~~~~~middot tji-~iticat10n tibullbee longer aM 10nger1beDAP$oludoMmiddot of concentrations O~5middot10lt
10or~0 tUled at~oejtb~Pdht ~ott~ct ~r increuedthe
J- corrOiti(gt~ ratoflt~OAI~ lllOY~ At t~etu-ra~dtolO~Cr tl14it oclif1cationwlth ui M
DAP swtaot1by i44U~edth~ corrOiJionzte Jbi ntwsmiddotre
lusindelIndent ~t the tioCII041f1ca1iCgtl themocl1iicatlonI ffectivbullnesswasin~bisc~C(~Jtib as ~a ebull Anlncreasein modification tempra~u~ middothllda~sitive effect oli corr~ionate also for D~SolUti~ri otconciitirationoSmiddot10-X bull
For DAP of concentrations 10 and 10 Il1O ledJdI bull nof sufficient iDlProvnt in resistance ~f the brass agAinSt
corroslon tas obee~e4even at elevated temperatures 060bull orr eomiddotc The rate of corrosion 4ecreasedyithincreasing JDOdification
I time and after the 24 hrs treatment it was nearly the slUDe as
that tor uIIIIOC1it iedbi-ass For the DPA S~lutionofconcentr~~ion10-middot the change in
bull modification temperature inthe range 60-80C did not produce
significant differeneilin corr~ionratesThe resultilUJ highly reduced rate of corrosion UCQGlpQred wi thufmodified slllllPles
J remained UnChangedindep~ndentl~ of modification time The lowest rote of corrosion was obtained for the slUDplea
I I
3
modified with DPAaolutionof concentration 0510-amp at 60middotC The protection effectiveness of 97 wae obtained after a 24 hr treatment Further increase of the solution temperature to aomiddotc gaveno ItIOre improveJpent
eonc1QSJons
nllt ~u1t presente~ ~ an be S~~S~l ~n the following conclusions
1 FrOID UIOng 11 compounds stucUed the Hst corros1on
protectioopro~ruH h~ 2-mercapt0-46-timinoPii1D1idiM (DAP) 2 The protection properties of DAF 1rovtd with increasing
rilodi t1cadonterature
3 DAP wu found to tie better corraaion inhibitor tor CuZri20A12 alloy than benzotrlazole
The lIOdit~cat1on effectiveness did not cbengewitllUme of exposition ot 8amP1es to an artificial see water
Ret-nc_
1 SpinellLP~middotMaj~hM BrunonoG Trabanel1LG r~t u
KDrro 31 919-923 (1980)
2CignaRG~oG BrCOZrobullbull J bullbull US 223-225 (1979) 3 WalkerR Corratoft 2Q 290-296 (1973)
4 Trabanell1G Zuchh1F bull BrunonoG Carru1t1V t Korrobullbullbull 24 602-606 (1973)
4
-- -II
JII
if ~-it CKpirons 101- dltf1lresect
it
it ~
coyio~rOIl tit Ctn20A i ii I uy if bH 5 dys ot
exposure to ltir tltl Cl d 1 see water ~ff~CdtioH condit 10IlsI)II1
it 10-middotmoled1W te_ 2o4Q6080middot
fi92 Therote of corrosion of CuZn20Al~ 01 aff~r 5 days ofit ex~EJuHl~ortif ictal 1-eet wateYKocfificiltrorP conditIons DAPit
10-middot ~ole~ temp 20406080middotC it Fi~~TIu(~~t ~f ~orro~ionmiddot~f cuiP20A2~111i) otter 5 days of it ~~~~I~iii~~rtIti~i~l s water Moaiflcafiorl ~ohifitians DAP
~ 4 ( 1- -~~ ) ~~ II10-~ IIioledIII temp 20406080 C I ~
Fi~4 The lIlQCl~t ica~iol) effec~iven~s Yt exposure time torIt CuZn20Al2 alloy in ortiflci~t slaquolelfaterbull KoclificaqQnc~pditions
bullI it
~ Dllt~i~lQmiddot~Jel~ telllP 2PmiddotC~l hr~
ibull -
bullbull bullt
bullJ
bull bullt
bullbull 5
bullt
~ i ~
12bull 5 p
3 6
r1g 1
t bull 2OC iIrC to-c tIOC
-24
16 iMtiMtillle Ihl
I I
~ N
t ~ 5bull
JOC bull AOc bull IiOC -we I
I I I
3 1amp l I modifiCation tiMe Ihl
I I
t
f ~
_--
yshy-~~
-1shy~~(~
Jl -
- _JIJ 11 _ bull ~ bullbull L IIIJIIM
I bullbullbull shy
-
if Ilt
JII l3$i]J ~e-l~ ~ 1
J ill KItt
Ii I 1 8 Ij j iJ)Sj
emiddot Sllll0 r B 8Je Do Ji
t ~l u Ji ]
1 0
j
j
r 1l3iiiJail1h
bull III
~ i J ii i )1 t~I ~
~
~ )38 ttl i UN ~I3 11 I ~iii bullbull I fi~fi IelI M 6 6 bull s plj i I Jjhi
~l -u II11 i~ Ji ~aJ j~ ~ tll
- bull
-------------- -- - --- -- - -- shy
2
This paper desr1blS the use of ti1c~lctrooes to measure pHbull bull _ bullbull ~- l bull bull
iJQ2 chmgtS ~~ 1II1C~olones ~middotilIlg~ ~f~~s i~rsed ll ~ter ~ wlJlch t1)U11ncjotgaI11sllls are grolog SOllie p~tnt1al 11P1~~tions C1f~se cIIatlges to corrosloO at the
lt I )
surlP alllO br1lJfl)dl~ - 1 - ( _ ~ -
ItIiterWs 8nd ~ ~ ~
() poundI~~lt - -
Ttl lec~_ 1n theu bpelsnts hIM bnndeScdbed In
~~~I )iel~~bull~~i~) but n 11 ~ haJe
- Aj ~-
(1) pHel~
ThapHalec~ lII8rII basadQIJ tne-iWof It1tQ (1__)
wre the pHlt1V8recenton of~~l~ 1a fiI)OS8d bull 1Ihart po1nt extlndlngfro- tt lnsulatlnQ (Flljj1) The
pH andmiddot alUlllno~nlcate glau 1cropipettesued lnthe - - ~
electrode constnction went ~ using bull wrt1cal alectrode
puller assntielly bull vart~c81 version of the equiplllltnt
orlQlnally developed by Al8Xllllder and NasWlaquo1953) The ~ ~ -
electrolyte u-d1llllS 1N tel and filllnQ IIIIIS 8aoIpllshad using bull ~ - or
aydnge 8nd a vary fine draIIIn out Qlass capillary
Once f1lled the elfCttocJes were soaked for severalday~ in 1111
tel to allow coqlletehYdroi~s ofthe pH glass lin lnternd refelflf1(e of sllver lrecoated wi til sliver chlorld~~~s
ccinmctedmiddot to a gold plated pin iealed into the imd of the
electrode lII1th~ill~oo rubber COMPleted ei~trodei er~ stated Up downIIIards lna dilute solution of ~r~iIcrcj In allmiddot uper~ middot-temal reference Qf bull~~l
allCtradilllllS UIIIId DurlnQ -~ts -n bulllact~damiddot was c~cted to ind~ ~~F~Y(_ itrgtbull The cStcu1t layautforbotitthiiptl lIlIdpDz electtaifes
~
1a IIhIWi Sn Figure 2 bull
~ -~ - ~ -- - - - I -
CaltbnUariof all alacbodn id Clirr1ed middotOutmiddotbllfGn ftc AIIICPUIIIri inmiddot c- elctmdeiimiddotmiddotmiddot_middot~
ckalng en apars-nt by in nJIalt~ orthey dldriat
pEOCia ~lbblIt ~lntal callbraUon In ~
CIIIIi8S all ~ datafl(WtMI middotmiddotdl
(11) 0ItygIn a1acUocfes bull
TIUI catheter typa elachode use a fiat ended and of duw 1 to 15 _ It conslatad ofa ~ uia P~tJft- wlra
aeelad 1n 9l~ and IIIDadded in 1IPaKY~1o 1I1th a aUler wue KUng a raferance (see FiQ3) Ihen the epOxy hId~
h I of h l~middot careMJy ~ flat to UpQM
bath the plau _the ~lwr 1IIIIIbnre WIis ttn8llPlled
ecrosa ~ flat lind by rep8atedly d1fIPlng Jt into a aQ1utJDn Of
collodion 10 atheta1cohol A Pola1-1slng voltage of m all __
applied bit eachelectrodl and these re not thariuSed for
axper1lllental purposes until middotthe response I118sstllblallihen
bullp
4
bullit tuffer elut1on a~ a eorustllllt
it i)
~~ ~ ~eted IItld accepted for uSe elliCtlodes
it lt gt ~ ~~J ~J _ bull~ - - -gt bull
IIIIiIr~~Jn~IIOWt1on of 02 KCl D~ygen Ilectrodeser _iliry~ ~~)~ 5middot -1 ~ J~- - f -
it cli~rated aga1pst solution I of known oxygen content (as 1 w f - w ~-l _ bull bull
i IllllllaUred bY n of the l111nkler titration thad APHA 1915) ~Lmiddotmiddotlmiddot ~ ~
it Any el-=trodI not lIlCtIib1ting a ilnaar response rejected ~ ~ ~ ( _ - ~
zelia axyg8n lution obteined by the usa of a buffer
i aturated wUhpyrogallol crubbed nitrogen The 100$
~~~ reed1ng obtained by bubbling a1r thlOUltl the ~ ~~bull ~ ~ bullIII burfal far 12 hours at ~~ture 8M pressure- ~t bull
(b) [)cpars-ttal~
All -rs-tts IIIIIl1I Wried aut in atellperature controlled
bulli it
tank pliad an a teflain NYler ted l~ steel patfon ill insidil a ane _tre CIbI Faraday CIlIIIi(fig 4) Tbe~
prav1decI IIf1th a lJrfQII hinged door at the fiant for eccns and[f as gro~nded to earth to scrbullbullnout radio frequencyi intelfuwce The _tar flowing ttmluitI the experluntali - tri pullPed at a controlled rate fro a temperature
~ controlled reservoir For all experillanta involving algae
i llllA1nat1on provided using daylight fluorescent twes if l1d teel and 1Ial1ns plywood plate approxilaately 20 bull x 15
wire Ixposed in the -rine environmantfar several IIIQIIths to bull
allow a heal) growth of IIIICrofoul1ng organi_ to deVelop The
bullI steel plates had been placed utldally and IIIItre exclu8ively
bullt
~~==1$ and ~klOOrganl_ ~ldirig Itltena ~srar-nt of their
OWl The wooden plates 1IIIIIl8~sn the Jntett1ifalteclJQii
-f llllllre COIIered lIIIith ta 01 tt- alp InWfCld1IIPtth
occa1a1onal bryozaens There IIIIIIl8 IWIo lagII quantitibullbull of
~ retained -gat the algal fu-nt nPiate lIIIIIle alsO cleansed and stalJil1aad_ bllton ~
provided On 12haUrly~ ~dllrUght luar~w1 RddtUOlllll steel plat8s ~~In Alii ~dIxk~ lit
Uwrpaol (lie) IIIhare they IIIIIIl8 co~ by middot~Of~
(xtllus edulis) and the lISCld1M Clona intUn1l us
~1IIIIIl8 roved f the pJaqa NP~1id In cl_
ta tor 5 days at cansqnt ~ toIitDUampe
pH WId pOz ~1ane frc the extuMl t~~
the fouling to the Milstrata llllllre ncarded tt elecUadM
middotheld in a rigid pttllpex ~rr1et attechld to II ~ly
Jointed GIl on the lI1crcIIanipulAtor To nciad the muM the
tnraugh the foulJng tI1IIIIerds the ~ta JI_ta 0( 1 _
could be acCUllitaly controlled by talng the tIM clrlw of
the n1pJlator Iftn each lllc~ the DVIII ~oIdlft9 shy
allowed to _tabIU for ane wte prior to ncordJng the
VGltge~
~
bull a
-lt 1
l~to the_ batnacl~ 1ls~ COC)trol - fmiddotmiddotmiddot
recod4l9sof W pH 8I)d pOi at the SIJIIEdepth outside tOe 5hell middot-middotvmiddotmiddotmiddot_I~ lgt~middot middotmiddot~_middotmiddoti 1gt )
wete made
-flesolts ~
(a) pH IIIldIDz p~lllllS
The _re~ts for Hancl p02proflle tluough both algal -d - ~ gt Comiddotmiddot i -
eniIIIalfou11ng are gi~ in Figs Sto8bull --
TjpicalProtUeis daIiilttuougha t of Entronlorpha are ~1n -bull l J ltl ~~ -
Fig 5 - lIhllsttyp1cal d1innal fluctuations CICCUIJ8d during bull ~i bull bull e bull bull
photosynthetiC- II1d non-phOtinwnthet1c lods (Fig Sa) mad _~ d q _ _ __ _ i bull
decreases inpHlIIIIl1I~ IIIIa) fro-theaul-f1lCe of ttWt bullbull bull ~ bull
dcyenI tqwards the abstratacn tdch it enclOSlild (fig 511)
~fal1s1n pH--nted to as-ach as 2 units Over 9 IIIj
dIpth
figure Sa ahoid the thangea in pH -d Pl2 between 11ve barnacles
with depthln a stlat1c envirornent A steady decrease in ~
occurred as the plata IlUrfe~es toiereappro8ched pHWtially
fell on one profile before rislngbut lOSe steadily with daPth
on the second praflle SlIdlar changes are IIt10wI in fig 6b
with depth into the shells of dead barnacles 80th oxygen -d
pH levels were re~at1l1E1ly steadY ~ similar to the 8IIIblent - ~- lt
envlrOlWOt Until the -actual dead shell IIias entered When a
IIIUked drop in both fJ02 and pH CICQ11Tedbull
~llell in rigs rit JllellllUrellt$IIere maltle ~
iOlfillidual IlUs$els withtiRle (Fig 7dlb) In thll first
experisent elactrodes were placed indicated (fig)
arCUld indivicUllalsln genU flawJog ~ter The
resUlts show bull ~tle fIwrease ~ IID2 eMIS bull aiJc hour 1Ucj
The pH~t of the~ II4ttIn ~~atmiddot~
81 (te UlbientpH being 80) but 1GII8 ateIIfIU ~ the
1rtIa1ent aJphon to a6bull 111 iIadIIr ~~thI aff~ of
flOiW 00 local ~ciIndltJans~ the ~
~tadbut tile flow at~ turned Off tar 25i1aur
Jadin thelllddleofthlt ~The nailts g1R
in FlQIb -PD2 IIMtls mae ~y inflQldng CCnd1tJanamiddotbut
fell -zkadly fter alhoit lao w flaws caadawJnit a
~ fall lit 90 alii Levels rapidly ratond onca t IIIIUtchadDli agaJil No ILICh teed varlauon ~
in pH around the eldlalent 1pbn 1iIIflImI a taIdyJncn4IM -
obsmt8d aspec1elly cleM to thIIen1Ml CondltLni around
indivOJa1 ~ lIIhllst of interest are lMa repl_tative
of field conditions fro around groups or clIIIPS The
IlICpIIr1ants were ~ ~ted Wittdn denSe clUIIPs of ala
fouling a plate These r8SIJlts are given 10 figs tie (1Il1Ud
IlUlIS8ls) -d 8b (within byssal thrtiIds) In tatic ccinditJans
amprCUld the _15 u-a1(8$ ~ fellrapldly (45l) reelJ pft
rose sl1rjltly and then feU aftar 2 h llhen flowing condltiors
were resuraed after J hthe ~ rose to the original middotlevels shy
The pH rose lArlcedly at the eltictrodenear to the xhalent
bull bull bull bull bull
bull11
bullbullbullbull
it
I
8
s~ (3) but reriialned stlaadyahd ultilllateiy fell silghtiy (2)
inttie genetal fooling inassbull
~
III1thihthe1JyMli~~ Or~selci~5 IiIfieretilltti($trltus teniis ltO -=IiUlteliter~ ~ ~(Flg 8b) ~ lIIithrrolaw~ cCincentreuoos diCIppea framp1~D lBigtif3S
~bUt toaitnpl~lylt4i 150 IIifIo lIIhfjnficw t8st0ted~middot pH
t wi~1rithi thread ClUIps~lMl1BtO 74 (3) IinIt B~ito
785(2) lri-staUc cinfit1at IoihUst ettn88dgtiofthlt ~
it -flnedf81r1y~at aroOOd17S~ lIIlanf1Dlilwu
rellitoteilpft inltleUyJose (by 04 UI1ts at Ii1ta3) but it theo
bull fall only to startrfs1riq tgi1nattM not theIlXper1ilent~middot
bull Dlseusslont f tMresulblmiddot obta1nIid it ~be cl8lirly sean that f(IIJUngI
bull species vethe abUlty to Gtly IIOdlfy thdr loclmiddotmiddot
bull InIl~ts
bullbull L1t1t end dark period tluctuat1ona in algalts IBre obsarved
end ware lri close wpeeIIIen~ with those reported by Terry end
bull ~
Edyv- (1981) with 1ncnasaln both pH end IID2 llIrJngllttt
traquor1ods lIhilst thlshappened ttM surface of the t pre
bull fU_ts ware frelyprojecting into UJe water -deep intottie
mat nearer to the solid _stratlli whl1lre shading fll1lli light andbull accuIIJlatlonmiddot of deconIposJng ~gallIIBtllrial DCCIINed pH valuesbull fell IIIBrkedly Concurrent wlth this dlQp in pH ilaS II fell inbull
bullbull
9
oxygen Such t~ral and $PiItial va11ations in ~andpft
would certainly qive 11se to corrosion cells middotatmiddot the sUbsttatie
surface ~ly resulting in iorroaien pitting
The r] _tabellc ctivlUee or fau)Jng -=raJrwerttmlbullbull
together with their prapeN1ty to-=ci-1tritla~
tlwlsalves wIll also lIied to 1dcIoenv~ lIOdifktIona
Resplretory activities wU1 tend to ~~ tn-the tft and Input caapanants auch a cinia~nY qatk
1mIertabJatesmiddot ara mlk 1bullbullbull middot the bulk of their nlUagllnoua
wastes ~ excrated as ~ 1heexpftunt ncord~ pDz
end pH cningS amN 1nd1v1Q8l~ ~ ~ri
claady 1nd1cat41 the ~ of _tft~t GIl tt ~ of lnIirollliental changes Inmiddot expr_~ta lllhere no Wlter
--nts occurradthmi a np1d dacreUfi1ll pDz
COIJIdalo be a ~decreae in pH (no a) v fladrlg
condlt1C1ns are r-ed tIwn Ieymiddotmiddotor iIay nat be a 1bIIeIqI1int PH
1nctease dBpwIding ~ the --- ofmiddot iheltr ~ by the
organl1n the cluIIp 0wIgn wui a influenced by the
proxJa1ty of 1nhalent or ~a~ the dlnsity IIId the
tIlJcknes$ of the clullp w111alsa t)IMt wiuence in ~
wetr -N_tbull Hencein thick clampllP aUt and detritus
cCUlllulatlon is are COIIIIiIn ~ theh billa IIIdfbls
restricted Br8IJIIltdow1 gf organtc datrital _ter1als CI3IIIIUIIS
oxygen resulting in recLlcing conii1Uons llleasurlllenta haw
shown that sulphids qulckly build up In such deposits and
sulphate recLlcing bacteria y tie-~ SRBconcentrtlonmiddotin
J -I
II II IJI Ii ill
II
-bull I
11
10
the depOSlts woUld terid to Ile lOW at We start of ~r1ments as
condit1onswere nat favourable (POstgate 1979) The cantlooQUs
prockIctiPn of r1th f~plieUdofeacal_te1alor IlUlpllonill1l
~ts bYpoundilt8~ (~~Iick1ngand 1IkIod1) hOweva ~ t H 1 (
IIIOUld rep1dly altAIr theli11l~t and the rua~ofSRB re
notedtofncreasemiddotttcllll ( 1 SRaIll1taat llaUt 1 x 101 SMIIl-1
aftet bull -th
IUctotJlIIl corrosion canfe=t _ta1sln a1Ullberof Ways
(Iverson1972 fUlllIt end King 1975f 1Uilr 1981) pH can
aff~ the ectivity of the 1IIin organlasinuolved the H 1IIho
hava tin qlu pH range beban 10 12 (Graff 1981)
although tIdS -can be lnf1u8nCed by ather fllltors In atI1tl11n
they lIKIJlre a catbon end nitrogen aurCa 11 various
salts tin alactron accaptor (HedJart ~ idbullbull 1985)bullAil of
U- facton are availabll in the debris -alltting b41tween
ar1na foullng -d the 1lUbstra~ 110 providing bull potantlally
corrosiveanvirorwent
Evan IIIhete such accullIUlatiori does not take place varlable
changas inoxYQ8ll and pH ~trations acrossthe suface of
teel soch as C3I becaustd by foul~ ~ C3I give ti_
to corms1an calla lalildlng to piftlng corrosion
There is an initial tllld8llCY for the pH to rise as a relUlt of
the excretoryactivi1i~ of fouling aniIIal and this 1amp UIIllkely
to give rise to corros1an problems Eventually ~er feacal
h
1
~I
materials and detrltus~ill build up around or~anisms ~re bo~
low pH and p02 devul9ls providing a potentially coJJOampiIu
envinx-nt flaw does dify thh _build UfJ but inmiddotthbull
Ixparlunts carrllldotttid notprlvl(lt it R~~r it
l8Iilated the depth of deposit ~1~~ in t- ~ It ahoutd alao r~~- in f~ population so-1QIJvlWela 111111 be dying or dUd Thlb
~lng~wUl~1Ielacal1Md conoa1ve ~iccandltiorJs-t SR8a 111111 b4I fCllllfC314lng ~y pOIi1desa hlbitt~ HtInd 1tt~ WIi~ul create 1~1Mct Jrcx-ntal dlffarenca ~ ~th l1kel1haOd or pitting
n
References
Albilnder J~T 1 bL~1953~~(9EthII prockIctiPn of IdcroaltJodIIs UIIId in I~ studies Rev Sc1 lnstra ~ 528-531
_r1can Ptbl1c Health laSoc1ation 1975 5taIIdIrd~for the lIIIIIlystS of _tars end taatms 11lth pounddIt1on 1IPHIi-~ 1015 ~taenthSt NW 1IIa-l~~OC 2IJ]J6 USA
Bau-8laquo=k1ng C~IIIG amp 1IIoOd EJF1955 Biologlcal ~ in theestuarlna envi~t 1 EcolClil) of the ~ cycle Kcnlridlkjl Ned Akad -t ~ Ser B~ fi 160-181
cter DB amp SUver IA 1961 In IRafaranctlElectrodl$l EdltolS Iva DlG and l-z Gl lc8dilll1c Pr London Maw Yodlt Chapter 11 ~
Davies PIII I 8r1ric middotF 1942JII1c1oel4iitrodas for-aln local oxygen tension in n11 tissues Rlv Sci InStruII Yo 524-533
EdyileariRJGJ ~ Tetty LA UJ83 The 1nfluenc ~lIlcro algae on corrosion ofstructural steels used 4l~ the 1tIirth
Sea 81odeterioratian sect 336-347
bull bull
~
bull - -- ~It Jf bull
- ~ 13
12
Terry LA I~Ed)VeangtRfiJ 1981 lIi(toal~IOI1CGrmianF~ l)i 1St1oJhemailnEifooiiiigffixed gftshoreIt Bot 1Iar ~ 117-183 lsect~~jlaUoQ1l1 _ ~t Ef)e~gy Offs~r1I Te~lOtjjy Paj)8r
-middotltcntpI)~it$t~l91-16PP~middotmiddotdl ( I
bullbull
iIIalk e1 JL 1971 bull Ion specfie Uquid I~~ e)(charig~ ndcroelactroclels Anal ChIIaI ~8S-Q2 AGraf1~ w~S 9S1~ 1orrosloh InFIntroooctI00 tooffshOle
51~weSoes~~fab~lcat~onIns~allat1on Gulf Wall PI aelUnger EG 1982 The appl1~tlon 9fMl1ikIngmiddotHoust(jfW~Q23g-258middot icroelectrade in the study of pH end PD2 in lIHinli
HerbierC SN fGH~rt~ )RI -Stoctcdalpii Watltif1Sal RS ~1ronIIInb Ub~~ 27l-Z79 1985 Fac~or lontro1l11)9 tha activity ofmiddot sulpMte
bull Wall P 1983 The appllcatlon Md ~ ofiwrJ)e~c~odl shy-taicfii9 tiiCtetili tn~ diiilncj tar infection
techniques for the iiaurnt cif icraeov1ror1liltital5QiCiety ofJebo1- Engi~ SPE 139781-10 _i~~ - ~ ~ i ( _ IKIdif1catlons caued by aGII8 rlnefou~lng -C~ebullbull
PhD lhaaia Ibiverslty or ~tar UKbull bullbull bull -
IIH1nke JA 19fiSa In EXperllRants in Physiology and middotampioClIiIiiilatJift l c E(If(orltGAiKetkutgt ~c-Presa-
Wooltdngton AObullbull Davenport J 1983 pH lind IlIl2 IMla ffl~- j~t~5 -lMIIleath 1IH1na ro-fauUngGqWl J bull Expbull 1
B101 Earlsectsect 113-124 Klnke JAIII 1969b II) Glass IUcroelectJQdes poundx1tors Uwal1~Ui~ OrOllt arid~t Nt JOhn fII11ey YOI1lt thaptar 11
~-f _f~ ~ ~~nt ~ Igt
bull Iverson IIIP 1972 Biolog~ Qrroaian In Advencesin
corrosion scianca and tecIYIology 2EdS FCntaia G~ I bull IIS~et RIIIbull p~ PJaSsLancIoIl 1-42
bull
Jorgensen BB Reusbech NP I Cohen T 19B3 Photosynthesis and strUcture of bIIn~ II1crobWp MC1OIIlectrocki and ~ stud1ea of four cyanabacledalt -~usllWtOl ~bullbull 1~f093
t l~ O~ ~~~ H~ ~9s7~c-gtHatste~iecfn1k van shy pellediftlrtenPt - Stlchelektraden (1-Su
AiJBitiKlificfiliilislflilr)zlIr polrapusdiailJIIissui1g_ liIserstoff(jruQkas fur d1eB lIsUbullbullung der IIliJlttOUf1cillatlon PflftgaHArdI ~fl39
bullbull Mller JOA 1981 tals In EconcinI1c II1croblolooy sect McNbUdbicldatariOtationEd Rosebull A middotAcadelc Praulondcn149-202
t Mller J DAI King RA 1975 8io$~iorat1on of lllltals In lIIi~1al ClflPlCts of the deterloration of tarWbullbull EdsGilbertRJ~ middotloveloCk 0111 bullAcadeic- Presst IILandon 8Jo103
bullt
IPotgate JR 1976 The sulphate-reducingbacteda
Callbridge tIn1versity Press Cliilbrldg8U-K
Reusbecb fUI i Ward DIII 1964 i~~~1Ida5tud1el of
bullt inter$Utilll IIIItez dttm1stry and photosynthetk activity
in ilflaI spring Idcrob1altAppl~EIi~ MerobW tsect4IlU-27lt I
IIbullt bullt
bullbull Ibullt
1
Inmiddot middotmiddot11middotmiddot f~~ a~ -tmiddot bull J~f
I~ W
i5iImiddot J~a t
ilmiddotmiddotmiddot I middotIt Ii
l
~
middotmiddot~~middotlL= g r
Lj Ic
t
i ~
I
gi II cEi ~ I I
1 t rW
D
tR bull
bull t 10
11
r
i
I ~
j
~
I
~bullbullbullbullbullbullbullbullbullbullbull
LV
FigS pH and p02 profiles into macrofoltling eomlllllllitilS dominated by Balm trenatU$ on steel plate 11 two pH profiles Ib) two lIOai profiles leI pH and P02 profiles into dead barllllCle shiella withdetri~Us I I
itIbl r p02 ~ I
nvnHg 11 70
j ifJi eo 1amp0
laquof it 30 s
I 20 N
ca 11 10 a o -- shy i bull
11
80
19
781 o 2 6 8 10 12 0 2 6 amiddot 10 12 dIpdI Qf ~iftto fouling In HighIst numbIII pIatIi
8deg~~
~i c5 iioo5 fiil
pH 8 bull_ ~
~~ ~
_ bullbullbull- pH
-pOz
(cJ
e-Clf ( j~JJlaquo1- --~I ~a ilil - _ I i ]1
~ ri en i ~ oJt ~iJ id tUI
2u1U I shyI t
shy1 2 3 6 6 7 8 9 10 11 12 i shyDepth 9f electrode (mm) into fouling
1 i
1
~----------~-------
~
~-~ lt1~
middotmiddotPmiddot ~
I
V
~
ill
1 Tbellitt~~ of cathodiC protection OO~~lIic~tlO aridrfOflllUlc
ofi AA~Uoullr14l ~1~I
I
ael~lner EG middotIUtcb8l1AJ Dept lt4 lInY1roMe1lal Bl01011
=~~venilt7 IlariCheW 1f139PL $ ~11al1df
r
I
Ablitrict ~~~ e~l billttI ~ted with ~pert1n ant11_1111ampp paint llere itpoii~1n _vatI- In the tl~ld iUIit 1bOioatbrt~ScM Were
1IUbjll~ to qtbocJ~C ~teltt1on byen _ current (~)SJi inci EDAX Iieamp~ta IJdlctecI ~ lachln teoc copper OIl tbePlCtec~ pa1n~ atter14 dQ1a~ protected Pllnt abo ebowed
) reduCed bacteiial activIty butmiddot ~e4 dltoe Iu1d protOzoanmiddot co~~l~tlont~1l IlU l~byenCOCltact not byen dUfwIl00 ~r _baRl lrillolvedln tOXln nil ereio noted
l ~~
1
I
Introduction
Applications of protective coatings and cathodic protect1on(cp~l
are recognised corrosion control methods for IIIBrine steel Itructureso Cathod~c protectlOll preventl conCIsion by Int~lng til8ctrl~l
currentS trem externalllClUfCa to e~~tthlJno~ e~ carrollon feetlans C04U lUChu tUauJ1ng Aelnts a
(
barrIer to ~ the flao of couos1a1 ~aboIHnu~r
Dccurrlng anodes cathodes or wlwit gelvWc CCIIII1 ( 19681
flinger 1980) AnUfauling ~ also JJICOqIOrII~ b1oc~~
Tr~ylt1n nuor~ (~) end o~ Oxide (~) (~ flJ1J
- of u blacld1111~
V _Ut Evans 1981) ~ ~
end growth of bipfoullng 0I1I)IIIII1IIIIe Wdd~ ca ~ of
paint rusendmiddotmanemiddot ~(r~ til )
As corrosian ~t1on ~ bICCIIIII -an ~1tlQd ~ of
antiroullnQs 1 conjunction _ith CP have bllClIOr ~
resultIng in reseudt 1rwestllj111tlng trw COIIIPIitlbUlty or VU~ ~t t _~
rOratlation to dlfrert levI ot CP (~Eelf 18110 ~
~t 1980) RelatIvely 1nIestiljlllUans have ~ urtw ~
or CP an thIt anUraullngpert~ of palnta
In thIt preseot 1nlIestioaUon tV~iln111 ronutlon ud
cOIIIHrclallY incanjlntlon wIth CP was ~t4IC~ Iw I sraed current (-966 l1li) arid exposed at the 1 Strait IInQlbullbullbully
~t1ons weremiddot 00 the IIhcIrt bnifects the PprIUd_
had an a) the various COIIIPQI1eIIts of n pe1ntf1l11 end b) l1oft~
developaentbull
bull bull ~
~ ~ - f
bull ~ _T~~-middot~ ~~- ~r _ -$1
(1) ~rlttionor Painted Steel Billets middot_ ~~tmiddot_tlJ middot~hmiddot~--~fmiddot ~ middot~A~-~~middot - -~ ~
Thirty-al ateel blllets (8-10aD2 end l depth) were cut frollbullt ~middot~middot~middotmiddot~middottrSmiddott ~jS-~ It bull gt~ tfl - middot--imiddotmiddot ~ t1dItrd steal plates The dimenSlonsof -thB butits-lim chosento
bull -~~ middotmiddotgt2h ~ ~-Imiddot ~~rt ~- -middoti~~ ni ~~middot1 l middot-~t~~middot _
(a) fecilltate amiddot Ie detrucUve -thxI of prePraUonpdor to ~~_gt gt~ k7i ~_ t~ 1--1- _ ) - +--_~ -_~bull ming electron ~ 4II1CI (iialloWdirectobs8iVatlCnS of the
- i lt-- ~ gtbull -o1e billet 8Uface
gt r~middotmiddotj ~L(~middot~middot rmiddot tmiddot J ~ - ~ - lt _ bull Single non-standard (1hIula~) nickel lIIireSmiddot Spot Welded e1Cing
bull ~1deJ ~ b~i~ ~j~~ tri afl~1b1ipOi~ f ~1t1 ~4middotmiddot f-gt -~ ~S-- ( -- - - _bull resin EactI billet then Undb~ tI1d prtiladlllltha5O __ eyilr
bull ~~_ ~~imiddot ~1iil h~ intlfcili~ ~eonbt1nlyi Tr1butylt1n F~id(18n)~~~O~dettu20i~blcicld8is 1IIfii bull i - ~ ( then 1IPI)l1ed to glvaadry Ulil tIi1c1ltnOss or 1(1)-150bullbull
tbull (2) IipreSSeCi Current Syat
bull A rigid ~tic due 1I8S drWedllliihwrtv-s41 liiIihOieS IIPProX~
bullbull 05-10 cia episrt ~ Pilnted ampUl8t Wsfbed IntaPQSIUon ampy
threadlng the nickel IIIlr8throu11 the dinto a tarproot junction
box Eliltilen of th8~1l1ii (recelving ~t-d curnritlere IIIlred
Into bull block connectOrrecelvlng dc trOll is potentiOstat The
rainJng iontroi blUets Wet8fbedfl11i1iiy lIIithln thejtlytlonbox
i~ityd~c IkiliPly ~~ ~lat1ruaWiode 8ncIacalOliel
refer~ electtodetoiere fllied ~ the per1aetat of the plut1cdlc IIIId
connected to the pot8ritlostat The desl~ of the potentlosbt ailoiolEld
~PlDteItlO1TClJrnnt to be MIllntained at 8prampset levtl of 966 II1II
c
r~
Grid raf 1J5s-t 114iH 563724 dud 0 Juna
Alolecule andbechr1a in general are ~ to
sutface lI1eroleyer (nlUSton) of _ter (~bullbull 191KJ)~To avold
contaJnatlonof ttw painted surf by U- -vents
blllets Were s-nact in a 1 conte1ner or atc1le (02i ) f~~nd
ter Iih1ch then f9ICI in tv 1IIhere the b~
be r-oved undeNeter El~~ bllleta fa Mcb trtai bullbullel
removed over a 1 dtq (336 hr~ expoeure perioda
each being ~~bn uposurnof 1 5 11 iA 168and DlIIauz1r
Arter field axpauHli the billets wan
fUtered _tar to ~ 1_ debtlflMd far 10 aJnuta ~
wv ~~~endattid sdbUiUd(otO
~ ~l f~ nn1ng el~~ --
lOOted onto 1middotCIi ~bullbullWIsend carbonmiddotcoa~nmiddotmiddot~middot f
each billet Ws~ Iis1ng a ccr1dge ~el~~
(5360) IIIlth anergy dJ8perslve X-ray alcroenelre1a (mx)
_lysis the les Wen reNed end coated III1th vold to al]Ql En
detailed sat of the alaquofa1l
(4) Lfiborattux ~
Painted taa~ bUletalllllre pnparad ~
in a Mrlne reclrculet1ng flOlll _ ~
f1Oll the ~ ~a1t~y Tha~~Un bullybullbullbull
10C M1nta1ned ~~t1as of ~t~la protazOllbull~bull10shy
(Entermap) ~ ~~~middotNlt~Eamp1~ __~and ~ r - ~ -
certaJn ~lc 1IQ1~ 1(1 the
the painted
~
thre repl1catft fn
plKed 1nto(022_)
- thIine1J~~ -lt bull
Atter mAX
(1-2) and i-n-t bn collected
14OC
Pain~
-
12 daY~ exposure and prepared as in d)~ - t
~I ~~~-- i~yen 1~ v
Rflltsi ~ _ ~f-~lI ft j bull _
rt~ll0~~~llY~~~~peln~stetsl~11i~ex~sed for
y~t1~J~ ~~~~1~1~~1UII~ya~kooJ~~~lt ~ ~~_~~I1J~ ~~~tomAX -lysis ~era than gold ~ bull bullbullbull c - _ bull lt
~~fllr ~~~ vllUll ~t1ans of the rfee htures and gt Co
~f~ - ~~~ ~f ~ ~aobta1Mdue IjIll811 iI1Flg 1~2 The IIIIdn ~~ ~ lt)ji9 )tmiddotmiddot - ht
- bull
~I rIIplt~ tIt8 -~ cOlllponanta of the pei1t 1bullbull WelIIlItr1x $ilj~t-~~middot - ~ _ -_
-IUch PVCbQed end rap~13y the chlorlna peak the
i~~~-~~-~V~~~-t18 ~~~~ ~~ Th8
ral~~i~ JlrQP(lrUo1sof ch 1Il-ntal paek~ll1d l~t_ Q~7 -gt- ~ _~ -- gt ) gt -_ -lt ~~ the 0 -end _ xprtsntabullTIwsa changes are -r1sed ~t~ lt~ _ ~ 1nT~ IThe8llia$lb npresentthe chengelI 1n~t1V8 p~lons
ti~~~ [ ~gt 3 - - - _ - _ -
Qf nellllllllt within We surface layars of the pe1ntmiddot ru The
~f~~thtJn -d ~rfeli avr the 336 h11cxI ampScanbe
SII4iIl r~ fig 3 al~ tht rta of~ dirfers tJetwaeo thaa ltmiddot~1- 1ft~- ~ _
IJOl appfiai-ed to ~1nlt1elly until ftar day 168 it fall
~~l~ 1h8relllllrelgn1fi~ diff~be~ qrOtactlid and
proieCted (at -966 ) piIIllnts tor lioo COPPer but not folimiddottin
Ibi~tlons Of the Pain -urface 8hoWad _Pil ~ witilt1M
(Plete I to IV)~ 1hePllat1na (~) pelnt(Phta x and b)
Clearl~st-i~~r alots on the SurfllC8 IiII1kh~ter1sed ~- f hlt1 lriitUl tin conCantrat1on~ lllariy slotsre Partially
~OIIe~ ~ tile _tr~tedal middot1lere rs ~an~ densely packed
grerltr~ ~th 1ar~(iIa1niy cotifIIr) 8ntt -ampniBil (lIB1rily 1rori)being
middot vl~l~~~~~ Dtt~i~~~t~ r~ ~f~~ I~t seawatllr for 11 n IIIOre slots appeared and th~ ~~ess _trb cwer
--if -- V~ c~~ f(i ~~~i~
(Plate Ie to It)~ the granules dld not protrude from the~rtace in ~ ~~~~j1~r~-ffJmiddot0~~
suhap~~r Both 0 nd --960 at ~~pa1nts ~ - fr ~~~~-w1-U~ ~~t
sl~llrat thi stag~ (Plates Ic-d II1d xrnspac~IlYh middotmiddotWith middot middotaJ~cmiddot gt incraallO ~alolre t~ difrrqcbullbull middotbatIiI8IImiddot the protected fjd
bull ~ ~ ~t~
IIllrotacbldpaint ~~lIfIt~ftitr~ h alQt~ p~t bull u bullbullbull J ~~i~~ 1~~
lIIith IIIuper lIdguon u 4lrotatad piIJnt (Pl8ti1 II) 1hD1II8S
Ill )1IMdw1lillS gnnJla~on f-r~~ dpert1cl ~ - - - ~~r ~t
Onthe 966 1IIpetnt t-var tIIO typeaofan with diffannt c t
~hIId~(Plate UI a to c) One hIId tIII-1 bull bullbull e _ bull
fUll wJ1hleas abvlaus alats the o1fwl bed C18UU1IlOt ~litUi middotC c ~ ~
less ~ t11 h Gnnulatlan __ but ~lIiIZly def1nlld in
middot~ latter Afte fila bed tiIIrtIId 10 ~are ~~j(
extansll(tlly ~ the~tIId(O at) tIIIIple lIIIdngthe~ - ~
middot orarallet1ai leas claaI (Plate I~) f u II1ao CQ1tJAJ11d to ~Gn - -
the -966 at 1 (Plata III d tor but 1JI8V8n 1nittNdcneabull -
Althatqa -middotslots wrs abvlaus becaI1ng 1 cwampr aiI- the orwWat1an8
f1nailyftar 14 day exposure ~ fUll drMIlapIIIent on all la - ~
are extensive with the 0 lhawlng both alota pore ~
OVIKgrOloln (Plata II c to f) Gnrulatlon 1bl to ~ gt~t~~ i
or the th1cknats of tho ma ~the ~ at ~ (~~~~ themiddot - - _ middott ~ ~ ~ - -1 1 t
middot f11bull IIIIIS IIOre extans1 thIIIn at dIlya but IIIIIIIY bull1Q~ tI st11l ~w -middotr~ 1gt ~1 ~
obvious fIIany ~ loI8re still p~ yen~~~~~~~r
than before Poiu appearedto be f~lf~ila~~c-~~~a (Plate IVf~ label 1) IIIIl1ch broke~i(2)~ ~~~~
t t
bull ~lS (3) ~ rflC~~ ~i~t~~l~~KI~ t~ldiIiiicyul~loIiih ~~Rht ~ )~- -~ - ~middotd 40 ( gt ~gt ~d lt1 t ~ Jt ~remiddot4amp tha paint trix cavity bottom eroded away reVe81~ng ore
t ~~t~ por~~ ~l~ P~]Y p~~8dby di~lutiaA~ ~~~~ti~Jr~gt~ta(l~~Siinll1cl1Ud~f iUiIiI reriatlGllbullt
bull a~~ ba~tid colCriAtloriccic~ GIl ~~~d ri -966 - ib~~~amp catQn1iti~~t~I~~(~t an ( ~
t the ~~~palnts (PIa_taiiI d~~h ~wh ~ n td have ~ CamplluJMitiIVc) ~~middottti o~lgtnalrf tlt of hIght
~ l=~A ~J~rW tYtles~ p~fiiilriQcOCCold to~Ja to jU-t ~jmiddoti~ai dlatClilS ~ir -jlati ~tbullbullent aNit ~ t ~~ th8middotf~ld~ies~~t~on~ ~ -966 prQ~~~(P14ta Iii~)Omiddotiliof~m1hiiHAiiipieat ere~ll-(piitir~ p-iot~iilans(PiA-iII~ end c) nbullbulltiledt dl~by1i~~ end wirinali1sheii ~cMI~bY thI tili
bull In t-~~ ~1i1nt 1oIIIeHfd~ ~m1IS (15OClind a t ri~r-dlatoa ~lor P~t dlataiOlonlt1anGIl ttw-966
t
bull ~
pr~t~ ~is~ih iion obitJauIJ A ~rot -=1 ~ ~ i __
reprasenttild on the 966 painted 1iUrfc1ncluicunljj ~ rwt
Achnanths Colonisation by bacteria 1n the laboretory bsed
bull eliperlMnt tollMct sbiUar pattern to the fild ~
bull bull Dilicueii
bull bull DUt 1Iidnt ~lona Mve dlttllrint ~~ ictian
It ~ JntM l1tenture ttt Iff yen __ JIll the
bull ~ bicins ani 111 ~t INchlnil (PhillIPs 1t73t EWene~ 1981
In ci1ntlHt tJWY ~tit ~Wl1c iIuCh as 1rbiltyl tirI
~ lite wAlII]ltiClr dlsP8tfu~ middotthiirsttilll~intaJn~ a
bull ~
lIIifoflll ampalCentraUon through the filII 10 thoIt biocide rel_ at tha
paint surfce 1fI$Jlts in IIIOre ditfusing ~s to repla~lj -iflJ~it
resulting 111 bullbulltNdy lr leechlng rata M1llat this IIIIY be middottruIi ot
tin ~ it was nOt true ~ ttw middotlBlf fOZWJlatiGn ua ~ bull - i - bull 1(1
ObSfvt1onundr SE clarly ~ tM t1l1 to be present
~ly lIS OIIIdle-llk parUc1~~t tn lUlfiice (Ito
IV) bUt usa ~ the dIpth attt fU-middot ~y~
If one considra the concn~rtlonmiddot chMge 111 1n ~111c COKICIl8rlts fro- the paint tU- i bullbullbull tMWei b~ ~ Si6t arid
tI-~t iron oxidt 1~ can bt (Fl_ 3 IrId TIibl8I) ~t ~ ase cona1derllble d1fftrenC8s ~ a Un decz bcdYMr ttw p1iod IIIIwraS lron~- an unt Ught inCr h ~ incqJper ~tlGn occUrnd _idly at ttw ~~ Gfb ~ period ~ out ttwn fu nPldly ampQII1n ~ umiddotirid II phast This IIIOUld bt tIlq)8Ctad It cantact l-=h1ng we occun1ng
IIltfIOS8d pertlcl of copper would lMch rapldly on t-n1on 1n - shy JIsewtr There would thlll1 ba bull J-a a1iltUy leu avalUba
i1 ~ ~
_tedal becfM uposed Thi tIOUld ttwn IIllCh Thla pulSing efttct
IIIOUld be expctad to -oth OIItlllith U- putic1 __ -lllible II tot 1~ GIl a on r-x _lao Thla teet 01 put1c1e 1tUng
can tit GIl the lIUrlace ClIlC8 it ~ a pan 1amp t
a 11aM bet in Plat I to IV IIaIrIg _ lit la_ II ~ 0- ~s _- PIate IW $llldl ~bull nshyot poRI flllNltttn Al~ tin ~ lou rete thtn I bullbullbull atill Ivwnce of bull al~t pulsed 1Itue rrlict tII1IIWd
elqlCtaG inIl ttwauUr ~ I I I
0
~ i
~ ~
~I J
bullbullt bullbullbullbullt bull bull ~
bull bull
I
bull t
~~
The same seqiHllle of cevity follliltloii (intttecaseofthetestjjltle
slot- shaped) fClliOileciby SecOlld8rYtlolesfotlllingin thll cavity
bottollis releasing further deep8rtox1ils (Plate XV d and g) As
cavities_re ProdICed ~ tiecaaIelnvadedby_ter~ onelilauld
expect 1ncreaa 1n1IOdha and dIladne lMls t the t1met
which the toxlM uw ~ Ths fCMld tCtHi eo (Flvmiddot 3)
~rttno the vi that_tat ~t1on otthe~ ~lY
il1Cl1l8S8S Idth UIiJ
Differences f~in toxin li~~to Ini 966 ill 8811P~ F ~ bOth Un end ~ lcMIr levels afillW
IIIIIlfI ptaInt(~1~~1ntha~~ ~~f~~ cst rklld with tbiIccpper ElJIc~l ta~Uona invalvinv
w pl8cId at a~ht~~middot3lO~lth Un cqJpel bull
(-960)10 ttW ~af this expei1tIIri1ts thIsiorelfJltdt -
that cqlp8r b8IiIId ~ IIiIOUldProc-ln f41ip1dly t thellllgllitiv
patentW _lied () as 1t 11 further f~1t ~ witt Un
III11ch is u-tat aqulllbd Idth
it If ttda 18 eo~ ~
~ )
frOll1 paintscould bl anqnced by an illprd current clithodic
protecUansysteipos1bly ahart_nino te 11fe of the paInts
antifouling ability Bacterial coloni Uon IIIU rtdUCed on the -s66 paint This
could have been due to unflSllOUlllble electrostatic cqrge on th surtece Th1a effect CIIl pley en IIIportant role in the t~of
bacteria to alrraces (lIIarsMll19E1O) As bacteria appear to IhiJw
greater toxicltyto copper (DMpsey 1981) it is Klre likely that
enhanced cqJper leacll1nl On the -966 AIIIPl c8used the recluCed
activity The IDlol8r toxicity ofUn to bacteria (as reported by
~ -
DelIIPsey Hle1gt or its lowes leaching rat was sUpPorted bythl
relatively rapid recolonisation of the slots by lIIicroorganiSlllS ~ftar thE inlt1al aurge of leachi~ Thailllpressecl current dto ~~ the oppoSite _ff~middot ~r on dtta colonstJon l1lI-1- in the
JIbcJrtory~ta larger rUDIIra f~ fit pro~td ~ther than uriprotected Pafnta
Ccnlus1one
1 tiild-a 18 presaIIUd ~ thattJn~ ~~
~llNChng notd1rf~ ~
2 TIleuseofCP~ ~CUIftnt(at) on1h1a ~fca41ng ~- ~
pa1I]t~ to ~_lAIIach1ngn_ at toxiN - -
3 leIJstaPidbectar1al ca1on1aatJon acaampmId an ~bId Pt1ntJ ~
4 JItore larger alcrofauJing (Oiat_ and PzOtoaoana) Gr~_
occurred on the protcbld~
I l
I
bull bull ~
mfEflpoundaS middot~-t9i _jgt- ~ _ _ ~sev M~J~ (1981) (cilonJzatiooof alUf~lng lnts by _rine
Jbull ~teda9g~1ta IIarina XXlV~ 185-191
- - gt 0 Ie 1 _ - _
poundV8I$i I~lf (1~P~h 1II8rin~Ugae and Foul lng AReviw ith
t p~l-i9Jrlr ~iiMdilto5hiP-fQUllfCiH 8ot~ ~r~ ~~~ XXIVbull lIPwl~J11A
Fotteth CN~R ~kk8nGB IWPIR1seJCiiiroaic1lihd ~e ~JIIth CII Offshore St1ucturu ElllS ~tjI~f~1Dt19i4
- - ~
UKC (1976) I~races ill ~W Ecology KIuverd Univbullbull middotraabullf~iddge ~bull C~~jII illt)Ia(t RC (1980) totinllsa Cattod1CPrOtect1cinbull middotmiddot8AtEmiddot~6haOCIlfif~ T~ p No 108 ~ 18 ~ 1IIHf bull
t bull lt ~J~~~ __ ~
Norkr4irulB(198D) Advanculn UcrOfj~lpound~ltiOyVol 4 (AlItxlaquoIder fl Ed) PlerIw Press lcxIdon~ lIP~ 51-85 ~ l~l - bull bull
PJijllt~i At(fm)~mr~~JnI~1fqul1nQ JlalntlJ Riumh ~JnOrpnic toatJngs 2 192middot
)~J gt0 c ~lt -- ~ __ RogerS~ Ut (1968) ina ~(lnt~fbItliP4ili GI COnoslon ~a Technology)bull bull
Sl~iorr~ VP~Ranald~ Rctd~~ (1S80)_ ~ Experllattal Stud1aiRelate middotEffect or Cathod1c Protict1Oi withmiddot terta1nr1c
Coating Syst~ 12th Annualofr T8Id1na1ogy Co1fItHnC8 liauston TIIlt p 9-12
j
fIG1 sPct FromMon IT8TJCuZOI Int Lliu EOAX
FlG~ Fro TlTCu20Ar-4 Dy Field Ellposult I~ir
il
I c
R hi -00amp=7
-I I I I I I I I I f
iii
bull bull
bullbullbull
I J
J
I Figur 3 Thl chan In reltl rqpoIQnlof -ariOtimiddotjOmponents of the paint film with time Thl vtic~1 ampkiS unl till _n ofmiddotmt Illment per unit OJ fe 50all Cl Na
Cu StI Feeshy
PlistinllFe S7~J6 704 3~74 908
f
r e ~ ee~
middotie ~lt - ~ shybullt bull ~
t -~ l Cu~ o ~ f
t --- 3igt - bull 0 J Sn
cc
t ~
~
bullo
~ bull e
bullbullbullbull bull ~
bull ebull a QI 1- bull 0 40 431282 316
101 dmllIII)bullt bullbullbullt
hr 0 ~~bulla 5~16 ~~ -~~ i166 4S i83 8~1B
24tu 6~480 SD~83
111middot 1~1 OS6
~ 5114 6~69 -51 UIamp middot016- lt ~rmiddot
1d o Mea 818 21 021
~Z21 199 1~1Qmiddot ~Jgt2 058
14 d o ~~ 804 Q~B8~~~a~~t~middot 685
866 372amp 8amp4 OS _i~11[ ~~64
Tttile 1 A81UvlL pzQJJortlcw 0 vxl~oI ~~bull Jn 1I1Dtitctel lt-1166) end lqIlDteCt6 (O)middot~af~_~middot_lodEDf~ middotmiddotmiddotmiddotmiddotmiddotmiddotmiddotmiddotc
~ ~
_----I~
J
I bull
I
J bull tmiddot
7o CONQRESOIMTERNACIONAL
I D~CORRO$IOr(IAAINAEmiddot~USTACIONES~v c)
t 7thflfERHATlONAL CONGRESS
ON IARINE CORROSION AND fO(JUNG
~CONGRE$INtERNATIONAL DE~CORR~ON ~E ET QES~SAUSSURE$
i t
~ ~
bullt
t t
UNIVERSIDAD PourtcNlCAVollncio7J1 NiwiemIif~ 1988
(SPAN
SECcl6lfn
BiOIogro marino Morine biologY ~1i1orine
Theunpub1ishedJllBIlu8criptEl have been reprod~d asr~ielved from the
authors The SCientiUc COlIIIBittee take not respoi~ulbll~~Y for any shy
error or omssi9n
Laa ponenciasno han side publicadas y estanreproducidas tal y como
sellan recibido demiddot BUSmiddot autorea pound1 Conte Cientfico middotno se responsemiddot
bil1za de cualquier error ~ om1si~n
bullbull _ bullbull_ bullbull ___ bull $_ _ L _ - _ --- -__
SESSION II Marine biology
Friday 11th November
SURFACE PROPERTIES ON NONmiddotTOXIC ANTIFOULING PAINT FILM
K Nanishi M Murase Y Yonehara M Kishihara and T Hirama
MICROFOULING ON ALUMINIUM PANELS PLACED IN THE TROPICAL WATERS
N B Bhosle K Venkat and A B Wagh
EFFECT OF TRIBUTYLTIN OXIDE (TBTO) ON THE OVARIAN MATURATION OF THE PRAWN CARIDINA WEBERI
R Nagabhushanam B Indira R Sarojini and Mary Frances Thompson
EFFECT OF BIOACTIVE MATERIALS FROM SOFT CORALS ON BACTERIA ASSOCIATED WITH COMMON FOULER
Vitalina Mary Avelin Mary R Sarojini and R Nagabhushanam
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
~ Deteiled experi~hts were iiIodeltwi~h DAP~he most effectiv~ corrosion inhibitor among thecoiaPoUndsstudiEiq The ett~cts ot
Ii its concentration antJ~mpe~ature in modifiction bath on the bull bullbullbullbull T _
Icorrosion rate of brass in an artificial see watet afe shown in Figs bull 1-3 Fig4 pres~ht8 the) effectir~ness of middotQlqMficationvs time of exposure for the saRIPls mOdified Wlth101-M DAP at 20middotC
- bullN tor 24 brs lt
I The gra1Dtetr1~ hur~~t~dtOJCuZlj20AJ2 middotmiddotallQY in the
stationary cQnditfonaevealed that tbecorroliion rate insee water was rdUcabi~bil~ttrtr~a~~ngtl1e brass ~ur1acbullbullmiddottdtil
DAP sQlut1onTlteettbullct otmiddotcconctfrClt-tonofthe DAFIIIOltl1fYing
I solutio~ upon th~ ~~~01l10it~~t~u ~oterY ~l~~At~O)DAP 11 protetedln~iMb~n usedirttbe concentntion 10~ M The
corrosiOnitf~~it~~~a~~~~~middot tji-~iticat10n tibullbee longer aM 10nger1beDAP$oludoMmiddot of concentrations O~5middot10lt
10or~0 tUled at~oejtb~Pdht ~ott~ct ~r increuedthe
J- corrOiti(gt~ ratoflt~OAI~ lllOY~ At t~etu-ra~dtolO~Cr tl14it oclif1cationwlth ui M
DAP swtaot1by i44U~edth~ corrOiJionzte Jbi ntwsmiddotre
lusindelIndent ~t the tioCII041f1ca1iCgtl themocl1iicatlonI ffectivbullnesswasin~bisc~C(~Jtib as ~a ebull Anlncreasein modification tempra~u~ middothllda~sitive effect oli corr~ionate also for D~SolUti~ri otconciitirationoSmiddot10-X bull
For DAP of concentrations 10 and 10 Il1O ledJdI bull nof sufficient iDlProvnt in resistance ~f the brass agAinSt
corroslon tas obee~e4even at elevated temperatures 060bull orr eomiddotc The rate of corrosion 4ecreasedyithincreasing JDOdification
I time and after the 24 hrs treatment it was nearly the slUDe as
that tor uIIIIOC1it iedbi-ass For the DPA S~lutionofconcentr~~ion10-middot the change in
bull modification temperature inthe range 60-80C did not produce
significant differeneilin corr~ionratesThe resultilUJ highly reduced rate of corrosion UCQGlpQred wi thufmodified slllllPles
J remained UnChangedindep~ndentl~ of modification time The lowest rote of corrosion was obtained for the slUDplea
I I
3
modified with DPAaolutionof concentration 0510-amp at 60middotC The protection effectiveness of 97 wae obtained after a 24 hr treatment Further increase of the solution temperature to aomiddotc gaveno ItIOre improveJpent
eonc1QSJons
nllt ~u1t presente~ ~ an be S~~S~l ~n the following conclusions
1 FrOID UIOng 11 compounds stucUed the Hst corros1on
protectioopro~ruH h~ 2-mercapt0-46-timinoPii1D1idiM (DAP) 2 The protection properties of DAF 1rovtd with increasing
rilodi t1cadonterature
3 DAP wu found to tie better corraaion inhibitor tor CuZri20A12 alloy than benzotrlazole
The lIOdit~cat1on effectiveness did not cbengewitllUme of exposition ot 8amP1es to an artificial see water
Ret-nc_
1 SpinellLP~middotMaj~hM BrunonoG Trabanel1LG r~t u
KDrro 31 919-923 (1980)
2CignaRG~oG BrCOZrobullbull J bullbull US 223-225 (1979) 3 WalkerR Corratoft 2Q 290-296 (1973)
4 Trabanell1G Zuchh1F bull BrunonoG Carru1t1V t Korrobullbullbull 24 602-606 (1973)
4
-- -II
JII
if ~-it CKpirons 101- dltf1lresect
it
it ~
coyio~rOIl tit Ctn20A i ii I uy if bH 5 dys ot
exposure to ltir tltl Cl d 1 see water ~ff~CdtioH condit 10IlsI)II1
it 10-middotmoled1W te_ 2o4Q6080middot
fi92 Therote of corrosion of CuZn20Al~ 01 aff~r 5 days ofit ex~EJuHl~ortif ictal 1-eet wateYKocfificiltrorP conditIons DAPit
10-middot ~ole~ temp 20406080middotC it Fi~~TIu(~~t ~f ~orro~ionmiddot~f cuiP20A2~111i) otter 5 days of it ~~~~I~iii~~rtIti~i~l s water Moaiflcafiorl ~ohifitians DAP
~ 4 ( 1- -~~ ) ~~ II10-~ IIioledIII temp 20406080 C I ~
Fi~4 The lIlQCl~t ica~iol) effec~iven~s Yt exposure time torIt CuZn20Al2 alloy in ortiflci~t slaquolelfaterbull KoclificaqQnc~pditions
bullI it
~ Dllt~i~lQmiddot~Jel~ telllP 2PmiddotC~l hr~
ibull -
bullbull bullt
bullJ
bull bullt
bullbull 5
bullt
~ i ~
12bull 5 p
3 6
r1g 1
t bull 2OC iIrC to-c tIOC
-24
16 iMtiMtillle Ihl
I I
~ N
t ~ 5bull
JOC bull AOc bull IiOC -we I
I I I
3 1amp l I modifiCation tiMe Ihl
I I
t
f ~
_--
yshy-~~
-1shy~~(~
Jl -
- _JIJ 11 _ bull ~ bullbull L IIIJIIM
I bullbullbull shy
-
if Ilt
JII l3$i]J ~e-l~ ~ 1
J ill KItt
Ii I 1 8 Ij j iJ)Sj
emiddot Sllll0 r B 8Je Do Ji
t ~l u Ji ]
1 0
j
j
r 1l3iiiJail1h
bull III
~ i J ii i )1 t~I ~
~
~ )38 ttl i UN ~I3 11 I ~iii bullbull I fi~fi IelI M 6 6 bull s plj i I Jjhi
~l -u II11 i~ Ji ~aJ j~ ~ tll
- bull
-------------- -- - --- -- - -- shy
2
This paper desr1blS the use of ti1c~lctrooes to measure pHbull bull _ bullbull ~- l bull bull
iJQ2 chmgtS ~~ 1II1C~olones ~middotilIlg~ ~f~~s i~rsed ll ~ter ~ wlJlch t1)U11ncjotgaI11sllls are grolog SOllie p~tnt1al 11P1~~tions C1f~se cIIatlges to corrosloO at the
lt I )
surlP alllO br1lJfl)dl~ - 1 - ( _ ~ -
ItIiterWs 8nd ~ ~ ~
() poundI~~lt - -
Ttl lec~_ 1n theu bpelsnts hIM bnndeScdbed In
~~~I )iel~~bull~~i~) but n 11 ~ haJe
- Aj ~-
(1) pHel~
ThapHalec~ lII8rII basadQIJ tne-iWof It1tQ (1__)
wre the pHlt1V8recenton of~~l~ 1a fiI)OS8d bull 1Ihart po1nt extlndlngfro- tt lnsulatlnQ (Flljj1) The
pH andmiddot alUlllno~nlcate glau 1cropipettesued lnthe - - ~
electrode constnction went ~ using bull wrt1cal alectrode
puller assntielly bull vart~c81 version of the equiplllltnt
orlQlnally developed by Al8Xllllder and NasWlaquo1953) The ~ ~ -
electrolyte u-d1llllS 1N tel and filllnQ IIIIIS 8aoIpllshad using bull ~ - or
aydnge 8nd a vary fine draIIIn out Qlass capillary
Once f1lled the elfCttocJes were soaked for severalday~ in 1111
tel to allow coqlletehYdroi~s ofthe pH glass lin lnternd refelflf1(e of sllver lrecoated wi til sliver chlorld~~~s
ccinmctedmiddot to a gold plated pin iealed into the imd of the
electrode lII1th~ill~oo rubber COMPleted ei~trodei er~ stated Up downIIIards lna dilute solution of ~r~iIcrcj In allmiddot uper~ middot-temal reference Qf bull~~l
allCtradilllllS UIIIId DurlnQ -~ts -n bulllact~damiddot was c~cted to ind~ ~~F~Y(_ itrgtbull The cStcu1t layautforbotitthiiptl lIlIdpDz electtaifes
~
1a IIhIWi Sn Figure 2 bull
~ -~ - ~ -- - - - I -
CaltbnUariof all alacbodn id Clirr1ed middotOutmiddotbllfGn ftc AIIICPUIIIri inmiddot c- elctmdeiimiddotmiddotmiddot_middot~
ckalng en apars-nt by in nJIalt~ orthey dldriat
pEOCia ~lbblIt ~lntal callbraUon In ~
CIIIIi8S all ~ datafl(WtMI middotmiddotdl
(11) 0ItygIn a1acUocfes bull
TIUI catheter typa elachode use a fiat ended and of duw 1 to 15 _ It conslatad ofa ~ uia P~tJft- wlra
aeelad 1n 9l~ and IIIDadded in 1IPaKY~1o 1I1th a aUler wue KUng a raferance (see FiQ3) Ihen the epOxy hId~
h I of h l~middot careMJy ~ flat to UpQM
bath the plau _the ~lwr 1IIIIIbnre WIis ttn8llPlled
ecrosa ~ flat lind by rep8atedly d1fIPlng Jt into a aQ1utJDn Of
collodion 10 atheta1cohol A Pola1-1slng voltage of m all __
applied bit eachelectrodl and these re not thariuSed for
axper1lllental purposes until middotthe response I118sstllblallihen
bullp
4
bullit tuffer elut1on a~ a eorustllllt
it i)
~~ ~ ~eted IItld accepted for uSe elliCtlodes
it lt gt ~ ~~J ~J _ bull~ - - -gt bull
IIIIiIr~~Jn~IIOWt1on of 02 KCl D~ygen Ilectrodeser _iliry~ ~~)~ 5middot -1 ~ J~- - f -
it cli~rated aga1pst solution I of known oxygen content (as 1 w f - w ~-l _ bull bull
i IllllllaUred bY n of the l111nkler titration thad APHA 1915) ~Lmiddotmiddotlmiddot ~ ~
it Any el-=trodI not lIlCtIib1ting a ilnaar response rejected ~ ~ ~ ( _ - ~
zelia axyg8n lution obteined by the usa of a buffer
i aturated wUhpyrogallol crubbed nitrogen The 100$
~~~ reed1ng obtained by bubbling a1r thlOUltl the ~ ~~bull ~ ~ bullIII burfal far 12 hours at ~~ture 8M pressure- ~t bull
(b) [)cpars-ttal~
All -rs-tts IIIIIl1I Wried aut in atellperature controlled
bulli it
tank pliad an a teflain NYler ted l~ steel patfon ill insidil a ane _tre CIbI Faraday CIlIIIi(fig 4) Tbe~
prav1decI IIf1th a lJrfQII hinged door at the fiant for eccns and[f as gro~nded to earth to scrbullbullnout radio frequencyi intelfuwce The _tar flowing ttmluitI the experluntali - tri pullPed at a controlled rate fro a temperature
~ controlled reservoir For all experillanta involving algae
i llllA1nat1on provided using daylight fluorescent twes if l1d teel and 1Ial1ns plywood plate approxilaately 20 bull x 15
wire Ixposed in the -rine environmantfar several IIIQIIths to bull
allow a heal) growth of IIIICrofoul1ng organi_ to deVelop The
bullI steel plates had been placed utldally and IIIItre exclu8ively
bullt
~~==1$ and ~klOOrganl_ ~ldirig Itltena ~srar-nt of their
OWl The wooden plates 1IIIIIl8~sn the Jntett1ifalteclJQii
-f llllllre COIIered lIIIith ta 01 tt- alp InWfCld1IIPtth
occa1a1onal bryozaens There IIIIIIl8 IWIo lagII quantitibullbull of
~ retained -gat the algal fu-nt nPiate lIIIIIle alsO cleansed and stalJil1aad_ bllton ~
provided On 12haUrly~ ~dllrUght luar~w1 RddtUOlllll steel plat8s ~~In Alii ~dIxk~ lit
Uwrpaol (lie) IIIhare they IIIIIIl8 co~ by middot~Of~
(xtllus edulis) and the lISCld1M Clona intUn1l us
~1IIIIIl8 roved f the pJaqa NP~1id In cl_
ta tor 5 days at cansqnt ~ toIitDUampe
pH WId pOz ~1ane frc the extuMl t~~
the fouling to the Milstrata llllllre ncarded tt elecUadM
middotheld in a rigid pttllpex ~rr1et attechld to II ~ly
Jointed GIl on the lI1crcIIanipulAtor To nciad the muM the
tnraugh the foulJng tI1IIIIerds the ~ta JI_ta 0( 1 _
could be acCUllitaly controlled by talng the tIM clrlw of
the n1pJlator Iftn each lllc~ the DVIII ~oIdlft9 shy
allowed to _tabIU for ane wte prior to ncordJng the
VGltge~
~
bull a
-lt 1
l~to the_ batnacl~ 1ls~ COC)trol - fmiddotmiddotmiddot
recod4l9sof W pH 8I)d pOi at the SIJIIEdepth outside tOe 5hell middot-middotvmiddotmiddotmiddot_I~ lgt~middot middotmiddot~_middotmiddoti 1gt )
wete made
-flesolts ~
(a) pH IIIldIDz p~lllllS
The _re~ts for Hancl p02proflle tluough both algal -d - ~ gt Comiddotmiddot i -
eniIIIalfou11ng are gi~ in Figs Sto8bull --
TjpicalProtUeis daIiilttuougha t of Entronlorpha are ~1n -bull l J ltl ~~ -
Fig 5 - lIhllsttyp1cal d1innal fluctuations CICCUIJ8d during bull ~i bull bull e bull bull
photosynthetiC- II1d non-phOtinwnthet1c lods (Fig Sa) mad _~ d q _ _ __ _ i bull
decreases inpHlIIIIl1I~ IIIIa) fro-theaul-f1lCe of ttWt bullbull bull ~ bull
dcyenI tqwards the abstratacn tdch it enclOSlild (fig 511)
~fal1s1n pH--nted to as-ach as 2 units Over 9 IIIj
dIpth
figure Sa ahoid the thangea in pH -d Pl2 between 11ve barnacles
with depthln a stlat1c envirornent A steady decrease in ~
occurred as the plata IlUrfe~es toiereappro8ched pHWtially
fell on one profile before rislngbut lOSe steadily with daPth
on the second praflle SlIdlar changes are IIt10wI in fig 6b
with depth into the shells of dead barnacles 80th oxygen -d
pH levels were re~at1l1E1ly steadY ~ similar to the 8IIIblent - ~- lt
envlrOlWOt Until the -actual dead shell IIias entered When a
IIIUked drop in both fJ02 and pH CICQ11Tedbull
~llell in rigs rit JllellllUrellt$IIere maltle ~
iOlfillidual IlUs$els withtiRle (Fig 7dlb) In thll first
experisent elactrodes were placed indicated (fig)
arCUld indivicUllalsln genU flawJog ~ter The
resUlts show bull ~tle fIwrease ~ IID2 eMIS bull aiJc hour 1Ucj
The pH~t of the~ II4ttIn ~~atmiddot~
81 (te UlbientpH being 80) but 1GII8 ateIIfIU ~ the
1rtIa1ent aJphon to a6bull 111 iIadIIr ~~thI aff~ of
flOiW 00 local ~ciIndltJans~ the ~
~tadbut tile flow at~ turned Off tar 25i1aur
Jadin thelllddleofthlt ~The nailts g1R
in FlQIb -PD2 IIMtls mae ~y inflQldng CCnd1tJanamiddotbut
fell -zkadly fter alhoit lao w flaws caadawJnit a
~ fall lit 90 alii Levels rapidly ratond onca t IIIIUtchadDli agaJil No ILICh teed varlauon ~
in pH around the eldlalent 1pbn 1iIIflImI a taIdyJncn4IM -
obsmt8d aspec1elly cleM to thIIen1Ml CondltLni around
indivOJa1 ~ lIIhllst of interest are lMa repl_tative
of field conditions fro around groups or clIIIPS The
IlICpIIr1ants were ~ ~ted Wittdn denSe clUIIPs of ala
fouling a plate These r8SIJlts are given 10 figs tie (1Il1Ud
IlUlIS8ls) -d 8b (within byssal thrtiIds) In tatic ccinditJans
amprCUld the _15 u-a1(8$ ~ fellrapldly (45l) reelJ pft
rose sl1rjltly and then feU aftar 2 h llhen flowing condltiors
were resuraed after J hthe ~ rose to the original middotlevels shy
The pH rose lArlcedly at the eltictrodenear to the xhalent
bull bull bull bull bull
bull11
bullbullbullbull
it
I
8
s~ (3) but reriialned stlaadyahd ultilllateiy fell silghtiy (2)
inttie genetal fooling inassbull
~
III1thihthe1JyMli~~ Or~selci~5 IiIfieretilltti($trltus teniis ltO -=IiUlteliter~ ~ ~(Flg 8b) ~ lIIithrrolaw~ cCincentreuoos diCIppea framp1~D lBigtif3S
~bUt toaitnpl~lylt4i 150 IIifIo lIIhfjnficw t8st0ted~middot pH
t wi~1rithi thread ClUIps~lMl1BtO 74 (3) IinIt B~ito
785(2) lri-staUc cinfit1at IoihUst ettn88dgtiofthlt ~
it -flnedf81r1y~at aroOOd17S~ lIIlanf1Dlilwu
rellitoteilpft inltleUyJose (by 04 UI1ts at Ii1ta3) but it theo
bull fall only to startrfs1riq tgi1nattM not theIlXper1ilent~middot
bull Dlseusslont f tMresulblmiddot obta1nIid it ~be cl8lirly sean that f(IIJUngI
bull species vethe abUlty to Gtly IIOdlfy thdr loclmiddotmiddot
bull InIl~ts
bullbull L1t1t end dark period tluctuat1ona in algalts IBre obsarved
end ware lri close wpeeIIIen~ with those reported by Terry end
bull ~
Edyv- (1981) with 1ncnasaln both pH end IID2 llIrJngllttt
traquor1ods lIhilst thlshappened ttM surface of the t pre
bull fU_ts ware frelyprojecting into UJe water -deep intottie
mat nearer to the solid _stratlli whl1lre shading fll1lli light andbull accuIIJlatlonmiddot of deconIposJng ~gallIIBtllrial DCCIINed pH valuesbull fell IIIBrkedly Concurrent wlth this dlQp in pH ilaS II fell inbull
bullbull
9
oxygen Such t~ral and $PiItial va11ations in ~andpft
would certainly qive 11se to corrosion cells middotatmiddot the sUbsttatie
surface ~ly resulting in iorroaien pitting
The r] _tabellc ctivlUee or fau)Jng -=raJrwerttmlbullbull
together with their prapeN1ty to-=ci-1tritla~
tlwlsalves wIll also lIied to 1dcIoenv~ lIOdifktIona
Resplretory activities wU1 tend to ~~ tn-the tft and Input caapanants auch a cinia~nY qatk
1mIertabJatesmiddot ara mlk 1bullbullbull middot the bulk of their nlUagllnoua
wastes ~ excrated as ~ 1heexpftunt ncord~ pDz
end pH cningS amN 1nd1v1Q8l~ ~ ~ri
claady 1nd1cat41 the ~ of _tft~t GIl tt ~ of lnIirollliental changes Inmiddot expr_~ta lllhere no Wlter
--nts occurradthmi a np1d dacreUfi1ll pDz
COIJIdalo be a ~decreae in pH (no a) v fladrlg
condlt1C1ns are r-ed tIwn Ieymiddotmiddotor iIay nat be a 1bIIeIqI1int PH
1nctease dBpwIding ~ the --- ofmiddot iheltr ~ by the
organl1n the cluIIp 0wIgn wui a influenced by the
proxJa1ty of 1nhalent or ~a~ the dlnsity IIId the
tIlJcknes$ of the clullp w111alsa t)IMt wiuence in ~
wetr -N_tbull Hencein thick clampllP aUt and detritus
cCUlllulatlon is are COIIIIiIn ~ theh billa IIIdfbls
restricted Br8IJIIltdow1 gf organtc datrital _ter1als CI3IIIIUIIS
oxygen resulting in recLlcing conii1Uons llleasurlllenta haw
shown that sulphids qulckly build up In such deposits and
sulphate recLlcing bacteria y tie-~ SRBconcentrtlonmiddotin
J -I
II II IJI Ii ill
II
-bull I
11
10
the depOSlts woUld terid to Ile lOW at We start of ~r1ments as
condit1onswere nat favourable (POstgate 1979) The cantlooQUs
prockIctiPn of r1th f~plieUdofeacal_te1alor IlUlpllonill1l
~ts bYpoundilt8~ (~~Iick1ngand 1IkIod1) hOweva ~ t H 1 (
IIIOUld rep1dly altAIr theli11l~t and the rua~ofSRB re
notedtofncreasemiddotttcllll ( 1 SRaIll1taat llaUt 1 x 101 SMIIl-1
aftet bull -th
IUctotJlIIl corrosion canfe=t _ta1sln a1Ullberof Ways
(Iverson1972 fUlllIt end King 1975f 1Uilr 1981) pH can
aff~ the ectivity of the 1IIin organlasinuolved the H 1IIho
hava tin qlu pH range beban 10 12 (Graff 1981)
although tIdS -can be lnf1u8nCed by ather fllltors In atI1tl11n
they lIKIJlre a catbon end nitrogen aurCa 11 various
salts tin alactron accaptor (HedJart ~ idbullbull 1985)bullAil of
U- facton are availabll in the debris -alltting b41tween
ar1na foullng -d the 1lUbstra~ 110 providing bull potantlally
corrosiveanvirorwent
Evan IIIhete such accullIUlatiori does not take place varlable
changas inoxYQ8ll and pH ~trations acrossthe suface of
teel soch as C3I becaustd by foul~ ~ C3I give ti_
to corms1an calla lalildlng to piftlng corrosion
There is an initial tllld8llCY for the pH to rise as a relUlt of
the excretoryactivi1i~ of fouling aniIIal and this 1amp UIIllkely
to give rise to corros1an problems Eventually ~er feacal
h
1
~I
materials and detrltus~ill build up around or~anisms ~re bo~
low pH and p02 devul9ls providing a potentially coJJOampiIu
envinx-nt flaw does dify thh _build UfJ but inmiddotthbull
Ixparlunts carrllldotttid notprlvl(lt it R~~r it
l8Iilated the depth of deposit ~1~~ in t- ~ It ahoutd alao r~~- in f~ population so-1QIJvlWela 111111 be dying or dUd Thlb
~lng~wUl~1Ielacal1Md conoa1ve ~iccandltiorJs-t SR8a 111111 b4I fCllllfC314lng ~y pOIi1desa hlbitt~ HtInd 1tt~ WIi~ul create 1~1Mct Jrcx-ntal dlffarenca ~ ~th l1kel1haOd or pitting
n
References
Albilnder J~T 1 bL~1953~~(9EthII prockIctiPn of IdcroaltJodIIs UIIId in I~ studies Rev Sc1 lnstra ~ 528-531
_r1can Ptbl1c Health laSoc1ation 1975 5taIIdIrd~for the lIIIIIlystS of _tars end taatms 11lth pounddIt1on 1IPHIi-~ 1015 ~taenthSt NW 1IIa-l~~OC 2IJ]J6 USA
Bau-8laquo=k1ng C~IIIG amp 1IIoOd EJF1955 Biologlcal ~ in theestuarlna envi~t 1 EcolClil) of the ~ cycle Kcnlridlkjl Ned Akad -t ~ Ser B~ fi 160-181
cter DB amp SUver IA 1961 In IRafaranctlElectrodl$l EdltolS Iva DlG and l-z Gl lc8dilll1c Pr London Maw Yodlt Chapter 11 ~
Davies PIII I 8r1ric middotF 1942JII1c1oel4iitrodas for-aln local oxygen tension in n11 tissues Rlv Sci InStruII Yo 524-533
EdyileariRJGJ ~ Tetty LA UJ83 The 1nfluenc ~lIlcro algae on corrosion ofstructural steels used 4l~ the 1tIirth
Sea 81odeterioratian sect 336-347
bull bull
~
bull - -- ~It Jf bull
- ~ 13
12
Terry LA I~Ed)VeangtRfiJ 1981 lIi(toal~IOI1CGrmianF~ l)i 1St1oJhemailnEifooiiiigffixed gftshoreIt Bot 1Iar ~ 117-183 lsect~~jlaUoQ1l1 _ ~t Ef)e~gy Offs~r1I Te~lOtjjy Paj)8r
-middotltcntpI)~it$t~l91-16PP~middotmiddotdl ( I
bullbull
iIIalk e1 JL 1971 bull Ion specfie Uquid I~~ e)(charig~ ndcroelactroclels Anal ChIIaI ~8S-Q2 AGraf1~ w~S 9S1~ 1orrosloh InFIntroooctI00 tooffshOle
51~weSoes~~fab~lcat~onIns~allat1on Gulf Wall PI aelUnger EG 1982 The appl1~tlon 9fMl1ikIngmiddotHoust(jfW~Q23g-258middot icroelectrade in the study of pH end PD2 in lIHinli
HerbierC SN fGH~rt~ )RI -Stoctcdalpii Watltif1Sal RS ~1ronIIInb Ub~~ 27l-Z79 1985 Fac~or lontro1l11)9 tha activity ofmiddot sulpMte
bull Wall P 1983 The appllcatlon Md ~ ofiwrJ)e~c~odl shy-taicfii9 tiiCtetili tn~ diiilncj tar infection
techniques for the iiaurnt cif icraeov1ror1liltital5QiCiety ofJebo1- Engi~ SPE 139781-10 _i~~ - ~ ~ i ( _ IKIdif1catlons caued by aGII8 rlnefou~lng -C~ebullbull
PhD lhaaia Ibiverslty or ~tar UKbull bullbull bull -
IIH1nke JA 19fiSa In EXperllRants in Physiology and middotampioClIiIiiilatJift l c E(If(orltGAiKetkutgt ~c-Presa-
Wooltdngton AObullbull Davenport J 1983 pH lind IlIl2 IMla ffl~- j~t~5 -lMIIleath 1IH1na ro-fauUngGqWl J bull Expbull 1
B101 Earlsectsect 113-124 Klnke JAIII 1969b II) Glass IUcroelectJQdes poundx1tors Uwal1~Ui~ OrOllt arid~t Nt JOhn fII11ey YOI1lt thaptar 11
~-f _f~ ~ ~~nt ~ Igt
bull Iverson IIIP 1972 Biolog~ Qrroaian In Advencesin
corrosion scianca and tecIYIology 2EdS FCntaia G~ I bull IIS~et RIIIbull p~ PJaSsLancIoIl 1-42
bull
Jorgensen BB Reusbech NP I Cohen T 19B3 Photosynthesis and strUcture of bIIn~ II1crobWp MC1OIIlectrocki and ~ stud1ea of four cyanabacledalt -~usllWtOl ~bullbull 1~f093
t l~ O~ ~~~ H~ ~9s7~c-gtHatste~iecfn1k van shy pellediftlrtenPt - Stlchelektraden (1-Su
AiJBitiKlificfiliilislflilr)zlIr polrapusdiailJIIissui1g_ liIserstoff(jruQkas fur d1eB lIsUbullbullung der IIliJlttOUf1cillatlon PflftgaHArdI ~fl39
bullbull Mller JOA 1981 tals In EconcinI1c II1croblolooy sect McNbUdbicldatariOtationEd Rosebull A middotAcadelc Praulondcn149-202
t Mller J DAI King RA 1975 8io$~iorat1on of lllltals In lIIi~1al ClflPlCts of the deterloration of tarWbullbull EdsGilbertRJ~ middotloveloCk 0111 bullAcadeic- Presst IILandon 8Jo103
bullt
IPotgate JR 1976 The sulphate-reducingbacteda
Callbridge tIn1versity Press Cliilbrldg8U-K
Reusbecb fUI i Ward DIII 1964 i~~~1Ida5tud1el of
bullt inter$Utilll IIIItez dttm1stry and photosynthetk activity
in ilflaI spring Idcrob1altAppl~EIi~ MerobW tsect4IlU-27lt I
IIbullt bullt
bullbull Ibullt
1
Inmiddot middotmiddot11middotmiddot f~~ a~ -tmiddot bull J~f
I~ W
i5iImiddot J~a t
ilmiddotmiddotmiddot I middotIt Ii
l
~
middotmiddot~~middotlL= g r
Lj Ic
t
i ~
I
gi II cEi ~ I I
1 t rW
D
tR bull
bull t 10
11
r
i
I ~
j
~
I
~bullbullbullbullbullbullbullbullbullbullbull
LV
FigS pH and p02 profiles into macrofoltling eomlllllllitilS dominated by Balm trenatU$ on steel plate 11 two pH profiles Ib) two lIOai profiles leI pH and P02 profiles into dead barllllCle shiella withdetri~Us I I
itIbl r p02 ~ I
nvnHg 11 70
j ifJi eo 1amp0
laquof it 30 s
I 20 N
ca 11 10 a o -- shy i bull
11
80
19
781 o 2 6 8 10 12 0 2 6 amiddot 10 12 dIpdI Qf ~iftto fouling In HighIst numbIII pIatIi
8deg~~
~i c5 iioo5 fiil
pH 8 bull_ ~
~~ ~
_ bullbullbull- pH
-pOz
(cJ
e-Clf ( j~JJlaquo1- --~I ~a ilil - _ I i ]1
~ ri en i ~ oJt ~iJ id tUI
2u1U I shyI t
shy1 2 3 6 6 7 8 9 10 11 12 i shyDepth 9f electrode (mm) into fouling
1 i
1
~----------~-------
~
~-~ lt1~
middotmiddotPmiddot ~
I
V
~
ill
1 Tbellitt~~ of cathodiC protection OO~~lIic~tlO aridrfOflllUlc
ofi AA~Uoullr14l ~1~I
I
ael~lner EG middotIUtcb8l1AJ Dept lt4 lInY1roMe1lal Bl01011
=~~venilt7 IlariCheW 1f139PL $ ~11al1df
r
I
Ablitrict ~~~ e~l billttI ~ted with ~pert1n ant11_1111ampp paint llere itpoii~1n _vatI- In the tl~ld iUIit 1bOioatbrt~ScM Were
1IUbjll~ to qtbocJ~C ~teltt1on byen _ current (~)SJi inci EDAX Iieamp~ta IJdlctecI ~ lachln teoc copper OIl tbePlCtec~ pa1n~ atter14 dQ1a~ protected Pllnt abo ebowed
) reduCed bacteiial activIty butmiddot ~e4 dltoe Iu1d protOzoanmiddot co~~l~tlont~1l IlU l~byenCOCltact not byen dUfwIl00 ~r _baRl lrillolvedln tOXln nil ereio noted
l ~~
1
I
Introduction
Applications of protective coatings and cathodic protect1on(cp~l
are recognised corrosion control methods for IIIBrine steel Itructureso Cathod~c protectlOll preventl conCIsion by Int~lng til8ctrl~l
currentS trem externalllClUfCa to e~~tthlJno~ e~ carrollon feetlans C04U lUChu tUauJ1ng Aelnts a
(
barrIer to ~ the flao of couos1a1 ~aboIHnu~r
Dccurrlng anodes cathodes or wlwit gelvWc CCIIII1 ( 19681
flinger 1980) AnUfauling ~ also JJICOqIOrII~ b1oc~~
Tr~ylt1n nuor~ (~) end o~ Oxide (~) (~ flJ1J
- of u blacld1111~
V _Ut Evans 1981) ~ ~
end growth of bipfoullng 0I1I)IIIII1IIIIe Wdd~ ca ~ of
paint rusendmiddotmanemiddot ~(r~ til )
As corrosian ~t1on ~ bICCIIIII -an ~1tlQd ~ of
antiroullnQs 1 conjunction _ith CP have bllClIOr ~
resultIng in reseudt 1rwestllj111tlng trw COIIIPIitlbUlty or VU~ ~t t _~
rOratlation to dlfrert levI ot CP (~Eelf 18110 ~
~t 1980) RelatIvely 1nIestiljlllUans have ~ urtw ~
or CP an thIt anUraullngpert~ of palnta
In thIt preseot 1nlIestioaUon tV~iln111 ronutlon ud
cOIIIHrclallY incanjlntlon wIth CP was ~t4IC~ Iw I sraed current (-966 l1li) arid exposed at the 1 Strait IInQlbullbullbully
~t1ons weremiddot 00 the IIhcIrt bnifects the PprIUd_
had an a) the various COIIIPQI1eIIts of n pe1ntf1l11 end b) l1oft~
developaentbull
bull bull ~
~ ~ - f
bull ~ _T~~-middot~ ~~- ~r _ -$1
(1) ~rlttionor Painted Steel Billets middot_ ~~tmiddot_tlJ middot~hmiddot~--~fmiddot ~ middot~A~-~~middot - -~ ~
Thirty-al ateel blllets (8-10aD2 end l depth) were cut frollbullt ~middot~middot~middotmiddot~middottrSmiddott ~jS-~ It bull gt~ tfl - middot--imiddotmiddot ~ t1dItrd steal plates The dimenSlonsof -thB butits-lim chosento
bull -~~ middotmiddotgt2h ~ ~-Imiddot ~~rt ~- -middoti~~ ni ~~middot1 l middot-~t~~middot _
(a) fecilltate amiddot Ie detrucUve -thxI of prePraUonpdor to ~~_gt gt~ k7i ~_ t~ 1--1- _ ) - +--_~ -_~bull ming electron ~ 4II1CI (iialloWdirectobs8iVatlCnS of the
- i lt-- ~ gtbull -o1e billet 8Uface
gt r~middotmiddotj ~L(~middot~middot rmiddot tmiddot J ~ - ~ - lt _ bull Single non-standard (1hIula~) nickel lIIireSmiddot Spot Welded e1Cing
bull ~1deJ ~ b~i~ ~j~~ tri afl~1b1ipOi~ f ~1t1 ~4middotmiddot f-gt -~ ~S-- ( -- - - _bull resin EactI billet then Undb~ tI1d prtiladlllltha5O __ eyilr
bull ~~_ ~~imiddot ~1iil h~ intlfcili~ ~eonbt1nlyi Tr1butylt1n F~id(18n)~~~O~dettu20i~blcicld8is 1IIfii bull i - ~ ( then 1IPI)l1ed to glvaadry Ulil tIi1c1ltnOss or 1(1)-150bullbull
tbull (2) IipreSSeCi Current Syat
bull A rigid ~tic due 1I8S drWedllliihwrtv-s41 liiIihOieS IIPProX~
bullbull 05-10 cia episrt ~ Pilnted ampUl8t Wsfbed IntaPQSIUon ampy
threadlng the nickel IIIlr8throu11 the dinto a tarproot junction
box Eliltilen of th8~1l1ii (recelving ~t-d curnritlere IIIlred
Into bull block connectOrrecelvlng dc trOll is potentiOstat The
rainJng iontroi blUets Wet8fbedfl11i1iiy lIIithln thejtlytlonbox
i~ityd~c IkiliPly ~~ ~lat1ruaWiode 8ncIacalOliel
refer~ electtodetoiere fllied ~ the per1aetat of the plut1cdlc IIIId
connected to the pot8ritlostat The desl~ of the potentlosbt ailoiolEld
~PlDteItlO1TClJrnnt to be MIllntained at 8prampset levtl of 966 II1II
c
r~
Grid raf 1J5s-t 114iH 563724 dud 0 Juna
Alolecule andbechr1a in general are ~ to
sutface lI1eroleyer (nlUSton) of _ter (~bullbull 191KJ)~To avold
contaJnatlonof ttw painted surf by U- -vents
blllets Were s-nact in a 1 conte1ner or atc1le (02i ) f~~nd
ter Iih1ch then f9ICI in tv 1IIhere the b~
be r-oved undeNeter El~~ bllleta fa Mcb trtai bullbullel
removed over a 1 dtq (336 hr~ expoeure perioda
each being ~~bn uposurnof 1 5 11 iA 168and DlIIauz1r
Arter field axpauHli the billets wan
fUtered _tar to ~ 1_ debtlflMd far 10 aJnuta ~
wv ~~~endattid sdbUiUd(otO
~ ~l f~ nn1ng el~~ --
lOOted onto 1middotCIi ~bullbullWIsend carbonmiddotcoa~nmiddotmiddot~middot f
each billet Ws~ Iis1ng a ccr1dge ~el~~
(5360) IIIlth anergy dJ8perslve X-ray alcroenelre1a (mx)
_lysis the les Wen reNed end coated III1th vold to al]Ql En
detailed sat of the alaquofa1l
(4) Lfiborattux ~
Painted taa~ bUletalllllre pnparad ~
in a Mrlne reclrculet1ng flOlll _ ~
f1Oll the ~ ~a1t~y Tha~~Un bullybullbullbull
10C M1nta1ned ~~t1as of ~t~la protazOllbull~bull10shy
(Entermap) ~ ~~~middotNlt~Eamp1~ __~and ~ r - ~ -
certaJn ~lc 1IQ1~ 1(1 the
the painted
~
thre repl1catft fn
plKed 1nto(022_)
- thIine1J~~ -lt bull
Atter mAX
(1-2) and i-n-t bn collected
14OC
Pain~
-
12 daY~ exposure and prepared as in d)~ - t
~I ~~~-- i~yen 1~ v
Rflltsi ~ _ ~f-~lI ft j bull _
rt~ll0~~~llY~~~~peln~stetsl~11i~ex~sed for
y~t1~J~ ~~~~1~1~~1UII~ya~kooJ~~~lt ~ ~~_~~I1J~ ~~~tomAX -lysis ~era than gold ~ bull bullbullbull c - _ bull lt
~~fllr ~~~ vllUll ~t1ans of the rfee htures and gt Co
~f~ - ~~~ ~f ~ ~aobta1Mdue IjIll811 iI1Flg 1~2 The IIIIdn ~~ ~ lt)ji9 )tmiddotmiddot - ht
- bull
~I rIIplt~ tIt8 -~ cOlllponanta of the pei1t 1bullbull WelIIlItr1x $ilj~t-~~middot - ~ _ -_
-IUch PVCbQed end rap~13y the chlorlna peak the
i~~~-~~-~V~~~-t18 ~~~~ ~~ Th8
ral~~i~ JlrQP(lrUo1sof ch 1Il-ntal paek~ll1d l~t_ Q~7 -gt- ~ _~ -- gt ) gt -_ -lt ~~ the 0 -end _ xprtsntabullTIwsa changes are -r1sed ~t~ lt~ _ ~ 1nT~ IThe8llia$lb npresentthe chengelI 1n~t1V8 p~lons
ti~~~ [ ~gt 3 - - - _ - _ -
Qf nellllllllt within We surface layars of the pe1ntmiddot ru The
~f~~thtJn -d ~rfeli avr the 336 h11cxI ampScanbe
SII4iIl r~ fig 3 al~ tht rta of~ dirfers tJetwaeo thaa ltmiddot~1- 1ft~- ~ _
IJOl appfiai-ed to ~1nlt1elly until ftar day 168 it fall
~~l~ 1h8relllllrelgn1fi~ diff~be~ qrOtactlid and
proieCted (at -966 ) piIIllnts tor lioo COPPer but not folimiddottin
Ibi~tlons Of the Pain -urface 8hoWad _Pil ~ witilt1M
(Plete I to IV)~ 1hePllat1na (~) pelnt(Phta x and b)
Clearl~st-i~~r alots on the SurfllC8 IiII1kh~ter1sed ~- f hlt1 lriitUl tin conCantrat1on~ lllariy slotsre Partially
~OIIe~ ~ tile _tr~tedal middot1lere rs ~an~ densely packed
grerltr~ ~th 1ar~(iIa1niy cotifIIr) 8ntt -ampniBil (lIB1rily 1rori)being
middot vl~l~~~~~ Dtt~i~~~t~ r~ ~f~~ I~t seawatllr for 11 n IIIOre slots appeared and th~ ~~ess _trb cwer
--if -- V~ c~~ f(i ~~~i~
(Plate Ie to It)~ the granules dld not protrude from the~rtace in ~ ~~~~j1~r~-ffJmiddot0~~
suhap~~r Both 0 nd --960 at ~~pa1nts ~ - fr ~~~~-w1-U~ ~~t
sl~llrat thi stag~ (Plates Ic-d II1d xrnspac~IlYh middotmiddotWith middot middotaJ~cmiddot gt incraallO ~alolre t~ difrrqcbullbull middotbatIiI8IImiddot the protected fjd
bull ~ ~ ~t~
IIllrotacbldpaint ~~lIfIt~ftitr~ h alQt~ p~t bull u bullbullbull J ~~i~~ 1~~
lIIith IIIuper lIdguon u 4lrotatad piIJnt (Pl8ti1 II) 1hD1II8S
Ill )1IMdw1lillS gnnJla~on f-r~~ dpert1cl ~ - - - ~~r ~t
Onthe 966 1IIpetnt t-var tIIO typeaofan with diffannt c t
~hIId~(Plate UI a to c) One hIId tIII-1 bull bullbull e _ bull
fUll wJ1hleas abvlaus alats the o1fwl bed C18UU1IlOt ~litUi middotC c ~ ~
less ~ t11 h Gnnulatlan __ but ~lIiIZly def1nlld in
middot~ latter Afte fila bed tiIIrtIId 10 ~are ~~j(
extansll(tlly ~ the~tIId(O at) tIIIIple lIIIdngthe~ - ~
middot orarallet1ai leas claaI (Plate I~) f u II1ao CQ1tJAJ11d to ~Gn - -
the -966 at 1 (Plata III d tor but 1JI8V8n 1nittNdcneabull -
Althatqa -middotslots wrs abvlaus becaI1ng 1 cwampr aiI- the orwWat1an8
f1nailyftar 14 day exposure ~ fUll drMIlapIIIent on all la - ~
are extensive with the 0 lhawlng both alota pore ~
OVIKgrOloln (Plata II c to f) Gnrulatlon 1bl to ~ gt~t~~ i
or the th1cknats of tho ma ~the ~ at ~ (~~~~ themiddot - - _ middott ~ ~ ~ - -1 1 t
middot f11bull IIIIIS IIOre extans1 thIIIn at dIlya but IIIIIIIY bull1Q~ tI st11l ~w -middotr~ 1gt ~1 ~
obvious fIIany ~ loI8re still p~ yen~~~~~~~r
than before Poiu appearedto be f~lf~ila~~c-~~~a (Plate IVf~ label 1) IIIIl1ch broke~i(2)~ ~~~~
t t
bull ~lS (3) ~ rflC~~ ~i~t~~l~~KI~ t~ldiIiiicyul~loIiih ~~Rht ~ )~- -~ - ~middotd 40 ( gt ~gt ~d lt1 t ~ Jt ~remiddot4amp tha paint trix cavity bottom eroded away reVe81~ng ore
t ~~t~ por~~ ~l~ P~]Y p~~8dby di~lutiaA~ ~~~~ti~Jr~gt~ta(l~~Siinll1cl1Ud~f iUiIiI reriatlGllbullt
bull a~~ ba~tid colCriAtloriccic~ GIl ~~~d ri -966 - ib~~~amp catQn1iti~~t~I~~(~t an ( ~
t the ~~~palnts (PIa_taiiI d~~h ~wh ~ n td have ~ CamplluJMitiIVc) ~~middottti o~lgtnalrf tlt of hIght
~ l=~A ~J~rW tYtles~ p~fiiilriQcOCCold to~Ja to jU-t ~jmiddoti~ai dlatClilS ~ir -jlati ~tbullbullent aNit ~ t ~~ th8middotf~ld~ies~~t~on~ ~ -966 prQ~~~(P14ta Iii~)Omiddotiliof~m1hiiHAiiipieat ere~ll-(piitir~ p-iot~iilans(PiA-iII~ end c) nbullbulltiledt dl~by1i~~ end wirinali1sheii ~cMI~bY thI tili
bull In t-~~ ~1i1nt 1oIIIeHfd~ ~m1IS (15OClind a t ri~r-dlatoa ~lor P~t dlataiOlonlt1anGIl ttw-966
t
bull ~
pr~t~ ~is~ih iion obitJauIJ A ~rot -=1 ~ ~ i __
reprasenttild on the 966 painted 1iUrfc1ncluicunljj ~ rwt
Achnanths Colonisation by bacteria 1n the laboretory bsed
bull eliperlMnt tollMct sbiUar pattern to the fild ~
bull bull Dilicueii
bull bull DUt 1Iidnt ~lona Mve dlttllrint ~~ ictian
It ~ JntM l1tenture ttt Iff yen __ JIll the
bull ~ bicins ani 111 ~t INchlnil (PhillIPs 1t73t EWene~ 1981
In ci1ntlHt tJWY ~tit ~Wl1c iIuCh as 1rbiltyl tirI
~ lite wAlII]ltiClr dlsP8tfu~ middotthiirsttilll~intaJn~ a
bull ~
lIIifoflll ampalCentraUon through the filII 10 thoIt biocide rel_ at tha
paint surfce 1fI$Jlts in IIIOre ditfusing ~s to repla~lj -iflJ~it
resulting 111 bullbulltNdy lr leechlng rata M1llat this IIIIY be middottruIi ot
tin ~ it was nOt true ~ ttw middotlBlf fOZWJlatiGn ua ~ bull - i - bull 1(1
ObSfvt1onundr SE clarly ~ tM t1l1 to be present
~ly lIS OIIIdle-llk parUc1~~t tn lUlfiice (Ito
IV) bUt usa ~ the dIpth attt fU-middot ~y~
If one considra the concn~rtlonmiddot chMge 111 1n ~111c COKICIl8rlts fro- the paint tU- i bullbullbull tMWei b~ ~ Si6t arid
tI-~t iron oxidt 1~ can bt (Fl_ 3 IrId TIibl8I) ~t ~ ase cona1derllble d1fftrenC8s ~ a Un decz bcdYMr ttw p1iod IIIIwraS lron~- an unt Ught inCr h ~ incqJper ~tlGn occUrnd _idly at ttw ~~ Gfb ~ period ~ out ttwn fu nPldly ampQII1n ~ umiddotirid II phast This IIIOUld bt tIlq)8Ctad It cantact l-=h1ng we occun1ng
IIltfIOS8d pertlcl of copper would lMch rapldly on t-n1on 1n - shy JIsewtr There would thlll1 ba bull J-a a1iltUy leu avalUba
i1 ~ ~
_tedal becfM uposed Thi tIOUld ttwn IIllCh Thla pulSing efttct
IIIOUld be expctad to -oth OIItlllith U- putic1 __ -lllible II tot 1~ GIl a on r-x _lao Thla teet 01 put1c1e 1tUng
can tit GIl the lIUrlace ClIlC8 it ~ a pan 1amp t
a 11aM bet in Plat I to IV IIaIrIg _ lit la_ II ~ 0- ~s _- PIate IW $llldl ~bull nshyot poRI flllNltttn Al~ tin ~ lou rete thtn I bullbullbull atill Ivwnce of bull al~t pulsed 1Itue rrlict tII1IIWd
elqlCtaG inIl ttwauUr ~ I I I
0
~ i
~ ~
~I J
bullbullt bullbullbullbullt bull bull ~
bull bull
I
bull t
~~
The same seqiHllle of cevity follliltloii (intttecaseofthetestjjltle
slot- shaped) fClliOileciby SecOlld8rYtlolesfotlllingin thll cavity
bottollis releasing further deep8rtox1ils (Plate XV d and g) As
cavities_re ProdICed ~ tiecaaIelnvadedby_ter~ onelilauld
expect 1ncreaa 1n1IOdha and dIladne lMls t the t1met
which the toxlM uw ~ Ths fCMld tCtHi eo (Flvmiddot 3)
~rttno the vi that_tat ~t1on otthe~ ~lY
il1Cl1l8S8S Idth UIiJ
Differences f~in toxin li~~to Ini 966 ill 8811P~ F ~ bOth Un end ~ lcMIr levels afillW
IIIIIlfI ptaInt(~1~~1ntha~~ ~~f~~ cst rklld with tbiIccpper ElJIc~l ta~Uona invalvinv
w pl8cId at a~ht~~middot3lO~lth Un cqJpel bull
(-960)10 ttW ~af this expei1tIIri1ts thIsiorelfJltdt -
that cqlp8r b8IiIId ~ IIiIOUldProc-ln f41ip1dly t thellllgllitiv
patentW _lied () as 1t 11 further f~1t ~ witt Un
III11ch is u-tat aqulllbd Idth
it If ttda 18 eo~ ~
~ )
frOll1 paintscould bl anqnced by an illprd current clithodic
protecUansysteipos1bly ahart_nino te 11fe of the paInts
antifouling ability Bacterial coloni Uon IIIU rtdUCed on the -s66 paint This
could have been due to unflSllOUlllble electrostatic cqrge on th surtece Th1a effect CIIl pley en IIIportant role in the t~of
bacteria to alrraces (lIIarsMll19E1O) As bacteria appear to IhiJw
greater toxicltyto copper (DMpsey 1981) it is Klre likely that
enhanced cqJper leacll1nl On the -966 AIIIPl c8used the recluCed
activity The IDlol8r toxicity ofUn to bacteria (as reported by
~ -
DelIIPsey Hle1gt or its lowes leaching rat was sUpPorted bythl
relatively rapid recolonisation of the slots by lIIicroorganiSlllS ~ftar thE inlt1al aurge of leachi~ Thailllpressecl current dto ~~ the oppoSite _ff~middot ~r on dtta colonstJon l1lI-1- in the
JIbcJrtory~ta larger rUDIIra f~ fit pro~td ~ther than uriprotected Pafnta
Ccnlus1one
1 tiild-a 18 presaIIUd ~ thattJn~ ~~
~llNChng notd1rf~ ~
2 TIleuseofCP~ ~CUIftnt(at) on1h1a ~fca41ng ~- ~
pa1I]t~ to ~_lAIIach1ngn_ at toxiN - -
3 leIJstaPidbectar1al ca1on1aatJon acaampmId an ~bId Pt1ntJ ~
4 JItore larger alcrofauJing (Oiat_ and PzOtoaoana) Gr~_
occurred on the protcbld~
I l
I
bull bull ~
mfEflpoundaS middot~-t9i _jgt- ~ _ _ ~sev M~J~ (1981) (cilonJzatiooof alUf~lng lnts by _rine
Jbull ~teda9g~1ta IIarina XXlV~ 185-191
- - gt 0 Ie 1 _ - _
poundV8I$i I~lf (1~P~h 1II8rin~Ugae and Foul lng AReviw ith
t p~l-i9Jrlr ~iiMdilto5hiP-fQUllfCiH 8ot~ ~r~ ~~~ XXIVbull lIPwl~J11A
Fotteth CN~R ~kk8nGB IWPIR1seJCiiiroaic1lihd ~e ~JIIth CII Offshore St1ucturu ElllS ~tjI~f~1Dt19i4
- - ~
UKC (1976) I~races ill ~W Ecology KIuverd Univbullbull middotraabullf~iddge ~bull C~~jII illt)Ia(t RC (1980) totinllsa Cattod1CPrOtect1cinbull middotmiddot8AtEmiddot~6haOCIlfif~ T~ p No 108 ~ 18 ~ 1IIHf bull
t bull lt ~J~~~ __ ~
Norkr4irulB(198D) Advanculn UcrOfj~lpound~ltiOyVol 4 (AlItxlaquoIder fl Ed) PlerIw Press lcxIdon~ lIP~ 51-85 ~ l~l - bull bull
PJijllt~i At(fm)~mr~~JnI~1fqul1nQ JlalntlJ Riumh ~JnOrpnic toatJngs 2 192middot
)~J gt0 c ~lt -- ~ __ RogerS~ Ut (1968) ina ~(lnt~fbItliP4ili GI COnoslon ~a Technology)bull bull
Sl~iorr~ VP~Ranald~ Rctd~~ (1S80)_ ~ Experllattal Stud1aiRelate middotEffect or Cathod1c Protict1Oi withmiddot terta1nr1c
Coating Syst~ 12th Annualofr T8Id1na1ogy Co1fItHnC8 liauston TIIlt p 9-12
j
fIG1 sPct FromMon IT8TJCuZOI Int Lliu EOAX
FlG~ Fro TlTCu20Ar-4 Dy Field Ellposult I~ir
il
I c
R hi -00amp=7
-I I I I I I I I I f
iii
bull bull
bullbullbull
I J
J
I Figur 3 Thl chan In reltl rqpoIQnlof -ariOtimiddotjOmponents of the paint film with time Thl vtic~1 ampkiS unl till _n ofmiddotmt Illment per unit OJ fe 50all Cl Na
Cu StI Feeshy
PlistinllFe S7~J6 704 3~74 908
f
r e ~ ee~
middotie ~lt - ~ shybullt bull ~
t -~ l Cu~ o ~ f
t --- 3igt - bull 0 J Sn
cc
t ~
~
bullo
~ bull e
bullbullbullbull bull ~
bull ebull a QI 1- bull 0 40 431282 316
101 dmllIII)bullt bullbullbullt
hr 0 ~~bulla 5~16 ~~ -~~ i166 4S i83 8~1B
24tu 6~480 SD~83
111middot 1~1 OS6
~ 5114 6~69 -51 UIamp middot016- lt ~rmiddot
1d o Mea 818 21 021
~Z21 199 1~1Qmiddot ~Jgt2 058
14 d o ~~ 804 Q~B8~~~a~~t~middot 685
866 372amp 8amp4 OS _i~11[ ~~64
Tttile 1 A81UvlL pzQJJortlcw 0 vxl~oI ~~bull Jn 1I1Dtitctel lt-1166) end lqIlDteCt6 (O)middot~af~_~middot_lodEDf~ middotmiddotmiddotmiddotmiddotmiddotmiddotmiddotmiddotc
~ ~
_----I~
J
I bull
I
J bull tmiddot
7o CONQRESOIMTERNACIONAL
I D~CORRO$IOr(IAAINAEmiddot~USTACIONES~v c)
t 7thflfERHATlONAL CONGRESS
ON IARINE CORROSION AND fO(JUNG
~CONGRE$INtERNATIONAL DE~CORR~ON ~E ET QES~SAUSSURE$
i t
~ ~
bullt
t t
UNIVERSIDAD PourtcNlCAVollncio7J1 NiwiemIif~ 1988
(SPAN
SECcl6lfn
BiOIogro marino Morine biologY ~1i1orine
Theunpub1ishedJllBIlu8criptEl have been reprod~d asr~ielved from the
authors The SCientiUc COlIIIBittee take not respoi~ulbll~~Y for any shy
error or omssi9n
Laa ponenciasno han side publicadas y estanreproducidas tal y como
sellan recibido demiddot BUSmiddot autorea pound1 Conte Cientfico middotno se responsemiddot
bil1za de cualquier error ~ om1si~n
bullbull _ bullbull_ bullbull ___ bull $_ _ L _ - _ --- -__
SESSION II Marine biology
Friday 11th November
SURFACE PROPERTIES ON NONmiddotTOXIC ANTIFOULING PAINT FILM
K Nanishi M Murase Y Yonehara M Kishihara and T Hirama
MICROFOULING ON ALUMINIUM PANELS PLACED IN THE TROPICAL WATERS
N B Bhosle K Venkat and A B Wagh
EFFECT OF TRIBUTYLTIN OXIDE (TBTO) ON THE OVARIAN MATURATION OF THE PRAWN CARIDINA WEBERI
R Nagabhushanam B Indira R Sarojini and Mary Frances Thompson
EFFECT OF BIOACTIVE MATERIALS FROM SOFT CORALS ON BACTERIA ASSOCIATED WITH COMMON FOULER
Vitalina Mary Avelin Mary R Sarojini and R Nagabhushanam
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
-- -II
JII
if ~-it CKpirons 101- dltf1lresect
it
it ~
coyio~rOIl tit Ctn20A i ii I uy if bH 5 dys ot
exposure to ltir tltl Cl d 1 see water ~ff~CdtioH condit 10IlsI)II1
it 10-middotmoled1W te_ 2o4Q6080middot
fi92 Therote of corrosion of CuZn20Al~ 01 aff~r 5 days ofit ex~EJuHl~ortif ictal 1-eet wateYKocfificiltrorP conditIons DAPit
10-middot ~ole~ temp 20406080middotC it Fi~~TIu(~~t ~f ~orro~ionmiddot~f cuiP20A2~111i) otter 5 days of it ~~~~I~iii~~rtIti~i~l s water Moaiflcafiorl ~ohifitians DAP
~ 4 ( 1- -~~ ) ~~ II10-~ IIioledIII temp 20406080 C I ~
Fi~4 The lIlQCl~t ica~iol) effec~iven~s Yt exposure time torIt CuZn20Al2 alloy in ortiflci~t slaquolelfaterbull KoclificaqQnc~pditions
bullI it
~ Dllt~i~lQmiddot~Jel~ telllP 2PmiddotC~l hr~
ibull -
bullbull bullt
bullJ
bull bullt
bullbull 5
bullt
~ i ~
12bull 5 p
3 6
r1g 1
t bull 2OC iIrC to-c tIOC
-24
16 iMtiMtillle Ihl
I I
~ N
t ~ 5bull
JOC bull AOc bull IiOC -we I
I I I
3 1amp l I modifiCation tiMe Ihl
I I
t
f ~
_--
yshy-~~
-1shy~~(~
Jl -
- _JIJ 11 _ bull ~ bullbull L IIIJIIM
I bullbullbull shy
-
if Ilt
JII l3$i]J ~e-l~ ~ 1
J ill KItt
Ii I 1 8 Ij j iJ)Sj
emiddot Sllll0 r B 8Je Do Ji
t ~l u Ji ]
1 0
j
j
r 1l3iiiJail1h
bull III
~ i J ii i )1 t~I ~
~
~ )38 ttl i UN ~I3 11 I ~iii bullbull I fi~fi IelI M 6 6 bull s plj i I Jjhi
~l -u II11 i~ Ji ~aJ j~ ~ tll
- bull
-------------- -- - --- -- - -- shy
2
This paper desr1blS the use of ti1c~lctrooes to measure pHbull bull _ bullbull ~- l bull bull
iJQ2 chmgtS ~~ 1II1C~olones ~middotilIlg~ ~f~~s i~rsed ll ~ter ~ wlJlch t1)U11ncjotgaI11sllls are grolog SOllie p~tnt1al 11P1~~tions C1f~se cIIatlges to corrosloO at the
lt I )
surlP alllO br1lJfl)dl~ - 1 - ( _ ~ -
ItIiterWs 8nd ~ ~ ~
() poundI~~lt - -
Ttl lec~_ 1n theu bpelsnts hIM bnndeScdbed In
~~~I )iel~~bull~~i~) but n 11 ~ haJe
- Aj ~-
(1) pHel~
ThapHalec~ lII8rII basadQIJ tne-iWof It1tQ (1__)
wre the pHlt1V8recenton of~~l~ 1a fiI)OS8d bull 1Ihart po1nt extlndlngfro- tt lnsulatlnQ (Flljj1) The
pH andmiddot alUlllno~nlcate glau 1cropipettesued lnthe - - ~
electrode constnction went ~ using bull wrt1cal alectrode
puller assntielly bull vart~c81 version of the equiplllltnt
orlQlnally developed by Al8Xllllder and NasWlaquo1953) The ~ ~ -
electrolyte u-d1llllS 1N tel and filllnQ IIIIIS 8aoIpllshad using bull ~ - or
aydnge 8nd a vary fine draIIIn out Qlass capillary
Once f1lled the elfCttocJes were soaked for severalday~ in 1111
tel to allow coqlletehYdroi~s ofthe pH glass lin lnternd refelflf1(e of sllver lrecoated wi til sliver chlorld~~~s
ccinmctedmiddot to a gold plated pin iealed into the imd of the
electrode lII1th~ill~oo rubber COMPleted ei~trodei er~ stated Up downIIIards lna dilute solution of ~r~iIcrcj In allmiddot uper~ middot-temal reference Qf bull~~l
allCtradilllllS UIIIId DurlnQ -~ts -n bulllact~damiddot was c~cted to ind~ ~~F~Y(_ itrgtbull The cStcu1t layautforbotitthiiptl lIlIdpDz electtaifes
~
1a IIhIWi Sn Figure 2 bull
~ -~ - ~ -- - - - I -
CaltbnUariof all alacbodn id Clirr1ed middotOutmiddotbllfGn ftc AIIICPUIIIri inmiddot c- elctmdeiimiddotmiddotmiddot_middot~
ckalng en apars-nt by in nJIalt~ orthey dldriat
pEOCia ~lbblIt ~lntal callbraUon In ~
CIIIIi8S all ~ datafl(WtMI middotmiddotdl
(11) 0ItygIn a1acUocfes bull
TIUI catheter typa elachode use a fiat ended and of duw 1 to 15 _ It conslatad ofa ~ uia P~tJft- wlra
aeelad 1n 9l~ and IIIDadded in 1IPaKY~1o 1I1th a aUler wue KUng a raferance (see FiQ3) Ihen the epOxy hId~
h I of h l~middot careMJy ~ flat to UpQM
bath the plau _the ~lwr 1IIIIIbnre WIis ttn8llPlled
ecrosa ~ flat lind by rep8atedly d1fIPlng Jt into a aQ1utJDn Of
collodion 10 atheta1cohol A Pola1-1slng voltage of m all __
applied bit eachelectrodl and these re not thariuSed for
axper1lllental purposes until middotthe response I118sstllblallihen
bullp
4
bullit tuffer elut1on a~ a eorustllllt
it i)
~~ ~ ~eted IItld accepted for uSe elliCtlodes
it lt gt ~ ~~J ~J _ bull~ - - -gt bull
IIIIiIr~~Jn~IIOWt1on of 02 KCl D~ygen Ilectrodeser _iliry~ ~~)~ 5middot -1 ~ J~- - f -
it cli~rated aga1pst solution I of known oxygen content (as 1 w f - w ~-l _ bull bull
i IllllllaUred bY n of the l111nkler titration thad APHA 1915) ~Lmiddotmiddotlmiddot ~ ~
it Any el-=trodI not lIlCtIib1ting a ilnaar response rejected ~ ~ ~ ( _ - ~
zelia axyg8n lution obteined by the usa of a buffer
i aturated wUhpyrogallol crubbed nitrogen The 100$
~~~ reed1ng obtained by bubbling a1r thlOUltl the ~ ~~bull ~ ~ bullIII burfal far 12 hours at ~~ture 8M pressure- ~t bull
(b) [)cpars-ttal~
All -rs-tts IIIIIl1I Wried aut in atellperature controlled
bulli it
tank pliad an a teflain NYler ted l~ steel patfon ill insidil a ane _tre CIbI Faraday CIlIIIi(fig 4) Tbe~
prav1decI IIf1th a lJrfQII hinged door at the fiant for eccns and[f as gro~nded to earth to scrbullbullnout radio frequencyi intelfuwce The _tar flowing ttmluitI the experluntali - tri pullPed at a controlled rate fro a temperature
~ controlled reservoir For all experillanta involving algae
i llllA1nat1on provided using daylight fluorescent twes if l1d teel and 1Ial1ns plywood plate approxilaately 20 bull x 15
wire Ixposed in the -rine environmantfar several IIIQIIths to bull
allow a heal) growth of IIIICrofoul1ng organi_ to deVelop The
bullI steel plates had been placed utldally and IIIItre exclu8ively
bullt
~~==1$ and ~klOOrganl_ ~ldirig Itltena ~srar-nt of their
OWl The wooden plates 1IIIIIl8~sn the Jntett1ifalteclJQii
-f llllllre COIIered lIIIith ta 01 tt- alp InWfCld1IIPtth
occa1a1onal bryozaens There IIIIIIl8 IWIo lagII quantitibullbull of
~ retained -gat the algal fu-nt nPiate lIIIIIle alsO cleansed and stalJil1aad_ bllton ~
provided On 12haUrly~ ~dllrUght luar~w1 RddtUOlllll steel plat8s ~~In Alii ~dIxk~ lit
Uwrpaol (lie) IIIhare they IIIIIIl8 co~ by middot~Of~
(xtllus edulis) and the lISCld1M Clona intUn1l us
~1IIIIIl8 roved f the pJaqa NP~1id In cl_
ta tor 5 days at cansqnt ~ toIitDUampe
pH WId pOz ~1ane frc the extuMl t~~
the fouling to the Milstrata llllllre ncarded tt elecUadM
middotheld in a rigid pttllpex ~rr1et attechld to II ~ly
Jointed GIl on the lI1crcIIanipulAtor To nciad the muM the
tnraugh the foulJng tI1IIIIerds the ~ta JI_ta 0( 1 _
could be acCUllitaly controlled by talng the tIM clrlw of
the n1pJlator Iftn each lllc~ the DVIII ~oIdlft9 shy
allowed to _tabIU for ane wte prior to ncordJng the
VGltge~
~
bull a
-lt 1
l~to the_ batnacl~ 1ls~ COC)trol - fmiddotmiddotmiddot
recod4l9sof W pH 8I)d pOi at the SIJIIEdepth outside tOe 5hell middot-middotvmiddotmiddotmiddot_I~ lgt~middot middotmiddot~_middotmiddoti 1gt )
wete made
-flesolts ~
(a) pH IIIldIDz p~lllllS
The _re~ts for Hancl p02proflle tluough both algal -d - ~ gt Comiddotmiddot i -
eniIIIalfou11ng are gi~ in Figs Sto8bull --
TjpicalProtUeis daIiilttuougha t of Entronlorpha are ~1n -bull l J ltl ~~ -
Fig 5 - lIhllsttyp1cal d1innal fluctuations CICCUIJ8d during bull ~i bull bull e bull bull
photosynthetiC- II1d non-phOtinwnthet1c lods (Fig Sa) mad _~ d q _ _ __ _ i bull
decreases inpHlIIIIl1I~ IIIIa) fro-theaul-f1lCe of ttWt bullbull bull ~ bull
dcyenI tqwards the abstratacn tdch it enclOSlild (fig 511)
~fal1s1n pH--nted to as-ach as 2 units Over 9 IIIj
dIpth
figure Sa ahoid the thangea in pH -d Pl2 between 11ve barnacles
with depthln a stlat1c envirornent A steady decrease in ~
occurred as the plata IlUrfe~es toiereappro8ched pHWtially
fell on one profile before rislngbut lOSe steadily with daPth
on the second praflle SlIdlar changes are IIt10wI in fig 6b
with depth into the shells of dead barnacles 80th oxygen -d
pH levels were re~at1l1E1ly steadY ~ similar to the 8IIIblent - ~- lt
envlrOlWOt Until the -actual dead shell IIias entered When a
IIIUked drop in both fJ02 and pH CICQ11Tedbull
~llell in rigs rit JllellllUrellt$IIere maltle ~
iOlfillidual IlUs$els withtiRle (Fig 7dlb) In thll first
experisent elactrodes were placed indicated (fig)
arCUld indivicUllalsln genU flawJog ~ter The
resUlts show bull ~tle fIwrease ~ IID2 eMIS bull aiJc hour 1Ucj
The pH~t of the~ II4ttIn ~~atmiddot~
81 (te UlbientpH being 80) but 1GII8 ateIIfIU ~ the
1rtIa1ent aJphon to a6bull 111 iIadIIr ~~thI aff~ of
flOiW 00 local ~ciIndltJans~ the ~
~tadbut tile flow at~ turned Off tar 25i1aur
Jadin thelllddleofthlt ~The nailts g1R
in FlQIb -PD2 IIMtls mae ~y inflQldng CCnd1tJanamiddotbut
fell -zkadly fter alhoit lao w flaws caadawJnit a
~ fall lit 90 alii Levels rapidly ratond onca t IIIIUtchadDli agaJil No ILICh teed varlauon ~
in pH around the eldlalent 1pbn 1iIIflImI a taIdyJncn4IM -
obsmt8d aspec1elly cleM to thIIen1Ml CondltLni around
indivOJa1 ~ lIIhllst of interest are lMa repl_tative
of field conditions fro around groups or clIIIPS The
IlICpIIr1ants were ~ ~ted Wittdn denSe clUIIPs of ala
fouling a plate These r8SIJlts are given 10 figs tie (1Il1Ud
IlUlIS8ls) -d 8b (within byssal thrtiIds) In tatic ccinditJans
amprCUld the _15 u-a1(8$ ~ fellrapldly (45l) reelJ pft
rose sl1rjltly and then feU aftar 2 h llhen flowing condltiors
were resuraed after J hthe ~ rose to the original middotlevels shy
The pH rose lArlcedly at the eltictrodenear to the xhalent
bull bull bull bull bull
bull11
bullbullbullbull
it
I
8
s~ (3) but reriialned stlaadyahd ultilllateiy fell silghtiy (2)
inttie genetal fooling inassbull
~
III1thihthe1JyMli~~ Or~selci~5 IiIfieretilltti($trltus teniis ltO -=IiUlteliter~ ~ ~(Flg 8b) ~ lIIithrrolaw~ cCincentreuoos diCIppea framp1~D lBigtif3S
~bUt toaitnpl~lylt4i 150 IIifIo lIIhfjnficw t8st0ted~middot pH
t wi~1rithi thread ClUIps~lMl1BtO 74 (3) IinIt B~ito
785(2) lri-staUc cinfit1at IoihUst ettn88dgtiofthlt ~
it -flnedf81r1y~at aroOOd17S~ lIIlanf1Dlilwu
rellitoteilpft inltleUyJose (by 04 UI1ts at Ii1ta3) but it theo
bull fall only to startrfs1riq tgi1nattM not theIlXper1ilent~middot
bull Dlseusslont f tMresulblmiddot obta1nIid it ~be cl8lirly sean that f(IIJUngI
bull species vethe abUlty to Gtly IIOdlfy thdr loclmiddotmiddot
bull InIl~ts
bullbull L1t1t end dark period tluctuat1ona in algalts IBre obsarved
end ware lri close wpeeIIIen~ with those reported by Terry end
bull ~
Edyv- (1981) with 1ncnasaln both pH end IID2 llIrJngllttt
traquor1ods lIhilst thlshappened ttM surface of the t pre
bull fU_ts ware frelyprojecting into UJe water -deep intottie
mat nearer to the solid _stratlli whl1lre shading fll1lli light andbull accuIIJlatlonmiddot of deconIposJng ~gallIIBtllrial DCCIINed pH valuesbull fell IIIBrkedly Concurrent wlth this dlQp in pH ilaS II fell inbull
bullbull
9
oxygen Such t~ral and $PiItial va11ations in ~andpft
would certainly qive 11se to corrosion cells middotatmiddot the sUbsttatie
surface ~ly resulting in iorroaien pitting
The r] _tabellc ctivlUee or fau)Jng -=raJrwerttmlbullbull
together with their prapeN1ty to-=ci-1tritla~
tlwlsalves wIll also lIied to 1dcIoenv~ lIOdifktIona
Resplretory activities wU1 tend to ~~ tn-the tft and Input caapanants auch a cinia~nY qatk
1mIertabJatesmiddot ara mlk 1bullbullbull middot the bulk of their nlUagllnoua
wastes ~ excrated as ~ 1heexpftunt ncord~ pDz
end pH cningS amN 1nd1v1Q8l~ ~ ~ri
claady 1nd1cat41 the ~ of _tft~t GIl tt ~ of lnIirollliental changes Inmiddot expr_~ta lllhere no Wlter
--nts occurradthmi a np1d dacreUfi1ll pDz
COIJIdalo be a ~decreae in pH (no a) v fladrlg
condlt1C1ns are r-ed tIwn Ieymiddotmiddotor iIay nat be a 1bIIeIqI1int PH
1nctease dBpwIding ~ the --- ofmiddot iheltr ~ by the
organl1n the cluIIp 0wIgn wui a influenced by the
proxJa1ty of 1nhalent or ~a~ the dlnsity IIId the
tIlJcknes$ of the clullp w111alsa t)IMt wiuence in ~
wetr -N_tbull Hencein thick clampllP aUt and detritus
cCUlllulatlon is are COIIIIiIn ~ theh billa IIIdfbls
restricted Br8IJIIltdow1 gf organtc datrital _ter1als CI3IIIIUIIS
oxygen resulting in recLlcing conii1Uons llleasurlllenta haw
shown that sulphids qulckly build up In such deposits and
sulphate recLlcing bacteria y tie-~ SRBconcentrtlonmiddotin
J -I
II II IJI Ii ill
II
-bull I
11
10
the depOSlts woUld terid to Ile lOW at We start of ~r1ments as
condit1onswere nat favourable (POstgate 1979) The cantlooQUs
prockIctiPn of r1th f~plieUdofeacal_te1alor IlUlpllonill1l
~ts bYpoundilt8~ (~~Iick1ngand 1IkIod1) hOweva ~ t H 1 (
IIIOUld rep1dly altAIr theli11l~t and the rua~ofSRB re
notedtofncreasemiddotttcllll ( 1 SRaIll1taat llaUt 1 x 101 SMIIl-1
aftet bull -th
IUctotJlIIl corrosion canfe=t _ta1sln a1Ullberof Ways
(Iverson1972 fUlllIt end King 1975f 1Uilr 1981) pH can
aff~ the ectivity of the 1IIin organlasinuolved the H 1IIho
hava tin qlu pH range beban 10 12 (Graff 1981)
although tIdS -can be lnf1u8nCed by ather fllltors In atI1tl11n
they lIKIJlre a catbon end nitrogen aurCa 11 various
salts tin alactron accaptor (HedJart ~ idbullbull 1985)bullAil of
U- facton are availabll in the debris -alltting b41tween
ar1na foullng -d the 1lUbstra~ 110 providing bull potantlally
corrosiveanvirorwent
Evan IIIhete such accullIUlatiori does not take place varlable
changas inoxYQ8ll and pH ~trations acrossthe suface of
teel soch as C3I becaustd by foul~ ~ C3I give ti_
to corms1an calla lalildlng to piftlng corrosion
There is an initial tllld8llCY for the pH to rise as a relUlt of
the excretoryactivi1i~ of fouling aniIIal and this 1amp UIIllkely
to give rise to corros1an problems Eventually ~er feacal
h
1
~I
materials and detrltus~ill build up around or~anisms ~re bo~
low pH and p02 devul9ls providing a potentially coJJOampiIu
envinx-nt flaw does dify thh _build UfJ but inmiddotthbull
Ixparlunts carrllldotttid notprlvl(lt it R~~r it
l8Iilated the depth of deposit ~1~~ in t- ~ It ahoutd alao r~~- in f~ population so-1QIJvlWela 111111 be dying or dUd Thlb
~lng~wUl~1Ielacal1Md conoa1ve ~iccandltiorJs-t SR8a 111111 b4I fCllllfC314lng ~y pOIi1desa hlbitt~ HtInd 1tt~ WIi~ul create 1~1Mct Jrcx-ntal dlffarenca ~ ~th l1kel1haOd or pitting
n
References
Albilnder J~T 1 bL~1953~~(9EthII prockIctiPn of IdcroaltJodIIs UIIId in I~ studies Rev Sc1 lnstra ~ 528-531
_r1can Ptbl1c Health laSoc1ation 1975 5taIIdIrd~for the lIIIIIlystS of _tars end taatms 11lth pounddIt1on 1IPHIi-~ 1015 ~taenthSt NW 1IIa-l~~OC 2IJ]J6 USA
Bau-8laquo=k1ng C~IIIG amp 1IIoOd EJF1955 Biologlcal ~ in theestuarlna envi~t 1 EcolClil) of the ~ cycle Kcnlridlkjl Ned Akad -t ~ Ser B~ fi 160-181
cter DB amp SUver IA 1961 In IRafaranctlElectrodl$l EdltolS Iva DlG and l-z Gl lc8dilll1c Pr London Maw Yodlt Chapter 11 ~
Davies PIII I 8r1ric middotF 1942JII1c1oel4iitrodas for-aln local oxygen tension in n11 tissues Rlv Sci InStruII Yo 524-533
EdyileariRJGJ ~ Tetty LA UJ83 The 1nfluenc ~lIlcro algae on corrosion ofstructural steels used 4l~ the 1tIirth
Sea 81odeterioratian sect 336-347
bull bull
~
bull - -- ~It Jf bull
- ~ 13
12
Terry LA I~Ed)VeangtRfiJ 1981 lIi(toal~IOI1CGrmianF~ l)i 1St1oJhemailnEifooiiiigffixed gftshoreIt Bot 1Iar ~ 117-183 lsect~~jlaUoQ1l1 _ ~t Ef)e~gy Offs~r1I Te~lOtjjy Paj)8r
-middotltcntpI)~it$t~l91-16PP~middotmiddotdl ( I
bullbull
iIIalk e1 JL 1971 bull Ion specfie Uquid I~~ e)(charig~ ndcroelactroclels Anal ChIIaI ~8S-Q2 AGraf1~ w~S 9S1~ 1orrosloh InFIntroooctI00 tooffshOle
51~weSoes~~fab~lcat~onIns~allat1on Gulf Wall PI aelUnger EG 1982 The appl1~tlon 9fMl1ikIngmiddotHoust(jfW~Q23g-258middot icroelectrade in the study of pH end PD2 in lIHinli
HerbierC SN fGH~rt~ )RI -Stoctcdalpii Watltif1Sal RS ~1ronIIInb Ub~~ 27l-Z79 1985 Fac~or lontro1l11)9 tha activity ofmiddot sulpMte
bull Wall P 1983 The appllcatlon Md ~ ofiwrJ)e~c~odl shy-taicfii9 tiiCtetili tn~ diiilncj tar infection
techniques for the iiaurnt cif icraeov1ror1liltital5QiCiety ofJebo1- Engi~ SPE 139781-10 _i~~ - ~ ~ i ( _ IKIdif1catlons caued by aGII8 rlnefou~lng -C~ebullbull
PhD lhaaia Ibiverslty or ~tar UKbull bullbull bull -
IIH1nke JA 19fiSa In EXperllRants in Physiology and middotampioClIiIiiilatJift l c E(If(orltGAiKetkutgt ~c-Presa-
Wooltdngton AObullbull Davenport J 1983 pH lind IlIl2 IMla ffl~- j~t~5 -lMIIleath 1IH1na ro-fauUngGqWl J bull Expbull 1
B101 Earlsectsect 113-124 Klnke JAIII 1969b II) Glass IUcroelectJQdes poundx1tors Uwal1~Ui~ OrOllt arid~t Nt JOhn fII11ey YOI1lt thaptar 11
~-f _f~ ~ ~~nt ~ Igt
bull Iverson IIIP 1972 Biolog~ Qrroaian In Advencesin
corrosion scianca and tecIYIology 2EdS FCntaia G~ I bull IIS~et RIIIbull p~ PJaSsLancIoIl 1-42
bull
Jorgensen BB Reusbech NP I Cohen T 19B3 Photosynthesis and strUcture of bIIn~ II1crobWp MC1OIIlectrocki and ~ stud1ea of four cyanabacledalt -~usllWtOl ~bullbull 1~f093
t l~ O~ ~~~ H~ ~9s7~c-gtHatste~iecfn1k van shy pellediftlrtenPt - Stlchelektraden (1-Su
AiJBitiKlificfiliilislflilr)zlIr polrapusdiailJIIissui1g_ liIserstoff(jruQkas fur d1eB lIsUbullbullung der IIliJlttOUf1cillatlon PflftgaHArdI ~fl39
bullbull Mller JOA 1981 tals In EconcinI1c II1croblolooy sect McNbUdbicldatariOtationEd Rosebull A middotAcadelc Praulondcn149-202
t Mller J DAI King RA 1975 8io$~iorat1on of lllltals In lIIi~1al ClflPlCts of the deterloration of tarWbullbull EdsGilbertRJ~ middotloveloCk 0111 bullAcadeic- Presst IILandon 8Jo103
bullt
IPotgate JR 1976 The sulphate-reducingbacteda
Callbridge tIn1versity Press Cliilbrldg8U-K
Reusbecb fUI i Ward DIII 1964 i~~~1Ida5tud1el of
bullt inter$Utilll IIIItez dttm1stry and photosynthetk activity
in ilflaI spring Idcrob1altAppl~EIi~ MerobW tsect4IlU-27lt I
IIbullt bullt
bullbull Ibullt
1
Inmiddot middotmiddot11middotmiddot f~~ a~ -tmiddot bull J~f
I~ W
i5iImiddot J~a t
ilmiddotmiddotmiddot I middotIt Ii
l
~
middotmiddot~~middotlL= g r
Lj Ic
t
i ~
I
gi II cEi ~ I I
1 t rW
D
tR bull
bull t 10
11
r
i
I ~
j
~
I
~bullbullbullbullbullbullbullbullbullbullbull
LV
FigS pH and p02 profiles into macrofoltling eomlllllllitilS dominated by Balm trenatU$ on steel plate 11 two pH profiles Ib) two lIOai profiles leI pH and P02 profiles into dead barllllCle shiella withdetri~Us I I
itIbl r p02 ~ I
nvnHg 11 70
j ifJi eo 1amp0
laquof it 30 s
I 20 N
ca 11 10 a o -- shy i bull
11
80
19
781 o 2 6 8 10 12 0 2 6 amiddot 10 12 dIpdI Qf ~iftto fouling In HighIst numbIII pIatIi
8deg~~
~i c5 iioo5 fiil
pH 8 bull_ ~
~~ ~
_ bullbullbull- pH
-pOz
(cJ
e-Clf ( j~JJlaquo1- --~I ~a ilil - _ I i ]1
~ ri en i ~ oJt ~iJ id tUI
2u1U I shyI t
shy1 2 3 6 6 7 8 9 10 11 12 i shyDepth 9f electrode (mm) into fouling
1 i
1
~----------~-------
~
~-~ lt1~
middotmiddotPmiddot ~
I
V
~
ill
1 Tbellitt~~ of cathodiC protection OO~~lIic~tlO aridrfOflllUlc
ofi AA~Uoullr14l ~1~I
I
ael~lner EG middotIUtcb8l1AJ Dept lt4 lInY1roMe1lal Bl01011
=~~venilt7 IlariCheW 1f139PL $ ~11al1df
r
I
Ablitrict ~~~ e~l billttI ~ted with ~pert1n ant11_1111ampp paint llere itpoii~1n _vatI- In the tl~ld iUIit 1bOioatbrt~ScM Were
1IUbjll~ to qtbocJ~C ~teltt1on byen _ current (~)SJi inci EDAX Iieamp~ta IJdlctecI ~ lachln teoc copper OIl tbePlCtec~ pa1n~ atter14 dQ1a~ protected Pllnt abo ebowed
) reduCed bacteiial activIty butmiddot ~e4 dltoe Iu1d protOzoanmiddot co~~l~tlont~1l IlU l~byenCOCltact not byen dUfwIl00 ~r _baRl lrillolvedln tOXln nil ereio noted
l ~~
1
I
Introduction
Applications of protective coatings and cathodic protect1on(cp~l
are recognised corrosion control methods for IIIBrine steel Itructureso Cathod~c protectlOll preventl conCIsion by Int~lng til8ctrl~l
currentS trem externalllClUfCa to e~~tthlJno~ e~ carrollon feetlans C04U lUChu tUauJ1ng Aelnts a
(
barrIer to ~ the flao of couos1a1 ~aboIHnu~r
Dccurrlng anodes cathodes or wlwit gelvWc CCIIII1 ( 19681
flinger 1980) AnUfauling ~ also JJICOqIOrII~ b1oc~~
Tr~ylt1n nuor~ (~) end o~ Oxide (~) (~ flJ1J
- of u blacld1111~
V _Ut Evans 1981) ~ ~
end growth of bipfoullng 0I1I)IIIII1IIIIe Wdd~ ca ~ of
paint rusendmiddotmanemiddot ~(r~ til )
As corrosian ~t1on ~ bICCIIIII -an ~1tlQd ~ of
antiroullnQs 1 conjunction _ith CP have bllClIOr ~
resultIng in reseudt 1rwestllj111tlng trw COIIIPIitlbUlty or VU~ ~t t _~
rOratlation to dlfrert levI ot CP (~Eelf 18110 ~
~t 1980) RelatIvely 1nIestiljlllUans have ~ urtw ~
or CP an thIt anUraullngpert~ of palnta
In thIt preseot 1nlIestioaUon tV~iln111 ronutlon ud
cOIIIHrclallY incanjlntlon wIth CP was ~t4IC~ Iw I sraed current (-966 l1li) arid exposed at the 1 Strait IInQlbullbullbully
~t1ons weremiddot 00 the IIhcIrt bnifects the PprIUd_
had an a) the various COIIIPQI1eIIts of n pe1ntf1l11 end b) l1oft~
developaentbull
bull bull ~
~ ~ - f
bull ~ _T~~-middot~ ~~- ~r _ -$1
(1) ~rlttionor Painted Steel Billets middot_ ~~tmiddot_tlJ middot~hmiddot~--~fmiddot ~ middot~A~-~~middot - -~ ~
Thirty-al ateel blllets (8-10aD2 end l depth) were cut frollbullt ~middot~middot~middotmiddot~middottrSmiddott ~jS-~ It bull gt~ tfl - middot--imiddotmiddot ~ t1dItrd steal plates The dimenSlonsof -thB butits-lim chosento
bull -~~ middotmiddotgt2h ~ ~-Imiddot ~~rt ~- -middoti~~ ni ~~middot1 l middot-~t~~middot _
(a) fecilltate amiddot Ie detrucUve -thxI of prePraUonpdor to ~~_gt gt~ k7i ~_ t~ 1--1- _ ) - +--_~ -_~bull ming electron ~ 4II1CI (iialloWdirectobs8iVatlCnS of the
- i lt-- ~ gtbull -o1e billet 8Uface
gt r~middotmiddotj ~L(~middot~middot rmiddot tmiddot J ~ - ~ - lt _ bull Single non-standard (1hIula~) nickel lIIireSmiddot Spot Welded e1Cing
bull ~1deJ ~ b~i~ ~j~~ tri afl~1b1ipOi~ f ~1t1 ~4middotmiddot f-gt -~ ~S-- ( -- - - _bull resin EactI billet then Undb~ tI1d prtiladlllltha5O __ eyilr
bull ~~_ ~~imiddot ~1iil h~ intlfcili~ ~eonbt1nlyi Tr1butylt1n F~id(18n)~~~O~dettu20i~blcicld8is 1IIfii bull i - ~ ( then 1IPI)l1ed to glvaadry Ulil tIi1c1ltnOss or 1(1)-150bullbull
tbull (2) IipreSSeCi Current Syat
bull A rigid ~tic due 1I8S drWedllliihwrtv-s41 liiIihOieS IIPProX~
bullbull 05-10 cia episrt ~ Pilnted ampUl8t Wsfbed IntaPQSIUon ampy
threadlng the nickel IIIlr8throu11 the dinto a tarproot junction
box Eliltilen of th8~1l1ii (recelving ~t-d curnritlere IIIlred
Into bull block connectOrrecelvlng dc trOll is potentiOstat The
rainJng iontroi blUets Wet8fbedfl11i1iiy lIIithln thejtlytlonbox
i~ityd~c IkiliPly ~~ ~lat1ruaWiode 8ncIacalOliel
refer~ electtodetoiere fllied ~ the per1aetat of the plut1cdlc IIIId
connected to the pot8ritlostat The desl~ of the potentlosbt ailoiolEld
~PlDteItlO1TClJrnnt to be MIllntained at 8prampset levtl of 966 II1II
c
r~
Grid raf 1J5s-t 114iH 563724 dud 0 Juna
Alolecule andbechr1a in general are ~ to
sutface lI1eroleyer (nlUSton) of _ter (~bullbull 191KJ)~To avold
contaJnatlonof ttw painted surf by U- -vents
blllets Were s-nact in a 1 conte1ner or atc1le (02i ) f~~nd
ter Iih1ch then f9ICI in tv 1IIhere the b~
be r-oved undeNeter El~~ bllleta fa Mcb trtai bullbullel
removed over a 1 dtq (336 hr~ expoeure perioda
each being ~~bn uposurnof 1 5 11 iA 168and DlIIauz1r
Arter field axpauHli the billets wan
fUtered _tar to ~ 1_ debtlflMd far 10 aJnuta ~
wv ~~~endattid sdbUiUd(otO
~ ~l f~ nn1ng el~~ --
lOOted onto 1middotCIi ~bullbullWIsend carbonmiddotcoa~nmiddotmiddot~middot f
each billet Ws~ Iis1ng a ccr1dge ~el~~
(5360) IIIlth anergy dJ8perslve X-ray alcroenelre1a (mx)
_lysis the les Wen reNed end coated III1th vold to al]Ql En
detailed sat of the alaquofa1l
(4) Lfiborattux ~
Painted taa~ bUletalllllre pnparad ~
in a Mrlne reclrculet1ng flOlll _ ~
f1Oll the ~ ~a1t~y Tha~~Un bullybullbullbull
10C M1nta1ned ~~t1as of ~t~la protazOllbull~bull10shy
(Entermap) ~ ~~~middotNlt~Eamp1~ __~and ~ r - ~ -
certaJn ~lc 1IQ1~ 1(1 the
the painted
~
thre repl1catft fn
plKed 1nto(022_)
- thIine1J~~ -lt bull
Atter mAX
(1-2) and i-n-t bn collected
14OC
Pain~
-
12 daY~ exposure and prepared as in d)~ - t
~I ~~~-- i~yen 1~ v
Rflltsi ~ _ ~f-~lI ft j bull _
rt~ll0~~~llY~~~~peln~stetsl~11i~ex~sed for
y~t1~J~ ~~~~1~1~~1UII~ya~kooJ~~~lt ~ ~~_~~I1J~ ~~~tomAX -lysis ~era than gold ~ bull bullbullbull c - _ bull lt
~~fllr ~~~ vllUll ~t1ans of the rfee htures and gt Co
~f~ - ~~~ ~f ~ ~aobta1Mdue IjIll811 iI1Flg 1~2 The IIIIdn ~~ ~ lt)ji9 )tmiddotmiddot - ht
- bull
~I rIIplt~ tIt8 -~ cOlllponanta of the pei1t 1bullbull WelIIlItr1x $ilj~t-~~middot - ~ _ -_
-IUch PVCbQed end rap~13y the chlorlna peak the
i~~~-~~-~V~~~-t18 ~~~~ ~~ Th8
ral~~i~ JlrQP(lrUo1sof ch 1Il-ntal paek~ll1d l~t_ Q~7 -gt- ~ _~ -- gt ) gt -_ -lt ~~ the 0 -end _ xprtsntabullTIwsa changes are -r1sed ~t~ lt~ _ ~ 1nT~ IThe8llia$lb npresentthe chengelI 1n~t1V8 p~lons
ti~~~ [ ~gt 3 - - - _ - _ -
Qf nellllllllt within We surface layars of the pe1ntmiddot ru The
~f~~thtJn -d ~rfeli avr the 336 h11cxI ampScanbe
SII4iIl r~ fig 3 al~ tht rta of~ dirfers tJetwaeo thaa ltmiddot~1- 1ft~- ~ _
IJOl appfiai-ed to ~1nlt1elly until ftar day 168 it fall
~~l~ 1h8relllllrelgn1fi~ diff~be~ qrOtactlid and
proieCted (at -966 ) piIIllnts tor lioo COPPer but not folimiddottin
Ibi~tlons Of the Pain -urface 8hoWad _Pil ~ witilt1M
(Plete I to IV)~ 1hePllat1na (~) pelnt(Phta x and b)
Clearl~st-i~~r alots on the SurfllC8 IiII1kh~ter1sed ~- f hlt1 lriitUl tin conCantrat1on~ lllariy slotsre Partially
~OIIe~ ~ tile _tr~tedal middot1lere rs ~an~ densely packed
grerltr~ ~th 1ar~(iIa1niy cotifIIr) 8ntt -ampniBil (lIB1rily 1rori)being
middot vl~l~~~~~ Dtt~i~~~t~ r~ ~f~~ I~t seawatllr for 11 n IIIOre slots appeared and th~ ~~ess _trb cwer
--if -- V~ c~~ f(i ~~~i~
(Plate Ie to It)~ the granules dld not protrude from the~rtace in ~ ~~~~j1~r~-ffJmiddot0~~
suhap~~r Both 0 nd --960 at ~~pa1nts ~ - fr ~~~~-w1-U~ ~~t
sl~llrat thi stag~ (Plates Ic-d II1d xrnspac~IlYh middotmiddotWith middot middotaJ~cmiddot gt incraallO ~alolre t~ difrrqcbullbull middotbatIiI8IImiddot the protected fjd
bull ~ ~ ~t~
IIllrotacbldpaint ~~lIfIt~ftitr~ h alQt~ p~t bull u bullbullbull J ~~i~~ 1~~
lIIith IIIuper lIdguon u 4lrotatad piIJnt (Pl8ti1 II) 1hD1II8S
Ill )1IMdw1lillS gnnJla~on f-r~~ dpert1cl ~ - - - ~~r ~t
Onthe 966 1IIpetnt t-var tIIO typeaofan with diffannt c t
~hIId~(Plate UI a to c) One hIId tIII-1 bull bullbull e _ bull
fUll wJ1hleas abvlaus alats the o1fwl bed C18UU1IlOt ~litUi middotC c ~ ~
less ~ t11 h Gnnulatlan __ but ~lIiIZly def1nlld in
middot~ latter Afte fila bed tiIIrtIId 10 ~are ~~j(
extansll(tlly ~ the~tIId(O at) tIIIIple lIIIdngthe~ - ~
middot orarallet1ai leas claaI (Plate I~) f u II1ao CQ1tJAJ11d to ~Gn - -
the -966 at 1 (Plata III d tor but 1JI8V8n 1nittNdcneabull -
Althatqa -middotslots wrs abvlaus becaI1ng 1 cwampr aiI- the orwWat1an8
f1nailyftar 14 day exposure ~ fUll drMIlapIIIent on all la - ~
are extensive with the 0 lhawlng both alota pore ~
OVIKgrOloln (Plata II c to f) Gnrulatlon 1bl to ~ gt~t~~ i
or the th1cknats of tho ma ~the ~ at ~ (~~~~ themiddot - - _ middott ~ ~ ~ - -1 1 t
middot f11bull IIIIIS IIOre extans1 thIIIn at dIlya but IIIIIIIY bull1Q~ tI st11l ~w -middotr~ 1gt ~1 ~
obvious fIIany ~ loI8re still p~ yen~~~~~~~r
than before Poiu appearedto be f~lf~ila~~c-~~~a (Plate IVf~ label 1) IIIIl1ch broke~i(2)~ ~~~~
t t
bull ~lS (3) ~ rflC~~ ~i~t~~l~~KI~ t~ldiIiiicyul~loIiih ~~Rht ~ )~- -~ - ~middotd 40 ( gt ~gt ~d lt1 t ~ Jt ~remiddot4amp tha paint trix cavity bottom eroded away reVe81~ng ore
t ~~t~ por~~ ~l~ P~]Y p~~8dby di~lutiaA~ ~~~~ti~Jr~gt~ta(l~~Siinll1cl1Ud~f iUiIiI reriatlGllbullt
bull a~~ ba~tid colCriAtloriccic~ GIl ~~~d ri -966 - ib~~~amp catQn1iti~~t~I~~(~t an ( ~
t the ~~~palnts (PIa_taiiI d~~h ~wh ~ n td have ~ CamplluJMitiIVc) ~~middottti o~lgtnalrf tlt of hIght
~ l=~A ~J~rW tYtles~ p~fiiilriQcOCCold to~Ja to jU-t ~jmiddoti~ai dlatClilS ~ir -jlati ~tbullbullent aNit ~ t ~~ th8middotf~ld~ies~~t~on~ ~ -966 prQ~~~(P14ta Iii~)Omiddotiliof~m1hiiHAiiipieat ere~ll-(piitir~ p-iot~iilans(PiA-iII~ end c) nbullbulltiledt dl~by1i~~ end wirinali1sheii ~cMI~bY thI tili
bull In t-~~ ~1i1nt 1oIIIeHfd~ ~m1IS (15OClind a t ri~r-dlatoa ~lor P~t dlataiOlonlt1anGIl ttw-966
t
bull ~
pr~t~ ~is~ih iion obitJauIJ A ~rot -=1 ~ ~ i __
reprasenttild on the 966 painted 1iUrfc1ncluicunljj ~ rwt
Achnanths Colonisation by bacteria 1n the laboretory bsed
bull eliperlMnt tollMct sbiUar pattern to the fild ~
bull bull Dilicueii
bull bull DUt 1Iidnt ~lona Mve dlttllrint ~~ ictian
It ~ JntM l1tenture ttt Iff yen __ JIll the
bull ~ bicins ani 111 ~t INchlnil (PhillIPs 1t73t EWene~ 1981
In ci1ntlHt tJWY ~tit ~Wl1c iIuCh as 1rbiltyl tirI
~ lite wAlII]ltiClr dlsP8tfu~ middotthiirsttilll~intaJn~ a
bull ~
lIIifoflll ampalCentraUon through the filII 10 thoIt biocide rel_ at tha
paint surfce 1fI$Jlts in IIIOre ditfusing ~s to repla~lj -iflJ~it
resulting 111 bullbulltNdy lr leechlng rata M1llat this IIIIY be middottruIi ot
tin ~ it was nOt true ~ ttw middotlBlf fOZWJlatiGn ua ~ bull - i - bull 1(1
ObSfvt1onundr SE clarly ~ tM t1l1 to be present
~ly lIS OIIIdle-llk parUc1~~t tn lUlfiice (Ito
IV) bUt usa ~ the dIpth attt fU-middot ~y~
If one considra the concn~rtlonmiddot chMge 111 1n ~111c COKICIl8rlts fro- the paint tU- i bullbullbull tMWei b~ ~ Si6t arid
tI-~t iron oxidt 1~ can bt (Fl_ 3 IrId TIibl8I) ~t ~ ase cona1derllble d1fftrenC8s ~ a Un decz bcdYMr ttw p1iod IIIIwraS lron~- an unt Ught inCr h ~ incqJper ~tlGn occUrnd _idly at ttw ~~ Gfb ~ period ~ out ttwn fu nPldly ampQII1n ~ umiddotirid II phast This IIIOUld bt tIlq)8Ctad It cantact l-=h1ng we occun1ng
IIltfIOS8d pertlcl of copper would lMch rapldly on t-n1on 1n - shy JIsewtr There would thlll1 ba bull J-a a1iltUy leu avalUba
i1 ~ ~
_tedal becfM uposed Thi tIOUld ttwn IIllCh Thla pulSing efttct
IIIOUld be expctad to -oth OIItlllith U- putic1 __ -lllible II tot 1~ GIl a on r-x _lao Thla teet 01 put1c1e 1tUng
can tit GIl the lIUrlace ClIlC8 it ~ a pan 1amp t
a 11aM bet in Plat I to IV IIaIrIg _ lit la_ II ~ 0- ~s _- PIate IW $llldl ~bull nshyot poRI flllNltttn Al~ tin ~ lou rete thtn I bullbullbull atill Ivwnce of bull al~t pulsed 1Itue rrlict tII1IIWd
elqlCtaG inIl ttwauUr ~ I I I
0
~ i
~ ~
~I J
bullbullt bullbullbullbullt bull bull ~
bull bull
I
bull t
~~
The same seqiHllle of cevity follliltloii (intttecaseofthetestjjltle
slot- shaped) fClliOileciby SecOlld8rYtlolesfotlllingin thll cavity
bottollis releasing further deep8rtox1ils (Plate XV d and g) As
cavities_re ProdICed ~ tiecaaIelnvadedby_ter~ onelilauld
expect 1ncreaa 1n1IOdha and dIladne lMls t the t1met
which the toxlM uw ~ Ths fCMld tCtHi eo (Flvmiddot 3)
~rttno the vi that_tat ~t1on otthe~ ~lY
il1Cl1l8S8S Idth UIiJ
Differences f~in toxin li~~to Ini 966 ill 8811P~ F ~ bOth Un end ~ lcMIr levels afillW
IIIIIlfI ptaInt(~1~~1ntha~~ ~~f~~ cst rklld with tbiIccpper ElJIc~l ta~Uona invalvinv
w pl8cId at a~ht~~middot3lO~lth Un cqJpel bull
(-960)10 ttW ~af this expei1tIIri1ts thIsiorelfJltdt -
that cqlp8r b8IiIId ~ IIiIOUldProc-ln f41ip1dly t thellllgllitiv
patentW _lied () as 1t 11 further f~1t ~ witt Un
III11ch is u-tat aqulllbd Idth
it If ttda 18 eo~ ~
~ )
frOll1 paintscould bl anqnced by an illprd current clithodic
protecUansysteipos1bly ahart_nino te 11fe of the paInts
antifouling ability Bacterial coloni Uon IIIU rtdUCed on the -s66 paint This
could have been due to unflSllOUlllble electrostatic cqrge on th surtece Th1a effect CIIl pley en IIIportant role in the t~of
bacteria to alrraces (lIIarsMll19E1O) As bacteria appear to IhiJw
greater toxicltyto copper (DMpsey 1981) it is Klre likely that
enhanced cqJper leacll1nl On the -966 AIIIPl c8used the recluCed
activity The IDlol8r toxicity ofUn to bacteria (as reported by
~ -
DelIIPsey Hle1gt or its lowes leaching rat was sUpPorted bythl
relatively rapid recolonisation of the slots by lIIicroorganiSlllS ~ftar thE inlt1al aurge of leachi~ Thailllpressecl current dto ~~ the oppoSite _ff~middot ~r on dtta colonstJon l1lI-1- in the
JIbcJrtory~ta larger rUDIIra f~ fit pro~td ~ther than uriprotected Pafnta
Ccnlus1one
1 tiild-a 18 presaIIUd ~ thattJn~ ~~
~llNChng notd1rf~ ~
2 TIleuseofCP~ ~CUIftnt(at) on1h1a ~fca41ng ~- ~
pa1I]t~ to ~_lAIIach1ngn_ at toxiN - -
3 leIJstaPidbectar1al ca1on1aatJon acaampmId an ~bId Pt1ntJ ~
4 JItore larger alcrofauJing (Oiat_ and PzOtoaoana) Gr~_
occurred on the protcbld~
I l
I
bull bull ~
mfEflpoundaS middot~-t9i _jgt- ~ _ _ ~sev M~J~ (1981) (cilonJzatiooof alUf~lng lnts by _rine
Jbull ~teda9g~1ta IIarina XXlV~ 185-191
- - gt 0 Ie 1 _ - _
poundV8I$i I~lf (1~P~h 1II8rin~Ugae and Foul lng AReviw ith
t p~l-i9Jrlr ~iiMdilto5hiP-fQUllfCiH 8ot~ ~r~ ~~~ XXIVbull lIPwl~J11A
Fotteth CN~R ~kk8nGB IWPIR1seJCiiiroaic1lihd ~e ~JIIth CII Offshore St1ucturu ElllS ~tjI~f~1Dt19i4
- - ~
UKC (1976) I~races ill ~W Ecology KIuverd Univbullbull middotraabullf~iddge ~bull C~~jII illt)Ia(t RC (1980) totinllsa Cattod1CPrOtect1cinbull middotmiddot8AtEmiddot~6haOCIlfif~ T~ p No 108 ~ 18 ~ 1IIHf bull
t bull lt ~J~~~ __ ~
Norkr4irulB(198D) Advanculn UcrOfj~lpound~ltiOyVol 4 (AlItxlaquoIder fl Ed) PlerIw Press lcxIdon~ lIP~ 51-85 ~ l~l - bull bull
PJijllt~i At(fm)~mr~~JnI~1fqul1nQ JlalntlJ Riumh ~JnOrpnic toatJngs 2 192middot
)~J gt0 c ~lt -- ~ __ RogerS~ Ut (1968) ina ~(lnt~fbItliP4ili GI COnoslon ~a Technology)bull bull
Sl~iorr~ VP~Ranald~ Rctd~~ (1S80)_ ~ Experllattal Stud1aiRelate middotEffect or Cathod1c Protict1Oi withmiddot terta1nr1c
Coating Syst~ 12th Annualofr T8Id1na1ogy Co1fItHnC8 liauston TIIlt p 9-12
j
fIG1 sPct FromMon IT8TJCuZOI Int Lliu EOAX
FlG~ Fro TlTCu20Ar-4 Dy Field Ellposult I~ir
il
I c
R hi -00amp=7
-I I I I I I I I I f
iii
bull bull
bullbullbull
I J
J
I Figur 3 Thl chan In reltl rqpoIQnlof -ariOtimiddotjOmponents of the paint film with time Thl vtic~1 ampkiS unl till _n ofmiddotmt Illment per unit OJ fe 50all Cl Na
Cu StI Feeshy
PlistinllFe S7~J6 704 3~74 908
f
r e ~ ee~
middotie ~lt - ~ shybullt bull ~
t -~ l Cu~ o ~ f
t --- 3igt - bull 0 J Sn
cc
t ~
~
bullo
~ bull e
bullbullbullbull bull ~
bull ebull a QI 1- bull 0 40 431282 316
101 dmllIII)bullt bullbullbullt
hr 0 ~~bulla 5~16 ~~ -~~ i166 4S i83 8~1B
24tu 6~480 SD~83
111middot 1~1 OS6
~ 5114 6~69 -51 UIamp middot016- lt ~rmiddot
1d o Mea 818 21 021
~Z21 199 1~1Qmiddot ~Jgt2 058
14 d o ~~ 804 Q~B8~~~a~~t~middot 685
866 372amp 8amp4 OS _i~11[ ~~64
Tttile 1 A81UvlL pzQJJortlcw 0 vxl~oI ~~bull Jn 1I1Dtitctel lt-1166) end lqIlDteCt6 (O)middot~af~_~middot_lodEDf~ middotmiddotmiddotmiddotmiddotmiddotmiddotmiddotmiddotc
~ ~
_----I~
J
I bull
I
J bull tmiddot
7o CONQRESOIMTERNACIONAL
I D~CORRO$IOr(IAAINAEmiddot~USTACIONES~v c)
t 7thflfERHATlONAL CONGRESS
ON IARINE CORROSION AND fO(JUNG
~CONGRE$INtERNATIONAL DE~CORR~ON ~E ET QES~SAUSSURE$
i t
~ ~
bullt
t t
UNIVERSIDAD PourtcNlCAVollncio7J1 NiwiemIif~ 1988
(SPAN
SECcl6lfn
BiOIogro marino Morine biologY ~1i1orine
Theunpub1ishedJllBIlu8criptEl have been reprod~d asr~ielved from the
authors The SCientiUc COlIIIBittee take not respoi~ulbll~~Y for any shy
error or omssi9n
Laa ponenciasno han side publicadas y estanreproducidas tal y como
sellan recibido demiddot BUSmiddot autorea pound1 Conte Cientfico middotno se responsemiddot
bil1za de cualquier error ~ om1si~n
bullbull _ bullbull_ bullbull ___ bull $_ _ L _ - _ --- -__
SESSION II Marine biology
Friday 11th November
SURFACE PROPERTIES ON NONmiddotTOXIC ANTIFOULING PAINT FILM
K Nanishi M Murase Y Yonehara M Kishihara and T Hirama
MICROFOULING ON ALUMINIUM PANELS PLACED IN THE TROPICAL WATERS
N B Bhosle K Venkat and A B Wagh
EFFECT OF TRIBUTYLTIN OXIDE (TBTO) ON THE OVARIAN MATURATION OF THE PRAWN CARIDINA WEBERI
R Nagabhushanam B Indira R Sarojini and Mary Frances Thompson
EFFECT OF BIOACTIVE MATERIALS FROM SOFT CORALS ON BACTERIA ASSOCIATED WITH COMMON FOULER
Vitalina Mary Avelin Mary R Sarojini and R Nagabhushanam
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
t
f ~
_--
yshy-~~
-1shy~~(~
Jl -
- _JIJ 11 _ bull ~ bullbull L IIIJIIM
I bullbullbull shy
-
if Ilt
JII l3$i]J ~e-l~ ~ 1
J ill KItt
Ii I 1 8 Ij j iJ)Sj
emiddot Sllll0 r B 8Je Do Ji
t ~l u Ji ]
1 0
j
j
r 1l3iiiJail1h
bull III
~ i J ii i )1 t~I ~
~
~ )38 ttl i UN ~I3 11 I ~iii bullbull I fi~fi IelI M 6 6 bull s plj i I Jjhi
~l -u II11 i~ Ji ~aJ j~ ~ tll
- bull
-------------- -- - --- -- - -- shy
2
This paper desr1blS the use of ti1c~lctrooes to measure pHbull bull _ bullbull ~- l bull bull
iJQ2 chmgtS ~~ 1II1C~olones ~middotilIlg~ ~f~~s i~rsed ll ~ter ~ wlJlch t1)U11ncjotgaI11sllls are grolog SOllie p~tnt1al 11P1~~tions C1f~se cIIatlges to corrosloO at the
lt I )
surlP alllO br1lJfl)dl~ - 1 - ( _ ~ -
ItIiterWs 8nd ~ ~ ~
() poundI~~lt - -
Ttl lec~_ 1n theu bpelsnts hIM bnndeScdbed In
~~~I )iel~~bull~~i~) but n 11 ~ haJe
- Aj ~-
(1) pHel~
ThapHalec~ lII8rII basadQIJ tne-iWof It1tQ (1__)
wre the pHlt1V8recenton of~~l~ 1a fiI)OS8d bull 1Ihart po1nt extlndlngfro- tt lnsulatlnQ (Flljj1) The
pH andmiddot alUlllno~nlcate glau 1cropipettesued lnthe - - ~
electrode constnction went ~ using bull wrt1cal alectrode
puller assntielly bull vart~c81 version of the equiplllltnt
orlQlnally developed by Al8Xllllder and NasWlaquo1953) The ~ ~ -
electrolyte u-d1llllS 1N tel and filllnQ IIIIIS 8aoIpllshad using bull ~ - or
aydnge 8nd a vary fine draIIIn out Qlass capillary
Once f1lled the elfCttocJes were soaked for severalday~ in 1111
tel to allow coqlletehYdroi~s ofthe pH glass lin lnternd refelflf1(e of sllver lrecoated wi til sliver chlorld~~~s
ccinmctedmiddot to a gold plated pin iealed into the imd of the
electrode lII1th~ill~oo rubber COMPleted ei~trodei er~ stated Up downIIIards lna dilute solution of ~r~iIcrcj In allmiddot uper~ middot-temal reference Qf bull~~l
allCtradilllllS UIIIId DurlnQ -~ts -n bulllact~damiddot was c~cted to ind~ ~~F~Y(_ itrgtbull The cStcu1t layautforbotitthiiptl lIlIdpDz electtaifes
~
1a IIhIWi Sn Figure 2 bull
~ -~ - ~ -- - - - I -
CaltbnUariof all alacbodn id Clirr1ed middotOutmiddotbllfGn ftc AIIICPUIIIri inmiddot c- elctmdeiimiddotmiddotmiddot_middot~
ckalng en apars-nt by in nJIalt~ orthey dldriat
pEOCia ~lbblIt ~lntal callbraUon In ~
CIIIIi8S all ~ datafl(WtMI middotmiddotdl
(11) 0ItygIn a1acUocfes bull
TIUI catheter typa elachode use a fiat ended and of duw 1 to 15 _ It conslatad ofa ~ uia P~tJft- wlra
aeelad 1n 9l~ and IIIDadded in 1IPaKY~1o 1I1th a aUler wue KUng a raferance (see FiQ3) Ihen the epOxy hId~
h I of h l~middot careMJy ~ flat to UpQM
bath the plau _the ~lwr 1IIIIIbnre WIis ttn8llPlled
ecrosa ~ flat lind by rep8atedly d1fIPlng Jt into a aQ1utJDn Of
collodion 10 atheta1cohol A Pola1-1slng voltage of m all __
applied bit eachelectrodl and these re not thariuSed for
axper1lllental purposes until middotthe response I118sstllblallihen
bullp
4
bullit tuffer elut1on a~ a eorustllllt
it i)
~~ ~ ~eted IItld accepted for uSe elliCtlodes
it lt gt ~ ~~J ~J _ bull~ - - -gt bull
IIIIiIr~~Jn~IIOWt1on of 02 KCl D~ygen Ilectrodeser _iliry~ ~~)~ 5middot -1 ~ J~- - f -
it cli~rated aga1pst solution I of known oxygen content (as 1 w f - w ~-l _ bull bull
i IllllllaUred bY n of the l111nkler titration thad APHA 1915) ~Lmiddotmiddotlmiddot ~ ~
it Any el-=trodI not lIlCtIib1ting a ilnaar response rejected ~ ~ ~ ( _ - ~
zelia axyg8n lution obteined by the usa of a buffer
i aturated wUhpyrogallol crubbed nitrogen The 100$
~~~ reed1ng obtained by bubbling a1r thlOUltl the ~ ~~bull ~ ~ bullIII burfal far 12 hours at ~~ture 8M pressure- ~t bull
(b) [)cpars-ttal~
All -rs-tts IIIIIl1I Wried aut in atellperature controlled
bulli it
tank pliad an a teflain NYler ted l~ steel patfon ill insidil a ane _tre CIbI Faraday CIlIIIi(fig 4) Tbe~
prav1decI IIf1th a lJrfQII hinged door at the fiant for eccns and[f as gro~nded to earth to scrbullbullnout radio frequencyi intelfuwce The _tar flowing ttmluitI the experluntali - tri pullPed at a controlled rate fro a temperature
~ controlled reservoir For all experillanta involving algae
i llllA1nat1on provided using daylight fluorescent twes if l1d teel and 1Ial1ns plywood plate approxilaately 20 bull x 15
wire Ixposed in the -rine environmantfar several IIIQIIths to bull
allow a heal) growth of IIIICrofoul1ng organi_ to deVelop The
bullI steel plates had been placed utldally and IIIItre exclu8ively
bullt
~~==1$ and ~klOOrganl_ ~ldirig Itltena ~srar-nt of their
OWl The wooden plates 1IIIIIl8~sn the Jntett1ifalteclJQii
-f llllllre COIIered lIIIith ta 01 tt- alp InWfCld1IIPtth
occa1a1onal bryozaens There IIIIIIl8 IWIo lagII quantitibullbull of
~ retained -gat the algal fu-nt nPiate lIIIIIle alsO cleansed and stalJil1aad_ bllton ~
provided On 12haUrly~ ~dllrUght luar~w1 RddtUOlllll steel plat8s ~~In Alii ~dIxk~ lit
Uwrpaol (lie) IIIhare they IIIIIIl8 co~ by middot~Of~
(xtllus edulis) and the lISCld1M Clona intUn1l us
~1IIIIIl8 roved f the pJaqa NP~1id In cl_
ta tor 5 days at cansqnt ~ toIitDUampe
pH WId pOz ~1ane frc the extuMl t~~
the fouling to the Milstrata llllllre ncarded tt elecUadM
middotheld in a rigid pttllpex ~rr1et attechld to II ~ly
Jointed GIl on the lI1crcIIanipulAtor To nciad the muM the
tnraugh the foulJng tI1IIIIerds the ~ta JI_ta 0( 1 _
could be acCUllitaly controlled by talng the tIM clrlw of
the n1pJlator Iftn each lllc~ the DVIII ~oIdlft9 shy
allowed to _tabIU for ane wte prior to ncordJng the
VGltge~
~
bull a
-lt 1
l~to the_ batnacl~ 1ls~ COC)trol - fmiddotmiddotmiddot
recod4l9sof W pH 8I)d pOi at the SIJIIEdepth outside tOe 5hell middot-middotvmiddotmiddotmiddot_I~ lgt~middot middotmiddot~_middotmiddoti 1gt )
wete made
-flesolts ~
(a) pH IIIldIDz p~lllllS
The _re~ts for Hancl p02proflle tluough both algal -d - ~ gt Comiddotmiddot i -
eniIIIalfou11ng are gi~ in Figs Sto8bull --
TjpicalProtUeis daIiilttuougha t of Entronlorpha are ~1n -bull l J ltl ~~ -
Fig 5 - lIhllsttyp1cal d1innal fluctuations CICCUIJ8d during bull ~i bull bull e bull bull
photosynthetiC- II1d non-phOtinwnthet1c lods (Fig Sa) mad _~ d q _ _ __ _ i bull
decreases inpHlIIIIl1I~ IIIIa) fro-theaul-f1lCe of ttWt bullbull bull ~ bull
dcyenI tqwards the abstratacn tdch it enclOSlild (fig 511)
~fal1s1n pH--nted to as-ach as 2 units Over 9 IIIj
dIpth
figure Sa ahoid the thangea in pH -d Pl2 between 11ve barnacles
with depthln a stlat1c envirornent A steady decrease in ~
occurred as the plata IlUrfe~es toiereappro8ched pHWtially
fell on one profile before rislngbut lOSe steadily with daPth
on the second praflle SlIdlar changes are IIt10wI in fig 6b
with depth into the shells of dead barnacles 80th oxygen -d
pH levels were re~at1l1E1ly steadY ~ similar to the 8IIIblent - ~- lt
envlrOlWOt Until the -actual dead shell IIias entered When a
IIIUked drop in both fJ02 and pH CICQ11Tedbull
~llell in rigs rit JllellllUrellt$IIere maltle ~
iOlfillidual IlUs$els withtiRle (Fig 7dlb) In thll first
experisent elactrodes were placed indicated (fig)
arCUld indivicUllalsln genU flawJog ~ter The
resUlts show bull ~tle fIwrease ~ IID2 eMIS bull aiJc hour 1Ucj
The pH~t of the~ II4ttIn ~~atmiddot~
81 (te UlbientpH being 80) but 1GII8 ateIIfIU ~ the
1rtIa1ent aJphon to a6bull 111 iIadIIr ~~thI aff~ of
flOiW 00 local ~ciIndltJans~ the ~
~tadbut tile flow at~ turned Off tar 25i1aur
Jadin thelllddleofthlt ~The nailts g1R
in FlQIb -PD2 IIMtls mae ~y inflQldng CCnd1tJanamiddotbut
fell -zkadly fter alhoit lao w flaws caadawJnit a
~ fall lit 90 alii Levels rapidly ratond onca t IIIIUtchadDli agaJil No ILICh teed varlauon ~
in pH around the eldlalent 1pbn 1iIIflImI a taIdyJncn4IM -
obsmt8d aspec1elly cleM to thIIen1Ml CondltLni around
indivOJa1 ~ lIIhllst of interest are lMa repl_tative
of field conditions fro around groups or clIIIPS The
IlICpIIr1ants were ~ ~ted Wittdn denSe clUIIPs of ala
fouling a plate These r8SIJlts are given 10 figs tie (1Il1Ud
IlUlIS8ls) -d 8b (within byssal thrtiIds) In tatic ccinditJans
amprCUld the _15 u-a1(8$ ~ fellrapldly (45l) reelJ pft
rose sl1rjltly and then feU aftar 2 h llhen flowing condltiors
were resuraed after J hthe ~ rose to the original middotlevels shy
The pH rose lArlcedly at the eltictrodenear to the xhalent
bull bull bull bull bull
bull11
bullbullbullbull
it
I
8
s~ (3) but reriialned stlaadyahd ultilllateiy fell silghtiy (2)
inttie genetal fooling inassbull
~
III1thihthe1JyMli~~ Or~selci~5 IiIfieretilltti($trltus teniis ltO -=IiUlteliter~ ~ ~(Flg 8b) ~ lIIithrrolaw~ cCincentreuoos diCIppea framp1~D lBigtif3S
~bUt toaitnpl~lylt4i 150 IIifIo lIIhfjnficw t8st0ted~middot pH
t wi~1rithi thread ClUIps~lMl1BtO 74 (3) IinIt B~ito
785(2) lri-staUc cinfit1at IoihUst ettn88dgtiofthlt ~
it -flnedf81r1y~at aroOOd17S~ lIIlanf1Dlilwu
rellitoteilpft inltleUyJose (by 04 UI1ts at Ii1ta3) but it theo
bull fall only to startrfs1riq tgi1nattM not theIlXper1ilent~middot
bull Dlseusslont f tMresulblmiddot obta1nIid it ~be cl8lirly sean that f(IIJUngI
bull species vethe abUlty to Gtly IIOdlfy thdr loclmiddotmiddot
bull InIl~ts
bullbull L1t1t end dark period tluctuat1ona in algalts IBre obsarved
end ware lri close wpeeIIIen~ with those reported by Terry end
bull ~
Edyv- (1981) with 1ncnasaln both pH end IID2 llIrJngllttt
traquor1ods lIhilst thlshappened ttM surface of the t pre
bull fU_ts ware frelyprojecting into UJe water -deep intottie
mat nearer to the solid _stratlli whl1lre shading fll1lli light andbull accuIIJlatlonmiddot of deconIposJng ~gallIIBtllrial DCCIINed pH valuesbull fell IIIBrkedly Concurrent wlth this dlQp in pH ilaS II fell inbull
bullbull
9
oxygen Such t~ral and $PiItial va11ations in ~andpft
would certainly qive 11se to corrosion cells middotatmiddot the sUbsttatie
surface ~ly resulting in iorroaien pitting
The r] _tabellc ctivlUee or fau)Jng -=raJrwerttmlbullbull
together with their prapeN1ty to-=ci-1tritla~
tlwlsalves wIll also lIied to 1dcIoenv~ lIOdifktIona
Resplretory activities wU1 tend to ~~ tn-the tft and Input caapanants auch a cinia~nY qatk
1mIertabJatesmiddot ara mlk 1bullbullbull middot the bulk of their nlUagllnoua
wastes ~ excrated as ~ 1heexpftunt ncord~ pDz
end pH cningS amN 1nd1v1Q8l~ ~ ~ri
claady 1nd1cat41 the ~ of _tft~t GIl tt ~ of lnIirollliental changes Inmiddot expr_~ta lllhere no Wlter
--nts occurradthmi a np1d dacreUfi1ll pDz
COIJIdalo be a ~decreae in pH (no a) v fladrlg
condlt1C1ns are r-ed tIwn Ieymiddotmiddotor iIay nat be a 1bIIeIqI1int PH
1nctease dBpwIding ~ the --- ofmiddot iheltr ~ by the
organl1n the cluIIp 0wIgn wui a influenced by the
proxJa1ty of 1nhalent or ~a~ the dlnsity IIId the
tIlJcknes$ of the clullp w111alsa t)IMt wiuence in ~
wetr -N_tbull Hencein thick clampllP aUt and detritus
cCUlllulatlon is are COIIIIiIn ~ theh billa IIIdfbls
restricted Br8IJIIltdow1 gf organtc datrital _ter1als CI3IIIIUIIS
oxygen resulting in recLlcing conii1Uons llleasurlllenta haw
shown that sulphids qulckly build up In such deposits and
sulphate recLlcing bacteria y tie-~ SRBconcentrtlonmiddotin
J -I
II II IJI Ii ill
II
-bull I
11
10
the depOSlts woUld terid to Ile lOW at We start of ~r1ments as
condit1onswere nat favourable (POstgate 1979) The cantlooQUs
prockIctiPn of r1th f~plieUdofeacal_te1alor IlUlpllonill1l
~ts bYpoundilt8~ (~~Iick1ngand 1IkIod1) hOweva ~ t H 1 (
IIIOUld rep1dly altAIr theli11l~t and the rua~ofSRB re
notedtofncreasemiddotttcllll ( 1 SRaIll1taat llaUt 1 x 101 SMIIl-1
aftet bull -th
IUctotJlIIl corrosion canfe=t _ta1sln a1Ullberof Ways
(Iverson1972 fUlllIt end King 1975f 1Uilr 1981) pH can
aff~ the ectivity of the 1IIin organlasinuolved the H 1IIho
hava tin qlu pH range beban 10 12 (Graff 1981)
although tIdS -can be lnf1u8nCed by ather fllltors In atI1tl11n
they lIKIJlre a catbon end nitrogen aurCa 11 various
salts tin alactron accaptor (HedJart ~ idbullbull 1985)bullAil of
U- facton are availabll in the debris -alltting b41tween
ar1na foullng -d the 1lUbstra~ 110 providing bull potantlally
corrosiveanvirorwent
Evan IIIhete such accullIUlatiori does not take place varlable
changas inoxYQ8ll and pH ~trations acrossthe suface of
teel soch as C3I becaustd by foul~ ~ C3I give ti_
to corms1an calla lalildlng to piftlng corrosion
There is an initial tllld8llCY for the pH to rise as a relUlt of
the excretoryactivi1i~ of fouling aniIIal and this 1amp UIIllkely
to give rise to corros1an problems Eventually ~er feacal
h
1
~I
materials and detrltus~ill build up around or~anisms ~re bo~
low pH and p02 devul9ls providing a potentially coJJOampiIu
envinx-nt flaw does dify thh _build UfJ but inmiddotthbull
Ixparlunts carrllldotttid notprlvl(lt it R~~r it
l8Iilated the depth of deposit ~1~~ in t- ~ It ahoutd alao r~~- in f~ population so-1QIJvlWela 111111 be dying or dUd Thlb
~lng~wUl~1Ielacal1Md conoa1ve ~iccandltiorJs-t SR8a 111111 b4I fCllllfC314lng ~y pOIi1desa hlbitt~ HtInd 1tt~ WIi~ul create 1~1Mct Jrcx-ntal dlffarenca ~ ~th l1kel1haOd or pitting
n
References
Albilnder J~T 1 bL~1953~~(9EthII prockIctiPn of IdcroaltJodIIs UIIId in I~ studies Rev Sc1 lnstra ~ 528-531
_r1can Ptbl1c Health laSoc1ation 1975 5taIIdIrd~for the lIIIIIlystS of _tars end taatms 11lth pounddIt1on 1IPHIi-~ 1015 ~taenthSt NW 1IIa-l~~OC 2IJ]J6 USA
Bau-8laquo=k1ng C~IIIG amp 1IIoOd EJF1955 Biologlcal ~ in theestuarlna envi~t 1 EcolClil) of the ~ cycle Kcnlridlkjl Ned Akad -t ~ Ser B~ fi 160-181
cter DB amp SUver IA 1961 In IRafaranctlElectrodl$l EdltolS Iva DlG and l-z Gl lc8dilll1c Pr London Maw Yodlt Chapter 11 ~
Davies PIII I 8r1ric middotF 1942JII1c1oel4iitrodas for-aln local oxygen tension in n11 tissues Rlv Sci InStruII Yo 524-533
EdyileariRJGJ ~ Tetty LA UJ83 The 1nfluenc ~lIlcro algae on corrosion ofstructural steels used 4l~ the 1tIirth
Sea 81odeterioratian sect 336-347
bull bull
~
bull - -- ~It Jf bull
- ~ 13
12
Terry LA I~Ed)VeangtRfiJ 1981 lIi(toal~IOI1CGrmianF~ l)i 1St1oJhemailnEifooiiiigffixed gftshoreIt Bot 1Iar ~ 117-183 lsect~~jlaUoQ1l1 _ ~t Ef)e~gy Offs~r1I Te~lOtjjy Paj)8r
-middotltcntpI)~it$t~l91-16PP~middotmiddotdl ( I
bullbull
iIIalk e1 JL 1971 bull Ion specfie Uquid I~~ e)(charig~ ndcroelactroclels Anal ChIIaI ~8S-Q2 AGraf1~ w~S 9S1~ 1orrosloh InFIntroooctI00 tooffshOle
51~weSoes~~fab~lcat~onIns~allat1on Gulf Wall PI aelUnger EG 1982 The appl1~tlon 9fMl1ikIngmiddotHoust(jfW~Q23g-258middot icroelectrade in the study of pH end PD2 in lIHinli
HerbierC SN fGH~rt~ )RI -Stoctcdalpii Watltif1Sal RS ~1ronIIInb Ub~~ 27l-Z79 1985 Fac~or lontro1l11)9 tha activity ofmiddot sulpMte
bull Wall P 1983 The appllcatlon Md ~ ofiwrJ)e~c~odl shy-taicfii9 tiiCtetili tn~ diiilncj tar infection
techniques for the iiaurnt cif icraeov1ror1liltital5QiCiety ofJebo1- Engi~ SPE 139781-10 _i~~ - ~ ~ i ( _ IKIdif1catlons caued by aGII8 rlnefou~lng -C~ebullbull
PhD lhaaia Ibiverslty or ~tar UKbull bullbull bull -
IIH1nke JA 19fiSa In EXperllRants in Physiology and middotampioClIiIiiilatJift l c E(If(orltGAiKetkutgt ~c-Presa-
Wooltdngton AObullbull Davenport J 1983 pH lind IlIl2 IMla ffl~- j~t~5 -lMIIleath 1IH1na ro-fauUngGqWl J bull Expbull 1
B101 Earlsectsect 113-124 Klnke JAIII 1969b II) Glass IUcroelectJQdes poundx1tors Uwal1~Ui~ OrOllt arid~t Nt JOhn fII11ey YOI1lt thaptar 11
~-f _f~ ~ ~~nt ~ Igt
bull Iverson IIIP 1972 Biolog~ Qrroaian In Advencesin
corrosion scianca and tecIYIology 2EdS FCntaia G~ I bull IIS~et RIIIbull p~ PJaSsLancIoIl 1-42
bull
Jorgensen BB Reusbech NP I Cohen T 19B3 Photosynthesis and strUcture of bIIn~ II1crobWp MC1OIIlectrocki and ~ stud1ea of four cyanabacledalt -~usllWtOl ~bullbull 1~f093
t l~ O~ ~~~ H~ ~9s7~c-gtHatste~iecfn1k van shy pellediftlrtenPt - Stlchelektraden (1-Su
AiJBitiKlificfiliilislflilr)zlIr polrapusdiailJIIissui1g_ liIserstoff(jruQkas fur d1eB lIsUbullbullung der IIliJlttOUf1cillatlon PflftgaHArdI ~fl39
bullbull Mller JOA 1981 tals In EconcinI1c II1croblolooy sect McNbUdbicldatariOtationEd Rosebull A middotAcadelc Praulondcn149-202
t Mller J DAI King RA 1975 8io$~iorat1on of lllltals In lIIi~1al ClflPlCts of the deterloration of tarWbullbull EdsGilbertRJ~ middotloveloCk 0111 bullAcadeic- Presst IILandon 8Jo103
bullt
IPotgate JR 1976 The sulphate-reducingbacteda
Callbridge tIn1versity Press Cliilbrldg8U-K
Reusbecb fUI i Ward DIII 1964 i~~~1Ida5tud1el of
bullt inter$Utilll IIIItez dttm1stry and photosynthetk activity
in ilflaI spring Idcrob1altAppl~EIi~ MerobW tsect4IlU-27lt I
IIbullt bullt
bullbull Ibullt
1
Inmiddot middotmiddot11middotmiddot f~~ a~ -tmiddot bull J~f
I~ W
i5iImiddot J~a t
ilmiddotmiddotmiddot I middotIt Ii
l
~
middotmiddot~~middotlL= g r
Lj Ic
t
i ~
I
gi II cEi ~ I I
1 t rW
D
tR bull
bull t 10
11
r
i
I ~
j
~
I
~bullbullbullbullbullbullbullbullbullbullbull
LV
FigS pH and p02 profiles into macrofoltling eomlllllllitilS dominated by Balm trenatU$ on steel plate 11 two pH profiles Ib) two lIOai profiles leI pH and P02 profiles into dead barllllCle shiella withdetri~Us I I
itIbl r p02 ~ I
nvnHg 11 70
j ifJi eo 1amp0
laquof it 30 s
I 20 N
ca 11 10 a o -- shy i bull
11
80
19
781 o 2 6 8 10 12 0 2 6 amiddot 10 12 dIpdI Qf ~iftto fouling In HighIst numbIII pIatIi
8deg~~
~i c5 iioo5 fiil
pH 8 bull_ ~
~~ ~
_ bullbullbull- pH
-pOz
(cJ
e-Clf ( j~JJlaquo1- --~I ~a ilil - _ I i ]1
~ ri en i ~ oJt ~iJ id tUI
2u1U I shyI t
shy1 2 3 6 6 7 8 9 10 11 12 i shyDepth 9f electrode (mm) into fouling
1 i
1
~----------~-------
~
~-~ lt1~
middotmiddotPmiddot ~
I
V
~
ill
1 Tbellitt~~ of cathodiC protection OO~~lIic~tlO aridrfOflllUlc
ofi AA~Uoullr14l ~1~I
I
ael~lner EG middotIUtcb8l1AJ Dept lt4 lInY1roMe1lal Bl01011
=~~venilt7 IlariCheW 1f139PL $ ~11al1df
r
I
Ablitrict ~~~ e~l billttI ~ted with ~pert1n ant11_1111ampp paint llere itpoii~1n _vatI- In the tl~ld iUIit 1bOioatbrt~ScM Were
1IUbjll~ to qtbocJ~C ~teltt1on byen _ current (~)SJi inci EDAX Iieamp~ta IJdlctecI ~ lachln teoc copper OIl tbePlCtec~ pa1n~ atter14 dQ1a~ protected Pllnt abo ebowed
) reduCed bacteiial activIty butmiddot ~e4 dltoe Iu1d protOzoanmiddot co~~l~tlont~1l IlU l~byenCOCltact not byen dUfwIl00 ~r _baRl lrillolvedln tOXln nil ereio noted
l ~~
1
I
Introduction
Applications of protective coatings and cathodic protect1on(cp~l
are recognised corrosion control methods for IIIBrine steel Itructureso Cathod~c protectlOll preventl conCIsion by Int~lng til8ctrl~l
currentS trem externalllClUfCa to e~~tthlJno~ e~ carrollon feetlans C04U lUChu tUauJ1ng Aelnts a
(
barrIer to ~ the flao of couos1a1 ~aboIHnu~r
Dccurrlng anodes cathodes or wlwit gelvWc CCIIII1 ( 19681
flinger 1980) AnUfauling ~ also JJICOqIOrII~ b1oc~~
Tr~ylt1n nuor~ (~) end o~ Oxide (~) (~ flJ1J
- of u blacld1111~
V _Ut Evans 1981) ~ ~
end growth of bipfoullng 0I1I)IIIII1IIIIe Wdd~ ca ~ of
paint rusendmiddotmanemiddot ~(r~ til )
As corrosian ~t1on ~ bICCIIIII -an ~1tlQd ~ of
antiroullnQs 1 conjunction _ith CP have bllClIOr ~
resultIng in reseudt 1rwestllj111tlng trw COIIIPIitlbUlty or VU~ ~t t _~
rOratlation to dlfrert levI ot CP (~Eelf 18110 ~
~t 1980) RelatIvely 1nIestiljlllUans have ~ urtw ~
or CP an thIt anUraullngpert~ of palnta
In thIt preseot 1nlIestioaUon tV~iln111 ronutlon ud
cOIIIHrclallY incanjlntlon wIth CP was ~t4IC~ Iw I sraed current (-966 l1li) arid exposed at the 1 Strait IInQlbullbullbully
~t1ons weremiddot 00 the IIhcIrt bnifects the PprIUd_
had an a) the various COIIIPQI1eIIts of n pe1ntf1l11 end b) l1oft~
developaentbull
bull bull ~
~ ~ - f
bull ~ _T~~-middot~ ~~- ~r _ -$1
(1) ~rlttionor Painted Steel Billets middot_ ~~tmiddot_tlJ middot~hmiddot~--~fmiddot ~ middot~A~-~~middot - -~ ~
Thirty-al ateel blllets (8-10aD2 end l depth) were cut frollbullt ~middot~middot~middotmiddot~middottrSmiddott ~jS-~ It bull gt~ tfl - middot--imiddotmiddot ~ t1dItrd steal plates The dimenSlonsof -thB butits-lim chosento
bull -~~ middotmiddotgt2h ~ ~-Imiddot ~~rt ~- -middoti~~ ni ~~middot1 l middot-~t~~middot _
(a) fecilltate amiddot Ie detrucUve -thxI of prePraUonpdor to ~~_gt gt~ k7i ~_ t~ 1--1- _ ) - +--_~ -_~bull ming electron ~ 4II1CI (iialloWdirectobs8iVatlCnS of the
- i lt-- ~ gtbull -o1e billet 8Uface
gt r~middotmiddotj ~L(~middot~middot rmiddot tmiddot J ~ - ~ - lt _ bull Single non-standard (1hIula~) nickel lIIireSmiddot Spot Welded e1Cing
bull ~1deJ ~ b~i~ ~j~~ tri afl~1b1ipOi~ f ~1t1 ~4middotmiddot f-gt -~ ~S-- ( -- - - _bull resin EactI billet then Undb~ tI1d prtiladlllltha5O __ eyilr
bull ~~_ ~~imiddot ~1iil h~ intlfcili~ ~eonbt1nlyi Tr1butylt1n F~id(18n)~~~O~dettu20i~blcicld8is 1IIfii bull i - ~ ( then 1IPI)l1ed to glvaadry Ulil tIi1c1ltnOss or 1(1)-150bullbull
tbull (2) IipreSSeCi Current Syat
bull A rigid ~tic due 1I8S drWedllliihwrtv-s41 liiIihOieS IIPProX~
bullbull 05-10 cia episrt ~ Pilnted ampUl8t Wsfbed IntaPQSIUon ampy
threadlng the nickel IIIlr8throu11 the dinto a tarproot junction
box Eliltilen of th8~1l1ii (recelving ~t-d curnritlere IIIlred
Into bull block connectOrrecelvlng dc trOll is potentiOstat The
rainJng iontroi blUets Wet8fbedfl11i1iiy lIIithln thejtlytlonbox
i~ityd~c IkiliPly ~~ ~lat1ruaWiode 8ncIacalOliel
refer~ electtodetoiere fllied ~ the per1aetat of the plut1cdlc IIIId
connected to the pot8ritlostat The desl~ of the potentlosbt ailoiolEld
~PlDteItlO1TClJrnnt to be MIllntained at 8prampset levtl of 966 II1II
c
r~
Grid raf 1J5s-t 114iH 563724 dud 0 Juna
Alolecule andbechr1a in general are ~ to
sutface lI1eroleyer (nlUSton) of _ter (~bullbull 191KJ)~To avold
contaJnatlonof ttw painted surf by U- -vents
blllets Were s-nact in a 1 conte1ner or atc1le (02i ) f~~nd
ter Iih1ch then f9ICI in tv 1IIhere the b~
be r-oved undeNeter El~~ bllleta fa Mcb trtai bullbullel
removed over a 1 dtq (336 hr~ expoeure perioda
each being ~~bn uposurnof 1 5 11 iA 168and DlIIauz1r
Arter field axpauHli the billets wan
fUtered _tar to ~ 1_ debtlflMd far 10 aJnuta ~
wv ~~~endattid sdbUiUd(otO
~ ~l f~ nn1ng el~~ --
lOOted onto 1middotCIi ~bullbullWIsend carbonmiddotcoa~nmiddotmiddot~middot f
each billet Ws~ Iis1ng a ccr1dge ~el~~
(5360) IIIlth anergy dJ8perslve X-ray alcroenelre1a (mx)
_lysis the les Wen reNed end coated III1th vold to al]Ql En
detailed sat of the alaquofa1l
(4) Lfiborattux ~
Painted taa~ bUletalllllre pnparad ~
in a Mrlne reclrculet1ng flOlll _ ~
f1Oll the ~ ~a1t~y Tha~~Un bullybullbullbull
10C M1nta1ned ~~t1as of ~t~la protazOllbull~bull10shy
(Entermap) ~ ~~~middotNlt~Eamp1~ __~and ~ r - ~ -
certaJn ~lc 1IQ1~ 1(1 the
the painted
~
thre repl1catft fn
plKed 1nto(022_)
- thIine1J~~ -lt bull
Atter mAX
(1-2) and i-n-t bn collected
14OC
Pain~
-
12 daY~ exposure and prepared as in d)~ - t
~I ~~~-- i~yen 1~ v
Rflltsi ~ _ ~f-~lI ft j bull _
rt~ll0~~~llY~~~~peln~stetsl~11i~ex~sed for
y~t1~J~ ~~~~1~1~~1UII~ya~kooJ~~~lt ~ ~~_~~I1J~ ~~~tomAX -lysis ~era than gold ~ bull bullbullbull c - _ bull lt
~~fllr ~~~ vllUll ~t1ans of the rfee htures and gt Co
~f~ - ~~~ ~f ~ ~aobta1Mdue IjIll811 iI1Flg 1~2 The IIIIdn ~~ ~ lt)ji9 )tmiddotmiddot - ht
- bull
~I rIIplt~ tIt8 -~ cOlllponanta of the pei1t 1bullbull WelIIlItr1x $ilj~t-~~middot - ~ _ -_
-IUch PVCbQed end rap~13y the chlorlna peak the
i~~~-~~-~V~~~-t18 ~~~~ ~~ Th8
ral~~i~ JlrQP(lrUo1sof ch 1Il-ntal paek~ll1d l~t_ Q~7 -gt- ~ _~ -- gt ) gt -_ -lt ~~ the 0 -end _ xprtsntabullTIwsa changes are -r1sed ~t~ lt~ _ ~ 1nT~ IThe8llia$lb npresentthe chengelI 1n~t1V8 p~lons
ti~~~ [ ~gt 3 - - - _ - _ -
Qf nellllllllt within We surface layars of the pe1ntmiddot ru The
~f~~thtJn -d ~rfeli avr the 336 h11cxI ampScanbe
SII4iIl r~ fig 3 al~ tht rta of~ dirfers tJetwaeo thaa ltmiddot~1- 1ft~- ~ _
IJOl appfiai-ed to ~1nlt1elly until ftar day 168 it fall
~~l~ 1h8relllllrelgn1fi~ diff~be~ qrOtactlid and
proieCted (at -966 ) piIIllnts tor lioo COPPer but not folimiddottin
Ibi~tlons Of the Pain -urface 8hoWad _Pil ~ witilt1M
(Plete I to IV)~ 1hePllat1na (~) pelnt(Phta x and b)
Clearl~st-i~~r alots on the SurfllC8 IiII1kh~ter1sed ~- f hlt1 lriitUl tin conCantrat1on~ lllariy slotsre Partially
~OIIe~ ~ tile _tr~tedal middot1lere rs ~an~ densely packed
grerltr~ ~th 1ar~(iIa1niy cotifIIr) 8ntt -ampniBil (lIB1rily 1rori)being
middot vl~l~~~~~ Dtt~i~~~t~ r~ ~f~~ I~t seawatllr for 11 n IIIOre slots appeared and th~ ~~ess _trb cwer
--if -- V~ c~~ f(i ~~~i~
(Plate Ie to It)~ the granules dld not protrude from the~rtace in ~ ~~~~j1~r~-ffJmiddot0~~
suhap~~r Both 0 nd --960 at ~~pa1nts ~ - fr ~~~~-w1-U~ ~~t
sl~llrat thi stag~ (Plates Ic-d II1d xrnspac~IlYh middotmiddotWith middot middotaJ~cmiddot gt incraallO ~alolre t~ difrrqcbullbull middotbatIiI8IImiddot the protected fjd
bull ~ ~ ~t~
IIllrotacbldpaint ~~lIfIt~ftitr~ h alQt~ p~t bull u bullbullbull J ~~i~~ 1~~
lIIith IIIuper lIdguon u 4lrotatad piIJnt (Pl8ti1 II) 1hD1II8S
Ill )1IMdw1lillS gnnJla~on f-r~~ dpert1cl ~ - - - ~~r ~t
Onthe 966 1IIpetnt t-var tIIO typeaofan with diffannt c t
~hIId~(Plate UI a to c) One hIId tIII-1 bull bullbull e _ bull
fUll wJ1hleas abvlaus alats the o1fwl bed C18UU1IlOt ~litUi middotC c ~ ~
less ~ t11 h Gnnulatlan __ but ~lIiIZly def1nlld in
middot~ latter Afte fila bed tiIIrtIId 10 ~are ~~j(
extansll(tlly ~ the~tIId(O at) tIIIIple lIIIdngthe~ - ~
middot orarallet1ai leas claaI (Plate I~) f u II1ao CQ1tJAJ11d to ~Gn - -
the -966 at 1 (Plata III d tor but 1JI8V8n 1nittNdcneabull -
Althatqa -middotslots wrs abvlaus becaI1ng 1 cwampr aiI- the orwWat1an8
f1nailyftar 14 day exposure ~ fUll drMIlapIIIent on all la - ~
are extensive with the 0 lhawlng both alota pore ~
OVIKgrOloln (Plata II c to f) Gnrulatlon 1bl to ~ gt~t~~ i
or the th1cknats of tho ma ~the ~ at ~ (~~~~ themiddot - - _ middott ~ ~ ~ - -1 1 t
middot f11bull IIIIIS IIOre extans1 thIIIn at dIlya but IIIIIIIY bull1Q~ tI st11l ~w -middotr~ 1gt ~1 ~
obvious fIIany ~ loI8re still p~ yen~~~~~~~r
than before Poiu appearedto be f~lf~ila~~c-~~~a (Plate IVf~ label 1) IIIIl1ch broke~i(2)~ ~~~~
t t
bull ~lS (3) ~ rflC~~ ~i~t~~l~~KI~ t~ldiIiiicyul~loIiih ~~Rht ~ )~- -~ - ~middotd 40 ( gt ~gt ~d lt1 t ~ Jt ~remiddot4amp tha paint trix cavity bottom eroded away reVe81~ng ore
t ~~t~ por~~ ~l~ P~]Y p~~8dby di~lutiaA~ ~~~~ti~Jr~gt~ta(l~~Siinll1cl1Ud~f iUiIiI reriatlGllbullt
bull a~~ ba~tid colCriAtloriccic~ GIl ~~~d ri -966 - ib~~~amp catQn1iti~~t~I~~(~t an ( ~
t the ~~~palnts (PIa_taiiI d~~h ~wh ~ n td have ~ CamplluJMitiIVc) ~~middottti o~lgtnalrf tlt of hIght
~ l=~A ~J~rW tYtles~ p~fiiilriQcOCCold to~Ja to jU-t ~jmiddoti~ai dlatClilS ~ir -jlati ~tbullbullent aNit ~ t ~~ th8middotf~ld~ies~~t~on~ ~ -966 prQ~~~(P14ta Iii~)Omiddotiliof~m1hiiHAiiipieat ere~ll-(piitir~ p-iot~iilans(PiA-iII~ end c) nbullbulltiledt dl~by1i~~ end wirinali1sheii ~cMI~bY thI tili
bull In t-~~ ~1i1nt 1oIIIeHfd~ ~m1IS (15OClind a t ri~r-dlatoa ~lor P~t dlataiOlonlt1anGIl ttw-966
t
bull ~
pr~t~ ~is~ih iion obitJauIJ A ~rot -=1 ~ ~ i __
reprasenttild on the 966 painted 1iUrfc1ncluicunljj ~ rwt
Achnanths Colonisation by bacteria 1n the laboretory bsed
bull eliperlMnt tollMct sbiUar pattern to the fild ~
bull bull Dilicueii
bull bull DUt 1Iidnt ~lona Mve dlttllrint ~~ ictian
It ~ JntM l1tenture ttt Iff yen __ JIll the
bull ~ bicins ani 111 ~t INchlnil (PhillIPs 1t73t EWene~ 1981
In ci1ntlHt tJWY ~tit ~Wl1c iIuCh as 1rbiltyl tirI
~ lite wAlII]ltiClr dlsP8tfu~ middotthiirsttilll~intaJn~ a
bull ~
lIIifoflll ampalCentraUon through the filII 10 thoIt biocide rel_ at tha
paint surfce 1fI$Jlts in IIIOre ditfusing ~s to repla~lj -iflJ~it
resulting 111 bullbulltNdy lr leechlng rata M1llat this IIIIY be middottruIi ot
tin ~ it was nOt true ~ ttw middotlBlf fOZWJlatiGn ua ~ bull - i - bull 1(1
ObSfvt1onundr SE clarly ~ tM t1l1 to be present
~ly lIS OIIIdle-llk parUc1~~t tn lUlfiice (Ito
IV) bUt usa ~ the dIpth attt fU-middot ~y~
If one considra the concn~rtlonmiddot chMge 111 1n ~111c COKICIl8rlts fro- the paint tU- i bullbullbull tMWei b~ ~ Si6t arid
tI-~t iron oxidt 1~ can bt (Fl_ 3 IrId TIibl8I) ~t ~ ase cona1derllble d1fftrenC8s ~ a Un decz bcdYMr ttw p1iod IIIIwraS lron~- an unt Ught inCr h ~ incqJper ~tlGn occUrnd _idly at ttw ~~ Gfb ~ period ~ out ttwn fu nPldly ampQII1n ~ umiddotirid II phast This IIIOUld bt tIlq)8Ctad It cantact l-=h1ng we occun1ng
IIltfIOS8d pertlcl of copper would lMch rapldly on t-n1on 1n - shy JIsewtr There would thlll1 ba bull J-a a1iltUy leu avalUba
i1 ~ ~
_tedal becfM uposed Thi tIOUld ttwn IIllCh Thla pulSing efttct
IIIOUld be expctad to -oth OIItlllith U- putic1 __ -lllible II tot 1~ GIl a on r-x _lao Thla teet 01 put1c1e 1tUng
can tit GIl the lIUrlace ClIlC8 it ~ a pan 1amp t
a 11aM bet in Plat I to IV IIaIrIg _ lit la_ II ~ 0- ~s _- PIate IW $llldl ~bull nshyot poRI flllNltttn Al~ tin ~ lou rete thtn I bullbullbull atill Ivwnce of bull al~t pulsed 1Itue rrlict tII1IIWd
elqlCtaG inIl ttwauUr ~ I I I
0
~ i
~ ~
~I J
bullbullt bullbullbullbullt bull bull ~
bull bull
I
bull t
~~
The same seqiHllle of cevity follliltloii (intttecaseofthetestjjltle
slot- shaped) fClliOileciby SecOlld8rYtlolesfotlllingin thll cavity
bottollis releasing further deep8rtox1ils (Plate XV d and g) As
cavities_re ProdICed ~ tiecaaIelnvadedby_ter~ onelilauld
expect 1ncreaa 1n1IOdha and dIladne lMls t the t1met
which the toxlM uw ~ Ths fCMld tCtHi eo (Flvmiddot 3)
~rttno the vi that_tat ~t1on otthe~ ~lY
il1Cl1l8S8S Idth UIiJ
Differences f~in toxin li~~to Ini 966 ill 8811P~ F ~ bOth Un end ~ lcMIr levels afillW
IIIIIlfI ptaInt(~1~~1ntha~~ ~~f~~ cst rklld with tbiIccpper ElJIc~l ta~Uona invalvinv
w pl8cId at a~ht~~middot3lO~lth Un cqJpel bull
(-960)10 ttW ~af this expei1tIIri1ts thIsiorelfJltdt -
that cqlp8r b8IiIId ~ IIiIOUldProc-ln f41ip1dly t thellllgllitiv
patentW _lied () as 1t 11 further f~1t ~ witt Un
III11ch is u-tat aqulllbd Idth
it If ttda 18 eo~ ~
~ )
frOll1 paintscould bl anqnced by an illprd current clithodic
protecUansysteipos1bly ahart_nino te 11fe of the paInts
antifouling ability Bacterial coloni Uon IIIU rtdUCed on the -s66 paint This
could have been due to unflSllOUlllble electrostatic cqrge on th surtece Th1a effect CIIl pley en IIIportant role in the t~of
bacteria to alrraces (lIIarsMll19E1O) As bacteria appear to IhiJw
greater toxicltyto copper (DMpsey 1981) it is Klre likely that
enhanced cqJper leacll1nl On the -966 AIIIPl c8used the recluCed
activity The IDlol8r toxicity ofUn to bacteria (as reported by
~ -
DelIIPsey Hle1gt or its lowes leaching rat was sUpPorted bythl
relatively rapid recolonisation of the slots by lIIicroorganiSlllS ~ftar thE inlt1al aurge of leachi~ Thailllpressecl current dto ~~ the oppoSite _ff~middot ~r on dtta colonstJon l1lI-1- in the
JIbcJrtory~ta larger rUDIIra f~ fit pro~td ~ther than uriprotected Pafnta
Ccnlus1one
1 tiild-a 18 presaIIUd ~ thattJn~ ~~
~llNChng notd1rf~ ~
2 TIleuseofCP~ ~CUIftnt(at) on1h1a ~fca41ng ~- ~
pa1I]t~ to ~_lAIIach1ngn_ at toxiN - -
3 leIJstaPidbectar1al ca1on1aatJon acaampmId an ~bId Pt1ntJ ~
4 JItore larger alcrofauJing (Oiat_ and PzOtoaoana) Gr~_
occurred on the protcbld~
I l
I
bull bull ~
mfEflpoundaS middot~-t9i _jgt- ~ _ _ ~sev M~J~ (1981) (cilonJzatiooof alUf~lng lnts by _rine
Jbull ~teda9g~1ta IIarina XXlV~ 185-191
- - gt 0 Ie 1 _ - _
poundV8I$i I~lf (1~P~h 1II8rin~Ugae and Foul lng AReviw ith
t p~l-i9Jrlr ~iiMdilto5hiP-fQUllfCiH 8ot~ ~r~ ~~~ XXIVbull lIPwl~J11A
Fotteth CN~R ~kk8nGB IWPIR1seJCiiiroaic1lihd ~e ~JIIth CII Offshore St1ucturu ElllS ~tjI~f~1Dt19i4
- - ~
UKC (1976) I~races ill ~W Ecology KIuverd Univbullbull middotraabullf~iddge ~bull C~~jII illt)Ia(t RC (1980) totinllsa Cattod1CPrOtect1cinbull middotmiddot8AtEmiddot~6haOCIlfif~ T~ p No 108 ~ 18 ~ 1IIHf bull
t bull lt ~J~~~ __ ~
Norkr4irulB(198D) Advanculn UcrOfj~lpound~ltiOyVol 4 (AlItxlaquoIder fl Ed) PlerIw Press lcxIdon~ lIP~ 51-85 ~ l~l - bull bull
PJijllt~i At(fm)~mr~~JnI~1fqul1nQ JlalntlJ Riumh ~JnOrpnic toatJngs 2 192middot
)~J gt0 c ~lt -- ~ __ RogerS~ Ut (1968) ina ~(lnt~fbItliP4ili GI COnoslon ~a Technology)bull bull
Sl~iorr~ VP~Ranald~ Rctd~~ (1S80)_ ~ Experllattal Stud1aiRelate middotEffect or Cathod1c Protict1Oi withmiddot terta1nr1c
Coating Syst~ 12th Annualofr T8Id1na1ogy Co1fItHnC8 liauston TIIlt p 9-12
j
fIG1 sPct FromMon IT8TJCuZOI Int Lliu EOAX
FlG~ Fro TlTCu20Ar-4 Dy Field Ellposult I~ir
il
I c
R hi -00amp=7
-I I I I I I I I I f
iii
bull bull
bullbullbull
I J
J
I Figur 3 Thl chan In reltl rqpoIQnlof -ariOtimiddotjOmponents of the paint film with time Thl vtic~1 ampkiS unl till _n ofmiddotmt Illment per unit OJ fe 50all Cl Na
Cu StI Feeshy
PlistinllFe S7~J6 704 3~74 908
f
r e ~ ee~
middotie ~lt - ~ shybullt bull ~
t -~ l Cu~ o ~ f
t --- 3igt - bull 0 J Sn
cc
t ~
~
bullo
~ bull e
bullbullbullbull bull ~
bull ebull a QI 1- bull 0 40 431282 316
101 dmllIII)bullt bullbullbullt
hr 0 ~~bulla 5~16 ~~ -~~ i166 4S i83 8~1B
24tu 6~480 SD~83
111middot 1~1 OS6
~ 5114 6~69 -51 UIamp middot016- lt ~rmiddot
1d o Mea 818 21 021
~Z21 199 1~1Qmiddot ~Jgt2 058
14 d o ~~ 804 Q~B8~~~a~~t~middot 685
866 372amp 8amp4 OS _i~11[ ~~64
Tttile 1 A81UvlL pzQJJortlcw 0 vxl~oI ~~bull Jn 1I1Dtitctel lt-1166) end lqIlDteCt6 (O)middot~af~_~middot_lodEDf~ middotmiddotmiddotmiddotmiddotmiddotmiddotmiddotmiddotc
~ ~
_----I~
J
I bull
I
J bull tmiddot
7o CONQRESOIMTERNACIONAL
I D~CORRO$IOr(IAAINAEmiddot~USTACIONES~v c)
t 7thflfERHATlONAL CONGRESS
ON IARINE CORROSION AND fO(JUNG
~CONGRE$INtERNATIONAL DE~CORR~ON ~E ET QES~SAUSSURE$
i t
~ ~
bullt
t t
UNIVERSIDAD PourtcNlCAVollncio7J1 NiwiemIif~ 1988
(SPAN
SECcl6lfn
BiOIogro marino Morine biologY ~1i1orine
Theunpub1ishedJllBIlu8criptEl have been reprod~d asr~ielved from the
authors The SCientiUc COlIIIBittee take not respoi~ulbll~~Y for any shy
error or omssi9n
Laa ponenciasno han side publicadas y estanreproducidas tal y como
sellan recibido demiddot BUSmiddot autorea pound1 Conte Cientfico middotno se responsemiddot
bil1za de cualquier error ~ om1si~n
bullbull _ bullbull_ bullbull ___ bull $_ _ L _ - _ --- -__
SESSION II Marine biology
Friday 11th November
SURFACE PROPERTIES ON NONmiddotTOXIC ANTIFOULING PAINT FILM
K Nanishi M Murase Y Yonehara M Kishihara and T Hirama
MICROFOULING ON ALUMINIUM PANELS PLACED IN THE TROPICAL WATERS
N B Bhosle K Venkat and A B Wagh
EFFECT OF TRIBUTYLTIN OXIDE (TBTO) ON THE OVARIAN MATURATION OF THE PRAWN CARIDINA WEBERI
R Nagabhushanam B Indira R Sarojini and Mary Frances Thompson
EFFECT OF BIOACTIVE MATERIALS FROM SOFT CORALS ON BACTERIA ASSOCIATED WITH COMMON FOULER
Vitalina Mary Avelin Mary R Sarojini and R Nagabhushanam
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
I bullbullbull shy
-
if Ilt
JII l3$i]J ~e-l~ ~ 1
J ill KItt
Ii I 1 8 Ij j iJ)Sj
emiddot Sllll0 r B 8Je Do Ji
t ~l u Ji ]
1 0
j
j
r 1l3iiiJail1h
bull III
~ i J ii i )1 t~I ~
~
~ )38 ttl i UN ~I3 11 I ~iii bullbull I fi~fi IelI M 6 6 bull s plj i I Jjhi
~l -u II11 i~ Ji ~aJ j~ ~ tll
- bull
-------------- -- - --- -- - -- shy
2
This paper desr1blS the use of ti1c~lctrooes to measure pHbull bull _ bullbull ~- l bull bull
iJQ2 chmgtS ~~ 1II1C~olones ~middotilIlg~ ~f~~s i~rsed ll ~ter ~ wlJlch t1)U11ncjotgaI11sllls are grolog SOllie p~tnt1al 11P1~~tions C1f~se cIIatlges to corrosloO at the
lt I )
surlP alllO br1lJfl)dl~ - 1 - ( _ ~ -
ItIiterWs 8nd ~ ~ ~
() poundI~~lt - -
Ttl lec~_ 1n theu bpelsnts hIM bnndeScdbed In
~~~I )iel~~bull~~i~) but n 11 ~ haJe
- Aj ~-
(1) pHel~
ThapHalec~ lII8rII basadQIJ tne-iWof It1tQ (1__)
wre the pHlt1V8recenton of~~l~ 1a fiI)OS8d bull 1Ihart po1nt extlndlngfro- tt lnsulatlnQ (Flljj1) The
pH andmiddot alUlllno~nlcate glau 1cropipettesued lnthe - - ~
electrode constnction went ~ using bull wrt1cal alectrode
puller assntielly bull vart~c81 version of the equiplllltnt
orlQlnally developed by Al8Xllllder and NasWlaquo1953) The ~ ~ -
electrolyte u-d1llllS 1N tel and filllnQ IIIIIS 8aoIpllshad using bull ~ - or
aydnge 8nd a vary fine draIIIn out Qlass capillary
Once f1lled the elfCttocJes were soaked for severalday~ in 1111
tel to allow coqlletehYdroi~s ofthe pH glass lin lnternd refelflf1(e of sllver lrecoated wi til sliver chlorld~~~s
ccinmctedmiddot to a gold plated pin iealed into the imd of the
electrode lII1th~ill~oo rubber COMPleted ei~trodei er~ stated Up downIIIards lna dilute solution of ~r~iIcrcj In allmiddot uper~ middot-temal reference Qf bull~~l
allCtradilllllS UIIIId DurlnQ -~ts -n bulllact~damiddot was c~cted to ind~ ~~F~Y(_ itrgtbull The cStcu1t layautforbotitthiiptl lIlIdpDz electtaifes
~
1a IIhIWi Sn Figure 2 bull
~ -~ - ~ -- - - - I -
CaltbnUariof all alacbodn id Clirr1ed middotOutmiddotbllfGn ftc AIIICPUIIIri inmiddot c- elctmdeiimiddotmiddotmiddot_middot~
ckalng en apars-nt by in nJIalt~ orthey dldriat
pEOCia ~lbblIt ~lntal callbraUon In ~
CIIIIi8S all ~ datafl(WtMI middotmiddotdl
(11) 0ItygIn a1acUocfes bull
TIUI catheter typa elachode use a fiat ended and of duw 1 to 15 _ It conslatad ofa ~ uia P~tJft- wlra
aeelad 1n 9l~ and IIIDadded in 1IPaKY~1o 1I1th a aUler wue KUng a raferance (see FiQ3) Ihen the epOxy hId~
h I of h l~middot careMJy ~ flat to UpQM
bath the plau _the ~lwr 1IIIIIbnre WIis ttn8llPlled
ecrosa ~ flat lind by rep8atedly d1fIPlng Jt into a aQ1utJDn Of
collodion 10 atheta1cohol A Pola1-1slng voltage of m all __
applied bit eachelectrodl and these re not thariuSed for
axper1lllental purposes until middotthe response I118sstllblallihen
bullp
4
bullit tuffer elut1on a~ a eorustllllt
it i)
~~ ~ ~eted IItld accepted for uSe elliCtlodes
it lt gt ~ ~~J ~J _ bull~ - - -gt bull
IIIIiIr~~Jn~IIOWt1on of 02 KCl D~ygen Ilectrodeser _iliry~ ~~)~ 5middot -1 ~ J~- - f -
it cli~rated aga1pst solution I of known oxygen content (as 1 w f - w ~-l _ bull bull
i IllllllaUred bY n of the l111nkler titration thad APHA 1915) ~Lmiddotmiddotlmiddot ~ ~
it Any el-=trodI not lIlCtIib1ting a ilnaar response rejected ~ ~ ~ ( _ - ~
zelia axyg8n lution obteined by the usa of a buffer
i aturated wUhpyrogallol crubbed nitrogen The 100$
~~~ reed1ng obtained by bubbling a1r thlOUltl the ~ ~~bull ~ ~ bullIII burfal far 12 hours at ~~ture 8M pressure- ~t bull
(b) [)cpars-ttal~
All -rs-tts IIIIIl1I Wried aut in atellperature controlled
bulli it
tank pliad an a teflain NYler ted l~ steel patfon ill insidil a ane _tre CIbI Faraday CIlIIIi(fig 4) Tbe~
prav1decI IIf1th a lJrfQII hinged door at the fiant for eccns and[f as gro~nded to earth to scrbullbullnout radio frequencyi intelfuwce The _tar flowing ttmluitI the experluntali - tri pullPed at a controlled rate fro a temperature
~ controlled reservoir For all experillanta involving algae
i llllA1nat1on provided using daylight fluorescent twes if l1d teel and 1Ial1ns plywood plate approxilaately 20 bull x 15
wire Ixposed in the -rine environmantfar several IIIQIIths to bull
allow a heal) growth of IIIICrofoul1ng organi_ to deVelop The
bullI steel plates had been placed utldally and IIIItre exclu8ively
bullt
~~==1$ and ~klOOrganl_ ~ldirig Itltena ~srar-nt of their
OWl The wooden plates 1IIIIIl8~sn the Jntett1ifalteclJQii
-f llllllre COIIered lIIIith ta 01 tt- alp InWfCld1IIPtth
occa1a1onal bryozaens There IIIIIIl8 IWIo lagII quantitibullbull of
~ retained -gat the algal fu-nt nPiate lIIIIIle alsO cleansed and stalJil1aad_ bllton ~
provided On 12haUrly~ ~dllrUght luar~w1 RddtUOlllll steel plat8s ~~In Alii ~dIxk~ lit
Uwrpaol (lie) IIIhare they IIIIIIl8 co~ by middot~Of~
(xtllus edulis) and the lISCld1M Clona intUn1l us
~1IIIIIl8 roved f the pJaqa NP~1id In cl_
ta tor 5 days at cansqnt ~ toIitDUampe
pH WId pOz ~1ane frc the extuMl t~~
the fouling to the Milstrata llllllre ncarded tt elecUadM
middotheld in a rigid pttllpex ~rr1et attechld to II ~ly
Jointed GIl on the lI1crcIIanipulAtor To nciad the muM the
tnraugh the foulJng tI1IIIIerds the ~ta JI_ta 0( 1 _
could be acCUllitaly controlled by talng the tIM clrlw of
the n1pJlator Iftn each lllc~ the DVIII ~oIdlft9 shy
allowed to _tabIU for ane wte prior to ncordJng the
VGltge~
~
bull a
-lt 1
l~to the_ batnacl~ 1ls~ COC)trol - fmiddotmiddotmiddot
recod4l9sof W pH 8I)d pOi at the SIJIIEdepth outside tOe 5hell middot-middotvmiddotmiddotmiddot_I~ lgt~middot middotmiddot~_middotmiddoti 1gt )
wete made
-flesolts ~
(a) pH IIIldIDz p~lllllS
The _re~ts for Hancl p02proflle tluough both algal -d - ~ gt Comiddotmiddot i -
eniIIIalfou11ng are gi~ in Figs Sto8bull --
TjpicalProtUeis daIiilttuougha t of Entronlorpha are ~1n -bull l J ltl ~~ -
Fig 5 - lIhllsttyp1cal d1innal fluctuations CICCUIJ8d during bull ~i bull bull e bull bull
photosynthetiC- II1d non-phOtinwnthet1c lods (Fig Sa) mad _~ d q _ _ __ _ i bull
decreases inpHlIIIIl1I~ IIIIa) fro-theaul-f1lCe of ttWt bullbull bull ~ bull
dcyenI tqwards the abstratacn tdch it enclOSlild (fig 511)
~fal1s1n pH--nted to as-ach as 2 units Over 9 IIIj
dIpth
figure Sa ahoid the thangea in pH -d Pl2 between 11ve barnacles
with depthln a stlat1c envirornent A steady decrease in ~
occurred as the plata IlUrfe~es toiereappro8ched pHWtially
fell on one profile before rislngbut lOSe steadily with daPth
on the second praflle SlIdlar changes are IIt10wI in fig 6b
with depth into the shells of dead barnacles 80th oxygen -d
pH levels were re~at1l1E1ly steadY ~ similar to the 8IIIblent - ~- lt
envlrOlWOt Until the -actual dead shell IIias entered When a
IIIUked drop in both fJ02 and pH CICQ11Tedbull
~llell in rigs rit JllellllUrellt$IIere maltle ~
iOlfillidual IlUs$els withtiRle (Fig 7dlb) In thll first
experisent elactrodes were placed indicated (fig)
arCUld indivicUllalsln genU flawJog ~ter The
resUlts show bull ~tle fIwrease ~ IID2 eMIS bull aiJc hour 1Ucj
The pH~t of the~ II4ttIn ~~atmiddot~
81 (te UlbientpH being 80) but 1GII8 ateIIfIU ~ the
1rtIa1ent aJphon to a6bull 111 iIadIIr ~~thI aff~ of
flOiW 00 local ~ciIndltJans~ the ~
~tadbut tile flow at~ turned Off tar 25i1aur
Jadin thelllddleofthlt ~The nailts g1R
in FlQIb -PD2 IIMtls mae ~y inflQldng CCnd1tJanamiddotbut
fell -zkadly fter alhoit lao w flaws caadawJnit a
~ fall lit 90 alii Levels rapidly ratond onca t IIIIUtchadDli agaJil No ILICh teed varlauon ~
in pH around the eldlalent 1pbn 1iIIflImI a taIdyJncn4IM -
obsmt8d aspec1elly cleM to thIIen1Ml CondltLni around
indivOJa1 ~ lIIhllst of interest are lMa repl_tative
of field conditions fro around groups or clIIIPS The
IlICpIIr1ants were ~ ~ted Wittdn denSe clUIIPs of ala
fouling a plate These r8SIJlts are given 10 figs tie (1Il1Ud
IlUlIS8ls) -d 8b (within byssal thrtiIds) In tatic ccinditJans
amprCUld the _15 u-a1(8$ ~ fellrapldly (45l) reelJ pft
rose sl1rjltly and then feU aftar 2 h llhen flowing condltiors
were resuraed after J hthe ~ rose to the original middotlevels shy
The pH rose lArlcedly at the eltictrodenear to the xhalent
bull bull bull bull bull
bull11
bullbullbullbull
it
I
8
s~ (3) but reriialned stlaadyahd ultilllateiy fell silghtiy (2)
inttie genetal fooling inassbull
~
III1thihthe1JyMli~~ Or~selci~5 IiIfieretilltti($trltus teniis ltO -=IiUlteliter~ ~ ~(Flg 8b) ~ lIIithrrolaw~ cCincentreuoos diCIppea framp1~D lBigtif3S
~bUt toaitnpl~lylt4i 150 IIifIo lIIhfjnficw t8st0ted~middot pH
t wi~1rithi thread ClUIps~lMl1BtO 74 (3) IinIt B~ito
785(2) lri-staUc cinfit1at IoihUst ettn88dgtiofthlt ~
it -flnedf81r1y~at aroOOd17S~ lIIlanf1Dlilwu
rellitoteilpft inltleUyJose (by 04 UI1ts at Ii1ta3) but it theo
bull fall only to startrfs1riq tgi1nattM not theIlXper1ilent~middot
bull Dlseusslont f tMresulblmiddot obta1nIid it ~be cl8lirly sean that f(IIJUngI
bull species vethe abUlty to Gtly IIOdlfy thdr loclmiddotmiddot
bull InIl~ts
bullbull L1t1t end dark period tluctuat1ona in algalts IBre obsarved
end ware lri close wpeeIIIen~ with those reported by Terry end
bull ~
Edyv- (1981) with 1ncnasaln both pH end IID2 llIrJngllttt
traquor1ods lIhilst thlshappened ttM surface of the t pre
bull fU_ts ware frelyprojecting into UJe water -deep intottie
mat nearer to the solid _stratlli whl1lre shading fll1lli light andbull accuIIJlatlonmiddot of deconIposJng ~gallIIBtllrial DCCIINed pH valuesbull fell IIIBrkedly Concurrent wlth this dlQp in pH ilaS II fell inbull
bullbull
9
oxygen Such t~ral and $PiItial va11ations in ~andpft
would certainly qive 11se to corrosion cells middotatmiddot the sUbsttatie
surface ~ly resulting in iorroaien pitting
The r] _tabellc ctivlUee or fau)Jng -=raJrwerttmlbullbull
together with their prapeN1ty to-=ci-1tritla~
tlwlsalves wIll also lIied to 1dcIoenv~ lIOdifktIona
Resplretory activities wU1 tend to ~~ tn-the tft and Input caapanants auch a cinia~nY qatk
1mIertabJatesmiddot ara mlk 1bullbullbull middot the bulk of their nlUagllnoua
wastes ~ excrated as ~ 1heexpftunt ncord~ pDz
end pH cningS amN 1nd1v1Q8l~ ~ ~ri
claady 1nd1cat41 the ~ of _tft~t GIl tt ~ of lnIirollliental changes Inmiddot expr_~ta lllhere no Wlter
--nts occurradthmi a np1d dacreUfi1ll pDz
COIJIdalo be a ~decreae in pH (no a) v fladrlg
condlt1C1ns are r-ed tIwn Ieymiddotmiddotor iIay nat be a 1bIIeIqI1int PH
1nctease dBpwIding ~ the --- ofmiddot iheltr ~ by the
organl1n the cluIIp 0wIgn wui a influenced by the
proxJa1ty of 1nhalent or ~a~ the dlnsity IIId the
tIlJcknes$ of the clullp w111alsa t)IMt wiuence in ~
wetr -N_tbull Hencein thick clampllP aUt and detritus
cCUlllulatlon is are COIIIIiIn ~ theh billa IIIdfbls
restricted Br8IJIIltdow1 gf organtc datrital _ter1als CI3IIIIUIIS
oxygen resulting in recLlcing conii1Uons llleasurlllenta haw
shown that sulphids qulckly build up In such deposits and
sulphate recLlcing bacteria y tie-~ SRBconcentrtlonmiddotin
J -I
II II IJI Ii ill
II
-bull I
11
10
the depOSlts woUld terid to Ile lOW at We start of ~r1ments as
condit1onswere nat favourable (POstgate 1979) The cantlooQUs
prockIctiPn of r1th f~plieUdofeacal_te1alor IlUlpllonill1l
~ts bYpoundilt8~ (~~Iick1ngand 1IkIod1) hOweva ~ t H 1 (
IIIOUld rep1dly altAIr theli11l~t and the rua~ofSRB re
notedtofncreasemiddotttcllll ( 1 SRaIll1taat llaUt 1 x 101 SMIIl-1
aftet bull -th
IUctotJlIIl corrosion canfe=t _ta1sln a1Ullberof Ways
(Iverson1972 fUlllIt end King 1975f 1Uilr 1981) pH can
aff~ the ectivity of the 1IIin organlasinuolved the H 1IIho
hava tin qlu pH range beban 10 12 (Graff 1981)
although tIdS -can be lnf1u8nCed by ather fllltors In atI1tl11n
they lIKIJlre a catbon end nitrogen aurCa 11 various
salts tin alactron accaptor (HedJart ~ idbullbull 1985)bullAil of
U- facton are availabll in the debris -alltting b41tween
ar1na foullng -d the 1lUbstra~ 110 providing bull potantlally
corrosiveanvirorwent
Evan IIIhete such accullIUlatiori does not take place varlable
changas inoxYQ8ll and pH ~trations acrossthe suface of
teel soch as C3I becaustd by foul~ ~ C3I give ti_
to corms1an calla lalildlng to piftlng corrosion
There is an initial tllld8llCY for the pH to rise as a relUlt of
the excretoryactivi1i~ of fouling aniIIal and this 1amp UIIllkely
to give rise to corros1an problems Eventually ~er feacal
h
1
~I
materials and detrltus~ill build up around or~anisms ~re bo~
low pH and p02 devul9ls providing a potentially coJJOampiIu
envinx-nt flaw does dify thh _build UfJ but inmiddotthbull
Ixparlunts carrllldotttid notprlvl(lt it R~~r it
l8Iilated the depth of deposit ~1~~ in t- ~ It ahoutd alao r~~- in f~ population so-1QIJvlWela 111111 be dying or dUd Thlb
~lng~wUl~1Ielacal1Md conoa1ve ~iccandltiorJs-t SR8a 111111 b4I fCllllfC314lng ~y pOIi1desa hlbitt~ HtInd 1tt~ WIi~ul create 1~1Mct Jrcx-ntal dlffarenca ~ ~th l1kel1haOd or pitting
n
References
Albilnder J~T 1 bL~1953~~(9EthII prockIctiPn of IdcroaltJodIIs UIIId in I~ studies Rev Sc1 lnstra ~ 528-531
_r1can Ptbl1c Health laSoc1ation 1975 5taIIdIrd~for the lIIIIIlystS of _tars end taatms 11lth pounddIt1on 1IPHIi-~ 1015 ~taenthSt NW 1IIa-l~~OC 2IJ]J6 USA
Bau-8laquo=k1ng C~IIIG amp 1IIoOd EJF1955 Biologlcal ~ in theestuarlna envi~t 1 EcolClil) of the ~ cycle Kcnlridlkjl Ned Akad -t ~ Ser B~ fi 160-181
cter DB amp SUver IA 1961 In IRafaranctlElectrodl$l EdltolS Iva DlG and l-z Gl lc8dilll1c Pr London Maw Yodlt Chapter 11 ~
Davies PIII I 8r1ric middotF 1942JII1c1oel4iitrodas for-aln local oxygen tension in n11 tissues Rlv Sci InStruII Yo 524-533
EdyileariRJGJ ~ Tetty LA UJ83 The 1nfluenc ~lIlcro algae on corrosion ofstructural steels used 4l~ the 1tIirth
Sea 81odeterioratian sect 336-347
bull bull
~
bull - -- ~It Jf bull
- ~ 13
12
Terry LA I~Ed)VeangtRfiJ 1981 lIi(toal~IOI1CGrmianF~ l)i 1St1oJhemailnEifooiiiigffixed gftshoreIt Bot 1Iar ~ 117-183 lsect~~jlaUoQ1l1 _ ~t Ef)e~gy Offs~r1I Te~lOtjjy Paj)8r
-middotltcntpI)~it$t~l91-16PP~middotmiddotdl ( I
bullbull
iIIalk e1 JL 1971 bull Ion specfie Uquid I~~ e)(charig~ ndcroelactroclels Anal ChIIaI ~8S-Q2 AGraf1~ w~S 9S1~ 1orrosloh InFIntroooctI00 tooffshOle
51~weSoes~~fab~lcat~onIns~allat1on Gulf Wall PI aelUnger EG 1982 The appl1~tlon 9fMl1ikIngmiddotHoust(jfW~Q23g-258middot icroelectrade in the study of pH end PD2 in lIHinli
HerbierC SN fGH~rt~ )RI -Stoctcdalpii Watltif1Sal RS ~1ronIIInb Ub~~ 27l-Z79 1985 Fac~or lontro1l11)9 tha activity ofmiddot sulpMte
bull Wall P 1983 The appllcatlon Md ~ ofiwrJ)e~c~odl shy-taicfii9 tiiCtetili tn~ diiilncj tar infection
techniques for the iiaurnt cif icraeov1ror1liltital5QiCiety ofJebo1- Engi~ SPE 139781-10 _i~~ - ~ ~ i ( _ IKIdif1catlons caued by aGII8 rlnefou~lng -C~ebullbull
PhD lhaaia Ibiverslty or ~tar UKbull bullbull bull -
IIH1nke JA 19fiSa In EXperllRants in Physiology and middotampioClIiIiiilatJift l c E(If(orltGAiKetkutgt ~c-Presa-
Wooltdngton AObullbull Davenport J 1983 pH lind IlIl2 IMla ffl~- j~t~5 -lMIIleath 1IH1na ro-fauUngGqWl J bull Expbull 1
B101 Earlsectsect 113-124 Klnke JAIII 1969b II) Glass IUcroelectJQdes poundx1tors Uwal1~Ui~ OrOllt arid~t Nt JOhn fII11ey YOI1lt thaptar 11
~-f _f~ ~ ~~nt ~ Igt
bull Iverson IIIP 1972 Biolog~ Qrroaian In Advencesin
corrosion scianca and tecIYIology 2EdS FCntaia G~ I bull IIS~et RIIIbull p~ PJaSsLancIoIl 1-42
bull
Jorgensen BB Reusbech NP I Cohen T 19B3 Photosynthesis and strUcture of bIIn~ II1crobWp MC1OIIlectrocki and ~ stud1ea of four cyanabacledalt -~usllWtOl ~bullbull 1~f093
t l~ O~ ~~~ H~ ~9s7~c-gtHatste~iecfn1k van shy pellediftlrtenPt - Stlchelektraden (1-Su
AiJBitiKlificfiliilislflilr)zlIr polrapusdiailJIIissui1g_ liIserstoff(jruQkas fur d1eB lIsUbullbullung der IIliJlttOUf1cillatlon PflftgaHArdI ~fl39
bullbull Mller JOA 1981 tals In EconcinI1c II1croblolooy sect McNbUdbicldatariOtationEd Rosebull A middotAcadelc Praulondcn149-202
t Mller J DAI King RA 1975 8io$~iorat1on of lllltals In lIIi~1al ClflPlCts of the deterloration of tarWbullbull EdsGilbertRJ~ middotloveloCk 0111 bullAcadeic- Presst IILandon 8Jo103
bullt
IPotgate JR 1976 The sulphate-reducingbacteda
Callbridge tIn1versity Press Cliilbrldg8U-K
Reusbecb fUI i Ward DIII 1964 i~~~1Ida5tud1el of
bullt inter$Utilll IIIItez dttm1stry and photosynthetk activity
in ilflaI spring Idcrob1altAppl~EIi~ MerobW tsect4IlU-27lt I
IIbullt bullt
bullbull Ibullt
1
Inmiddot middotmiddot11middotmiddot f~~ a~ -tmiddot bull J~f
I~ W
i5iImiddot J~a t
ilmiddotmiddotmiddot I middotIt Ii
l
~
middotmiddot~~middotlL= g r
Lj Ic
t
i ~
I
gi II cEi ~ I I
1 t rW
D
tR bull
bull t 10
11
r
i
I ~
j
~
I
~bullbullbullbullbullbullbullbullbullbullbull
LV
FigS pH and p02 profiles into macrofoltling eomlllllllitilS dominated by Balm trenatU$ on steel plate 11 two pH profiles Ib) two lIOai profiles leI pH and P02 profiles into dead barllllCle shiella withdetri~Us I I
itIbl r p02 ~ I
nvnHg 11 70
j ifJi eo 1amp0
laquof it 30 s
I 20 N
ca 11 10 a o -- shy i bull
11
80
19
781 o 2 6 8 10 12 0 2 6 amiddot 10 12 dIpdI Qf ~iftto fouling In HighIst numbIII pIatIi
8deg~~
~i c5 iioo5 fiil
pH 8 bull_ ~
~~ ~
_ bullbullbull- pH
-pOz
(cJ
e-Clf ( j~JJlaquo1- --~I ~a ilil - _ I i ]1
~ ri en i ~ oJt ~iJ id tUI
2u1U I shyI t
shy1 2 3 6 6 7 8 9 10 11 12 i shyDepth 9f electrode (mm) into fouling
1 i
1
~----------~-------
~
~-~ lt1~
middotmiddotPmiddot ~
I
V
~
ill
1 Tbellitt~~ of cathodiC protection OO~~lIic~tlO aridrfOflllUlc
ofi AA~Uoullr14l ~1~I
I
ael~lner EG middotIUtcb8l1AJ Dept lt4 lInY1roMe1lal Bl01011
=~~venilt7 IlariCheW 1f139PL $ ~11al1df
r
I
Ablitrict ~~~ e~l billttI ~ted with ~pert1n ant11_1111ampp paint llere itpoii~1n _vatI- In the tl~ld iUIit 1bOioatbrt~ScM Were
1IUbjll~ to qtbocJ~C ~teltt1on byen _ current (~)SJi inci EDAX Iieamp~ta IJdlctecI ~ lachln teoc copper OIl tbePlCtec~ pa1n~ atter14 dQ1a~ protected Pllnt abo ebowed
) reduCed bacteiial activIty butmiddot ~e4 dltoe Iu1d protOzoanmiddot co~~l~tlont~1l IlU l~byenCOCltact not byen dUfwIl00 ~r _baRl lrillolvedln tOXln nil ereio noted
l ~~
1
I
Introduction
Applications of protective coatings and cathodic protect1on(cp~l
are recognised corrosion control methods for IIIBrine steel Itructureso Cathod~c protectlOll preventl conCIsion by Int~lng til8ctrl~l
currentS trem externalllClUfCa to e~~tthlJno~ e~ carrollon feetlans C04U lUChu tUauJ1ng Aelnts a
(
barrIer to ~ the flao of couos1a1 ~aboIHnu~r
Dccurrlng anodes cathodes or wlwit gelvWc CCIIII1 ( 19681
flinger 1980) AnUfauling ~ also JJICOqIOrII~ b1oc~~
Tr~ylt1n nuor~ (~) end o~ Oxide (~) (~ flJ1J
- of u blacld1111~
V _Ut Evans 1981) ~ ~
end growth of bipfoullng 0I1I)IIIII1IIIIe Wdd~ ca ~ of
paint rusendmiddotmanemiddot ~(r~ til )
As corrosian ~t1on ~ bICCIIIII -an ~1tlQd ~ of
antiroullnQs 1 conjunction _ith CP have bllClIOr ~
resultIng in reseudt 1rwestllj111tlng trw COIIIPIitlbUlty or VU~ ~t t _~
rOratlation to dlfrert levI ot CP (~Eelf 18110 ~
~t 1980) RelatIvely 1nIestiljlllUans have ~ urtw ~
or CP an thIt anUraullngpert~ of palnta
In thIt preseot 1nlIestioaUon tV~iln111 ronutlon ud
cOIIIHrclallY incanjlntlon wIth CP was ~t4IC~ Iw I sraed current (-966 l1li) arid exposed at the 1 Strait IInQlbullbullbully
~t1ons weremiddot 00 the IIhcIrt bnifects the PprIUd_
had an a) the various COIIIPQI1eIIts of n pe1ntf1l11 end b) l1oft~
developaentbull
bull bull ~
~ ~ - f
bull ~ _T~~-middot~ ~~- ~r _ -$1
(1) ~rlttionor Painted Steel Billets middot_ ~~tmiddot_tlJ middot~hmiddot~--~fmiddot ~ middot~A~-~~middot - -~ ~
Thirty-al ateel blllets (8-10aD2 end l depth) were cut frollbullt ~middot~middot~middotmiddot~middottrSmiddott ~jS-~ It bull gt~ tfl - middot--imiddotmiddot ~ t1dItrd steal plates The dimenSlonsof -thB butits-lim chosento
bull -~~ middotmiddotgt2h ~ ~-Imiddot ~~rt ~- -middoti~~ ni ~~middot1 l middot-~t~~middot _
(a) fecilltate amiddot Ie detrucUve -thxI of prePraUonpdor to ~~_gt gt~ k7i ~_ t~ 1--1- _ ) - +--_~ -_~bull ming electron ~ 4II1CI (iialloWdirectobs8iVatlCnS of the
- i lt-- ~ gtbull -o1e billet 8Uface
gt r~middotmiddotj ~L(~middot~middot rmiddot tmiddot J ~ - ~ - lt _ bull Single non-standard (1hIula~) nickel lIIireSmiddot Spot Welded e1Cing
bull ~1deJ ~ b~i~ ~j~~ tri afl~1b1ipOi~ f ~1t1 ~4middotmiddot f-gt -~ ~S-- ( -- - - _bull resin EactI billet then Undb~ tI1d prtiladlllltha5O __ eyilr
bull ~~_ ~~imiddot ~1iil h~ intlfcili~ ~eonbt1nlyi Tr1butylt1n F~id(18n)~~~O~dettu20i~blcicld8is 1IIfii bull i - ~ ( then 1IPI)l1ed to glvaadry Ulil tIi1c1ltnOss or 1(1)-150bullbull
tbull (2) IipreSSeCi Current Syat
bull A rigid ~tic due 1I8S drWedllliihwrtv-s41 liiIihOieS IIPProX~
bullbull 05-10 cia episrt ~ Pilnted ampUl8t Wsfbed IntaPQSIUon ampy
threadlng the nickel IIIlr8throu11 the dinto a tarproot junction
box Eliltilen of th8~1l1ii (recelving ~t-d curnritlere IIIlred
Into bull block connectOrrecelvlng dc trOll is potentiOstat The
rainJng iontroi blUets Wet8fbedfl11i1iiy lIIithln thejtlytlonbox
i~ityd~c IkiliPly ~~ ~lat1ruaWiode 8ncIacalOliel
refer~ electtodetoiere fllied ~ the per1aetat of the plut1cdlc IIIId
connected to the pot8ritlostat The desl~ of the potentlosbt ailoiolEld
~PlDteItlO1TClJrnnt to be MIllntained at 8prampset levtl of 966 II1II
c
r~
Grid raf 1J5s-t 114iH 563724 dud 0 Juna
Alolecule andbechr1a in general are ~ to
sutface lI1eroleyer (nlUSton) of _ter (~bullbull 191KJ)~To avold
contaJnatlonof ttw painted surf by U- -vents
blllets Were s-nact in a 1 conte1ner or atc1le (02i ) f~~nd
ter Iih1ch then f9ICI in tv 1IIhere the b~
be r-oved undeNeter El~~ bllleta fa Mcb trtai bullbullel
removed over a 1 dtq (336 hr~ expoeure perioda
each being ~~bn uposurnof 1 5 11 iA 168and DlIIauz1r
Arter field axpauHli the billets wan
fUtered _tar to ~ 1_ debtlflMd far 10 aJnuta ~
wv ~~~endattid sdbUiUd(otO
~ ~l f~ nn1ng el~~ --
lOOted onto 1middotCIi ~bullbullWIsend carbonmiddotcoa~nmiddotmiddot~middot f
each billet Ws~ Iis1ng a ccr1dge ~el~~
(5360) IIIlth anergy dJ8perslve X-ray alcroenelre1a (mx)
_lysis the les Wen reNed end coated III1th vold to al]Ql En
detailed sat of the alaquofa1l
(4) Lfiborattux ~
Painted taa~ bUletalllllre pnparad ~
in a Mrlne reclrculet1ng flOlll _ ~
f1Oll the ~ ~a1t~y Tha~~Un bullybullbullbull
10C M1nta1ned ~~t1as of ~t~la protazOllbull~bull10shy
(Entermap) ~ ~~~middotNlt~Eamp1~ __~and ~ r - ~ -
certaJn ~lc 1IQ1~ 1(1 the
the painted
~
thre repl1catft fn
plKed 1nto(022_)
- thIine1J~~ -lt bull
Atter mAX
(1-2) and i-n-t bn collected
14OC
Pain~
-
12 daY~ exposure and prepared as in d)~ - t
~I ~~~-- i~yen 1~ v
Rflltsi ~ _ ~f-~lI ft j bull _
rt~ll0~~~llY~~~~peln~stetsl~11i~ex~sed for
y~t1~J~ ~~~~1~1~~1UII~ya~kooJ~~~lt ~ ~~_~~I1J~ ~~~tomAX -lysis ~era than gold ~ bull bullbullbull c - _ bull lt
~~fllr ~~~ vllUll ~t1ans of the rfee htures and gt Co
~f~ - ~~~ ~f ~ ~aobta1Mdue IjIll811 iI1Flg 1~2 The IIIIdn ~~ ~ lt)ji9 )tmiddotmiddot - ht
- bull
~I rIIplt~ tIt8 -~ cOlllponanta of the pei1t 1bullbull WelIIlItr1x $ilj~t-~~middot - ~ _ -_
-IUch PVCbQed end rap~13y the chlorlna peak the
i~~~-~~-~V~~~-t18 ~~~~ ~~ Th8
ral~~i~ JlrQP(lrUo1sof ch 1Il-ntal paek~ll1d l~t_ Q~7 -gt- ~ _~ -- gt ) gt -_ -lt ~~ the 0 -end _ xprtsntabullTIwsa changes are -r1sed ~t~ lt~ _ ~ 1nT~ IThe8llia$lb npresentthe chengelI 1n~t1V8 p~lons
ti~~~ [ ~gt 3 - - - _ - _ -
Qf nellllllllt within We surface layars of the pe1ntmiddot ru The
~f~~thtJn -d ~rfeli avr the 336 h11cxI ampScanbe
SII4iIl r~ fig 3 al~ tht rta of~ dirfers tJetwaeo thaa ltmiddot~1- 1ft~- ~ _
IJOl appfiai-ed to ~1nlt1elly until ftar day 168 it fall
~~l~ 1h8relllllrelgn1fi~ diff~be~ qrOtactlid and
proieCted (at -966 ) piIIllnts tor lioo COPPer but not folimiddottin
Ibi~tlons Of the Pain -urface 8hoWad _Pil ~ witilt1M
(Plete I to IV)~ 1hePllat1na (~) pelnt(Phta x and b)
Clearl~st-i~~r alots on the SurfllC8 IiII1kh~ter1sed ~- f hlt1 lriitUl tin conCantrat1on~ lllariy slotsre Partially
~OIIe~ ~ tile _tr~tedal middot1lere rs ~an~ densely packed
grerltr~ ~th 1ar~(iIa1niy cotifIIr) 8ntt -ampniBil (lIB1rily 1rori)being
middot vl~l~~~~~ Dtt~i~~~t~ r~ ~f~~ I~t seawatllr for 11 n IIIOre slots appeared and th~ ~~ess _trb cwer
--if -- V~ c~~ f(i ~~~i~
(Plate Ie to It)~ the granules dld not protrude from the~rtace in ~ ~~~~j1~r~-ffJmiddot0~~
suhap~~r Both 0 nd --960 at ~~pa1nts ~ - fr ~~~~-w1-U~ ~~t
sl~llrat thi stag~ (Plates Ic-d II1d xrnspac~IlYh middotmiddotWith middot middotaJ~cmiddot gt incraallO ~alolre t~ difrrqcbullbull middotbatIiI8IImiddot the protected fjd
bull ~ ~ ~t~
IIllrotacbldpaint ~~lIfIt~ftitr~ h alQt~ p~t bull u bullbullbull J ~~i~~ 1~~
lIIith IIIuper lIdguon u 4lrotatad piIJnt (Pl8ti1 II) 1hD1II8S
Ill )1IMdw1lillS gnnJla~on f-r~~ dpert1cl ~ - - - ~~r ~t
Onthe 966 1IIpetnt t-var tIIO typeaofan with diffannt c t
~hIId~(Plate UI a to c) One hIId tIII-1 bull bullbull e _ bull
fUll wJ1hleas abvlaus alats the o1fwl bed C18UU1IlOt ~litUi middotC c ~ ~
less ~ t11 h Gnnulatlan __ but ~lIiIZly def1nlld in
middot~ latter Afte fila bed tiIIrtIId 10 ~are ~~j(
extansll(tlly ~ the~tIId(O at) tIIIIple lIIIdngthe~ - ~
middot orarallet1ai leas claaI (Plate I~) f u II1ao CQ1tJAJ11d to ~Gn - -
the -966 at 1 (Plata III d tor but 1JI8V8n 1nittNdcneabull -
Althatqa -middotslots wrs abvlaus becaI1ng 1 cwampr aiI- the orwWat1an8
f1nailyftar 14 day exposure ~ fUll drMIlapIIIent on all la - ~
are extensive with the 0 lhawlng both alota pore ~
OVIKgrOloln (Plata II c to f) Gnrulatlon 1bl to ~ gt~t~~ i
or the th1cknats of tho ma ~the ~ at ~ (~~~~ themiddot - - _ middott ~ ~ ~ - -1 1 t
middot f11bull IIIIIS IIOre extans1 thIIIn at dIlya but IIIIIIIY bull1Q~ tI st11l ~w -middotr~ 1gt ~1 ~
obvious fIIany ~ loI8re still p~ yen~~~~~~~r
than before Poiu appearedto be f~lf~ila~~c-~~~a (Plate IVf~ label 1) IIIIl1ch broke~i(2)~ ~~~~
t t
bull ~lS (3) ~ rflC~~ ~i~t~~l~~KI~ t~ldiIiiicyul~loIiih ~~Rht ~ )~- -~ - ~middotd 40 ( gt ~gt ~d lt1 t ~ Jt ~remiddot4amp tha paint trix cavity bottom eroded away reVe81~ng ore
t ~~t~ por~~ ~l~ P~]Y p~~8dby di~lutiaA~ ~~~~ti~Jr~gt~ta(l~~Siinll1cl1Ud~f iUiIiI reriatlGllbullt
bull a~~ ba~tid colCriAtloriccic~ GIl ~~~d ri -966 - ib~~~amp catQn1iti~~t~I~~(~t an ( ~
t the ~~~palnts (PIa_taiiI d~~h ~wh ~ n td have ~ CamplluJMitiIVc) ~~middottti o~lgtnalrf tlt of hIght
~ l=~A ~J~rW tYtles~ p~fiiilriQcOCCold to~Ja to jU-t ~jmiddoti~ai dlatClilS ~ir -jlati ~tbullbullent aNit ~ t ~~ th8middotf~ld~ies~~t~on~ ~ -966 prQ~~~(P14ta Iii~)Omiddotiliof~m1hiiHAiiipieat ere~ll-(piitir~ p-iot~iilans(PiA-iII~ end c) nbullbulltiledt dl~by1i~~ end wirinali1sheii ~cMI~bY thI tili
bull In t-~~ ~1i1nt 1oIIIeHfd~ ~m1IS (15OClind a t ri~r-dlatoa ~lor P~t dlataiOlonlt1anGIl ttw-966
t
bull ~
pr~t~ ~is~ih iion obitJauIJ A ~rot -=1 ~ ~ i __
reprasenttild on the 966 painted 1iUrfc1ncluicunljj ~ rwt
Achnanths Colonisation by bacteria 1n the laboretory bsed
bull eliperlMnt tollMct sbiUar pattern to the fild ~
bull bull Dilicueii
bull bull DUt 1Iidnt ~lona Mve dlttllrint ~~ ictian
It ~ JntM l1tenture ttt Iff yen __ JIll the
bull ~ bicins ani 111 ~t INchlnil (PhillIPs 1t73t EWene~ 1981
In ci1ntlHt tJWY ~tit ~Wl1c iIuCh as 1rbiltyl tirI
~ lite wAlII]ltiClr dlsP8tfu~ middotthiirsttilll~intaJn~ a
bull ~
lIIifoflll ampalCentraUon through the filII 10 thoIt biocide rel_ at tha
paint surfce 1fI$Jlts in IIIOre ditfusing ~s to repla~lj -iflJ~it
resulting 111 bullbulltNdy lr leechlng rata M1llat this IIIIY be middottruIi ot
tin ~ it was nOt true ~ ttw middotlBlf fOZWJlatiGn ua ~ bull - i - bull 1(1
ObSfvt1onundr SE clarly ~ tM t1l1 to be present
~ly lIS OIIIdle-llk parUc1~~t tn lUlfiice (Ito
IV) bUt usa ~ the dIpth attt fU-middot ~y~
If one considra the concn~rtlonmiddot chMge 111 1n ~111c COKICIl8rlts fro- the paint tU- i bullbullbull tMWei b~ ~ Si6t arid
tI-~t iron oxidt 1~ can bt (Fl_ 3 IrId TIibl8I) ~t ~ ase cona1derllble d1fftrenC8s ~ a Un decz bcdYMr ttw p1iod IIIIwraS lron~- an unt Ught inCr h ~ incqJper ~tlGn occUrnd _idly at ttw ~~ Gfb ~ period ~ out ttwn fu nPldly ampQII1n ~ umiddotirid II phast This IIIOUld bt tIlq)8Ctad It cantact l-=h1ng we occun1ng
IIltfIOS8d pertlcl of copper would lMch rapldly on t-n1on 1n - shy JIsewtr There would thlll1 ba bull J-a a1iltUy leu avalUba
i1 ~ ~
_tedal becfM uposed Thi tIOUld ttwn IIllCh Thla pulSing efttct
IIIOUld be expctad to -oth OIItlllith U- putic1 __ -lllible II tot 1~ GIl a on r-x _lao Thla teet 01 put1c1e 1tUng
can tit GIl the lIUrlace ClIlC8 it ~ a pan 1amp t
a 11aM bet in Plat I to IV IIaIrIg _ lit la_ II ~ 0- ~s _- PIate IW $llldl ~bull nshyot poRI flllNltttn Al~ tin ~ lou rete thtn I bullbullbull atill Ivwnce of bull al~t pulsed 1Itue rrlict tII1IIWd
elqlCtaG inIl ttwauUr ~ I I I
0
~ i
~ ~
~I J
bullbullt bullbullbullbullt bull bull ~
bull bull
I
bull t
~~
The same seqiHllle of cevity follliltloii (intttecaseofthetestjjltle
slot- shaped) fClliOileciby SecOlld8rYtlolesfotlllingin thll cavity
bottollis releasing further deep8rtox1ils (Plate XV d and g) As
cavities_re ProdICed ~ tiecaaIelnvadedby_ter~ onelilauld
expect 1ncreaa 1n1IOdha and dIladne lMls t the t1met
which the toxlM uw ~ Ths fCMld tCtHi eo (Flvmiddot 3)
~rttno the vi that_tat ~t1on otthe~ ~lY
il1Cl1l8S8S Idth UIiJ
Differences f~in toxin li~~to Ini 966 ill 8811P~ F ~ bOth Un end ~ lcMIr levels afillW
IIIIIlfI ptaInt(~1~~1ntha~~ ~~f~~ cst rklld with tbiIccpper ElJIc~l ta~Uona invalvinv
w pl8cId at a~ht~~middot3lO~lth Un cqJpel bull
(-960)10 ttW ~af this expei1tIIri1ts thIsiorelfJltdt -
that cqlp8r b8IiIId ~ IIiIOUldProc-ln f41ip1dly t thellllgllitiv
patentW _lied () as 1t 11 further f~1t ~ witt Un
III11ch is u-tat aqulllbd Idth
it If ttda 18 eo~ ~
~ )
frOll1 paintscould bl anqnced by an illprd current clithodic
protecUansysteipos1bly ahart_nino te 11fe of the paInts
antifouling ability Bacterial coloni Uon IIIU rtdUCed on the -s66 paint This
could have been due to unflSllOUlllble electrostatic cqrge on th surtece Th1a effect CIIl pley en IIIportant role in the t~of
bacteria to alrraces (lIIarsMll19E1O) As bacteria appear to IhiJw
greater toxicltyto copper (DMpsey 1981) it is Klre likely that
enhanced cqJper leacll1nl On the -966 AIIIPl c8used the recluCed
activity The IDlol8r toxicity ofUn to bacteria (as reported by
~ -
DelIIPsey Hle1gt or its lowes leaching rat was sUpPorted bythl
relatively rapid recolonisation of the slots by lIIicroorganiSlllS ~ftar thE inlt1al aurge of leachi~ Thailllpressecl current dto ~~ the oppoSite _ff~middot ~r on dtta colonstJon l1lI-1- in the
JIbcJrtory~ta larger rUDIIra f~ fit pro~td ~ther than uriprotected Pafnta
Ccnlus1one
1 tiild-a 18 presaIIUd ~ thattJn~ ~~
~llNChng notd1rf~ ~
2 TIleuseofCP~ ~CUIftnt(at) on1h1a ~fca41ng ~- ~
pa1I]t~ to ~_lAIIach1ngn_ at toxiN - -
3 leIJstaPidbectar1al ca1on1aatJon acaampmId an ~bId Pt1ntJ ~
4 JItore larger alcrofauJing (Oiat_ and PzOtoaoana) Gr~_
occurred on the protcbld~
I l
I
bull bull ~
mfEflpoundaS middot~-t9i _jgt- ~ _ _ ~sev M~J~ (1981) (cilonJzatiooof alUf~lng lnts by _rine
Jbull ~teda9g~1ta IIarina XXlV~ 185-191
- - gt 0 Ie 1 _ - _
poundV8I$i I~lf (1~P~h 1II8rin~Ugae and Foul lng AReviw ith
t p~l-i9Jrlr ~iiMdilto5hiP-fQUllfCiH 8ot~ ~r~ ~~~ XXIVbull lIPwl~J11A
Fotteth CN~R ~kk8nGB IWPIR1seJCiiiroaic1lihd ~e ~JIIth CII Offshore St1ucturu ElllS ~tjI~f~1Dt19i4
- - ~
UKC (1976) I~races ill ~W Ecology KIuverd Univbullbull middotraabullf~iddge ~bull C~~jII illt)Ia(t RC (1980) totinllsa Cattod1CPrOtect1cinbull middotmiddot8AtEmiddot~6haOCIlfif~ T~ p No 108 ~ 18 ~ 1IIHf bull
t bull lt ~J~~~ __ ~
Norkr4irulB(198D) Advanculn UcrOfj~lpound~ltiOyVol 4 (AlItxlaquoIder fl Ed) PlerIw Press lcxIdon~ lIP~ 51-85 ~ l~l - bull bull
PJijllt~i At(fm)~mr~~JnI~1fqul1nQ JlalntlJ Riumh ~JnOrpnic toatJngs 2 192middot
)~J gt0 c ~lt -- ~ __ RogerS~ Ut (1968) ina ~(lnt~fbItliP4ili GI COnoslon ~a Technology)bull bull
Sl~iorr~ VP~Ranald~ Rctd~~ (1S80)_ ~ Experllattal Stud1aiRelate middotEffect or Cathod1c Protict1Oi withmiddot terta1nr1c
Coating Syst~ 12th Annualofr T8Id1na1ogy Co1fItHnC8 liauston TIIlt p 9-12
j
fIG1 sPct FromMon IT8TJCuZOI Int Lliu EOAX
FlG~ Fro TlTCu20Ar-4 Dy Field Ellposult I~ir
il
I c
R hi -00amp=7
-I I I I I I I I I f
iii
bull bull
bullbullbull
I J
J
I Figur 3 Thl chan In reltl rqpoIQnlof -ariOtimiddotjOmponents of the paint film with time Thl vtic~1 ampkiS unl till _n ofmiddotmt Illment per unit OJ fe 50all Cl Na
Cu StI Feeshy
PlistinllFe S7~J6 704 3~74 908
f
r e ~ ee~
middotie ~lt - ~ shybullt bull ~
t -~ l Cu~ o ~ f
t --- 3igt - bull 0 J Sn
cc
t ~
~
bullo
~ bull e
bullbullbullbull bull ~
bull ebull a QI 1- bull 0 40 431282 316
101 dmllIII)bullt bullbullbullt
hr 0 ~~bulla 5~16 ~~ -~~ i166 4S i83 8~1B
24tu 6~480 SD~83
111middot 1~1 OS6
~ 5114 6~69 -51 UIamp middot016- lt ~rmiddot
1d o Mea 818 21 021
~Z21 199 1~1Qmiddot ~Jgt2 058
14 d o ~~ 804 Q~B8~~~a~~t~middot 685
866 372amp 8amp4 OS _i~11[ ~~64
Tttile 1 A81UvlL pzQJJortlcw 0 vxl~oI ~~bull Jn 1I1Dtitctel lt-1166) end lqIlDteCt6 (O)middot~af~_~middot_lodEDf~ middotmiddotmiddotmiddotmiddotmiddotmiddotmiddotmiddotc
~ ~
_----I~
J
I bull
I
J bull tmiddot
7o CONQRESOIMTERNACIONAL
I D~CORRO$IOr(IAAINAEmiddot~USTACIONES~v c)
t 7thflfERHATlONAL CONGRESS
ON IARINE CORROSION AND fO(JUNG
~CONGRE$INtERNATIONAL DE~CORR~ON ~E ET QES~SAUSSURE$
i t
~ ~
bullt
t t
UNIVERSIDAD PourtcNlCAVollncio7J1 NiwiemIif~ 1988
(SPAN
SECcl6lfn
BiOIogro marino Morine biologY ~1i1orine
Theunpub1ishedJllBIlu8criptEl have been reprod~d asr~ielved from the
authors The SCientiUc COlIIIBittee take not respoi~ulbll~~Y for any shy
error or omssi9n
Laa ponenciasno han side publicadas y estanreproducidas tal y como
sellan recibido demiddot BUSmiddot autorea pound1 Conte Cientfico middotno se responsemiddot
bil1za de cualquier error ~ om1si~n
bullbull _ bullbull_ bullbull ___ bull $_ _ L _ - _ --- -__
SESSION II Marine biology
Friday 11th November
SURFACE PROPERTIES ON NONmiddotTOXIC ANTIFOULING PAINT FILM
K Nanishi M Murase Y Yonehara M Kishihara and T Hirama
MICROFOULING ON ALUMINIUM PANELS PLACED IN THE TROPICAL WATERS
N B Bhosle K Venkat and A B Wagh
EFFECT OF TRIBUTYLTIN OXIDE (TBTO) ON THE OVARIAN MATURATION OF THE PRAWN CARIDINA WEBERI
R Nagabhushanam B Indira R Sarojini and Mary Frances Thompson
EFFECT OF BIOACTIVE MATERIALS FROM SOFT CORALS ON BACTERIA ASSOCIATED WITH COMMON FOULER
Vitalina Mary Avelin Mary R Sarojini and R Nagabhushanam
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
2
This paper desr1blS the use of ti1c~lctrooes to measure pHbull bull _ bullbull ~- l bull bull
iJQ2 chmgtS ~~ 1II1C~olones ~middotilIlg~ ~f~~s i~rsed ll ~ter ~ wlJlch t1)U11ncjotgaI11sllls are grolog SOllie p~tnt1al 11P1~~tions C1f~se cIIatlges to corrosloO at the
lt I )
surlP alllO br1lJfl)dl~ - 1 - ( _ ~ -
ItIiterWs 8nd ~ ~ ~
() poundI~~lt - -
Ttl lec~_ 1n theu bpelsnts hIM bnndeScdbed In
~~~I )iel~~bull~~i~) but n 11 ~ haJe
- Aj ~-
(1) pHel~
ThapHalec~ lII8rII basadQIJ tne-iWof It1tQ (1__)
wre the pHlt1V8recenton of~~l~ 1a fiI)OS8d bull 1Ihart po1nt extlndlngfro- tt lnsulatlnQ (Flljj1) The
pH andmiddot alUlllno~nlcate glau 1cropipettesued lnthe - - ~
electrode constnction went ~ using bull wrt1cal alectrode
puller assntielly bull vart~c81 version of the equiplllltnt
orlQlnally developed by Al8Xllllder and NasWlaquo1953) The ~ ~ -
electrolyte u-d1llllS 1N tel and filllnQ IIIIIS 8aoIpllshad using bull ~ - or
aydnge 8nd a vary fine draIIIn out Qlass capillary
Once f1lled the elfCttocJes were soaked for severalday~ in 1111
tel to allow coqlletehYdroi~s ofthe pH glass lin lnternd refelflf1(e of sllver lrecoated wi til sliver chlorld~~~s
ccinmctedmiddot to a gold plated pin iealed into the imd of the
electrode lII1th~ill~oo rubber COMPleted ei~trodei er~ stated Up downIIIards lna dilute solution of ~r~iIcrcj In allmiddot uper~ middot-temal reference Qf bull~~l
allCtradilllllS UIIIId DurlnQ -~ts -n bulllact~damiddot was c~cted to ind~ ~~F~Y(_ itrgtbull The cStcu1t layautforbotitthiiptl lIlIdpDz electtaifes
~
1a IIhIWi Sn Figure 2 bull
~ -~ - ~ -- - - - I -
CaltbnUariof all alacbodn id Clirr1ed middotOutmiddotbllfGn ftc AIIICPUIIIri inmiddot c- elctmdeiimiddotmiddotmiddot_middot~
ckalng en apars-nt by in nJIalt~ orthey dldriat
pEOCia ~lbblIt ~lntal callbraUon In ~
CIIIIi8S all ~ datafl(WtMI middotmiddotdl
(11) 0ItygIn a1acUocfes bull
TIUI catheter typa elachode use a fiat ended and of duw 1 to 15 _ It conslatad ofa ~ uia P~tJft- wlra
aeelad 1n 9l~ and IIIDadded in 1IPaKY~1o 1I1th a aUler wue KUng a raferance (see FiQ3) Ihen the epOxy hId~
h I of h l~middot careMJy ~ flat to UpQM
bath the plau _the ~lwr 1IIIIIbnre WIis ttn8llPlled
ecrosa ~ flat lind by rep8atedly d1fIPlng Jt into a aQ1utJDn Of
collodion 10 atheta1cohol A Pola1-1slng voltage of m all __
applied bit eachelectrodl and these re not thariuSed for
axper1lllental purposes until middotthe response I118sstllblallihen
bullp
4
bullit tuffer elut1on a~ a eorustllllt
it i)
~~ ~ ~eted IItld accepted for uSe elliCtlodes
it lt gt ~ ~~J ~J _ bull~ - - -gt bull
IIIIiIr~~Jn~IIOWt1on of 02 KCl D~ygen Ilectrodeser _iliry~ ~~)~ 5middot -1 ~ J~- - f -
it cli~rated aga1pst solution I of known oxygen content (as 1 w f - w ~-l _ bull bull
i IllllllaUred bY n of the l111nkler titration thad APHA 1915) ~Lmiddotmiddotlmiddot ~ ~
it Any el-=trodI not lIlCtIib1ting a ilnaar response rejected ~ ~ ~ ( _ - ~
zelia axyg8n lution obteined by the usa of a buffer
i aturated wUhpyrogallol crubbed nitrogen The 100$
~~~ reed1ng obtained by bubbling a1r thlOUltl the ~ ~~bull ~ ~ bullIII burfal far 12 hours at ~~ture 8M pressure- ~t bull
(b) [)cpars-ttal~
All -rs-tts IIIIIl1I Wried aut in atellperature controlled
bulli it
tank pliad an a teflain NYler ted l~ steel patfon ill insidil a ane _tre CIbI Faraday CIlIIIi(fig 4) Tbe~
prav1decI IIf1th a lJrfQII hinged door at the fiant for eccns and[f as gro~nded to earth to scrbullbullnout radio frequencyi intelfuwce The _tar flowing ttmluitI the experluntali - tri pullPed at a controlled rate fro a temperature
~ controlled reservoir For all experillanta involving algae
i llllA1nat1on provided using daylight fluorescent twes if l1d teel and 1Ial1ns plywood plate approxilaately 20 bull x 15
wire Ixposed in the -rine environmantfar several IIIQIIths to bull
allow a heal) growth of IIIICrofoul1ng organi_ to deVelop The
bullI steel plates had been placed utldally and IIIItre exclu8ively
bullt
~~==1$ and ~klOOrganl_ ~ldirig Itltena ~srar-nt of their
OWl The wooden plates 1IIIIIl8~sn the Jntett1ifalteclJQii
-f llllllre COIIered lIIIith ta 01 tt- alp InWfCld1IIPtth
occa1a1onal bryozaens There IIIIIIl8 IWIo lagII quantitibullbull of
~ retained -gat the algal fu-nt nPiate lIIIIIle alsO cleansed and stalJil1aad_ bllton ~
provided On 12haUrly~ ~dllrUght luar~w1 RddtUOlllll steel plat8s ~~In Alii ~dIxk~ lit
Uwrpaol (lie) IIIhare they IIIIIIl8 co~ by middot~Of~
(xtllus edulis) and the lISCld1M Clona intUn1l us
~1IIIIIl8 roved f the pJaqa NP~1id In cl_
ta tor 5 days at cansqnt ~ toIitDUampe
pH WId pOz ~1ane frc the extuMl t~~
the fouling to the Milstrata llllllre ncarded tt elecUadM
middotheld in a rigid pttllpex ~rr1et attechld to II ~ly
Jointed GIl on the lI1crcIIanipulAtor To nciad the muM the
tnraugh the foulJng tI1IIIIerds the ~ta JI_ta 0( 1 _
could be acCUllitaly controlled by talng the tIM clrlw of
the n1pJlator Iftn each lllc~ the DVIII ~oIdlft9 shy
allowed to _tabIU for ane wte prior to ncordJng the
VGltge~
~
bull a
-lt 1
l~to the_ batnacl~ 1ls~ COC)trol - fmiddotmiddotmiddot
recod4l9sof W pH 8I)d pOi at the SIJIIEdepth outside tOe 5hell middot-middotvmiddotmiddotmiddot_I~ lgt~middot middotmiddot~_middotmiddoti 1gt )
wete made
-flesolts ~
(a) pH IIIldIDz p~lllllS
The _re~ts for Hancl p02proflle tluough both algal -d - ~ gt Comiddotmiddot i -
eniIIIalfou11ng are gi~ in Figs Sto8bull --
TjpicalProtUeis daIiilttuougha t of Entronlorpha are ~1n -bull l J ltl ~~ -
Fig 5 - lIhllsttyp1cal d1innal fluctuations CICCUIJ8d during bull ~i bull bull e bull bull
photosynthetiC- II1d non-phOtinwnthet1c lods (Fig Sa) mad _~ d q _ _ __ _ i bull
decreases inpHlIIIIl1I~ IIIIa) fro-theaul-f1lCe of ttWt bullbull bull ~ bull
dcyenI tqwards the abstratacn tdch it enclOSlild (fig 511)
~fal1s1n pH--nted to as-ach as 2 units Over 9 IIIj
dIpth
figure Sa ahoid the thangea in pH -d Pl2 between 11ve barnacles
with depthln a stlat1c envirornent A steady decrease in ~
occurred as the plata IlUrfe~es toiereappro8ched pHWtially
fell on one profile before rislngbut lOSe steadily with daPth
on the second praflle SlIdlar changes are IIt10wI in fig 6b
with depth into the shells of dead barnacles 80th oxygen -d
pH levels were re~at1l1E1ly steadY ~ similar to the 8IIIblent - ~- lt
envlrOlWOt Until the -actual dead shell IIias entered When a
IIIUked drop in both fJ02 and pH CICQ11Tedbull
~llell in rigs rit JllellllUrellt$IIere maltle ~
iOlfillidual IlUs$els withtiRle (Fig 7dlb) In thll first
experisent elactrodes were placed indicated (fig)
arCUld indivicUllalsln genU flawJog ~ter The
resUlts show bull ~tle fIwrease ~ IID2 eMIS bull aiJc hour 1Ucj
The pH~t of the~ II4ttIn ~~atmiddot~
81 (te UlbientpH being 80) but 1GII8 ateIIfIU ~ the
1rtIa1ent aJphon to a6bull 111 iIadIIr ~~thI aff~ of
flOiW 00 local ~ciIndltJans~ the ~
~tadbut tile flow at~ turned Off tar 25i1aur
Jadin thelllddleofthlt ~The nailts g1R
in FlQIb -PD2 IIMtls mae ~y inflQldng CCnd1tJanamiddotbut
fell -zkadly fter alhoit lao w flaws caadawJnit a
~ fall lit 90 alii Levels rapidly ratond onca t IIIIUtchadDli agaJil No ILICh teed varlauon ~
in pH around the eldlalent 1pbn 1iIIflImI a taIdyJncn4IM -
obsmt8d aspec1elly cleM to thIIen1Ml CondltLni around
indivOJa1 ~ lIIhllst of interest are lMa repl_tative
of field conditions fro around groups or clIIIPS The
IlICpIIr1ants were ~ ~ted Wittdn denSe clUIIPs of ala
fouling a plate These r8SIJlts are given 10 figs tie (1Il1Ud
IlUlIS8ls) -d 8b (within byssal thrtiIds) In tatic ccinditJans
amprCUld the _15 u-a1(8$ ~ fellrapldly (45l) reelJ pft
rose sl1rjltly and then feU aftar 2 h llhen flowing condltiors
were resuraed after J hthe ~ rose to the original middotlevels shy
The pH rose lArlcedly at the eltictrodenear to the xhalent
bull bull bull bull bull
bull11
bullbullbullbull
it
I
8
s~ (3) but reriialned stlaadyahd ultilllateiy fell silghtiy (2)
inttie genetal fooling inassbull
~
III1thihthe1JyMli~~ Or~selci~5 IiIfieretilltti($trltus teniis ltO -=IiUlteliter~ ~ ~(Flg 8b) ~ lIIithrrolaw~ cCincentreuoos diCIppea framp1~D lBigtif3S
~bUt toaitnpl~lylt4i 150 IIifIo lIIhfjnficw t8st0ted~middot pH
t wi~1rithi thread ClUIps~lMl1BtO 74 (3) IinIt B~ito
785(2) lri-staUc cinfit1at IoihUst ettn88dgtiofthlt ~
it -flnedf81r1y~at aroOOd17S~ lIIlanf1Dlilwu
rellitoteilpft inltleUyJose (by 04 UI1ts at Ii1ta3) but it theo
bull fall only to startrfs1riq tgi1nattM not theIlXper1ilent~middot
bull Dlseusslont f tMresulblmiddot obta1nIid it ~be cl8lirly sean that f(IIJUngI
bull species vethe abUlty to Gtly IIOdlfy thdr loclmiddotmiddot
bull InIl~ts
bullbull L1t1t end dark period tluctuat1ona in algalts IBre obsarved
end ware lri close wpeeIIIen~ with those reported by Terry end
bull ~
Edyv- (1981) with 1ncnasaln both pH end IID2 llIrJngllttt
traquor1ods lIhilst thlshappened ttM surface of the t pre
bull fU_ts ware frelyprojecting into UJe water -deep intottie
mat nearer to the solid _stratlli whl1lre shading fll1lli light andbull accuIIJlatlonmiddot of deconIposJng ~gallIIBtllrial DCCIINed pH valuesbull fell IIIBrkedly Concurrent wlth this dlQp in pH ilaS II fell inbull
bullbull
9
oxygen Such t~ral and $PiItial va11ations in ~andpft
would certainly qive 11se to corrosion cells middotatmiddot the sUbsttatie
surface ~ly resulting in iorroaien pitting
The r] _tabellc ctivlUee or fau)Jng -=raJrwerttmlbullbull
together with their prapeN1ty to-=ci-1tritla~
tlwlsalves wIll also lIied to 1dcIoenv~ lIOdifktIona
Resplretory activities wU1 tend to ~~ tn-the tft and Input caapanants auch a cinia~nY qatk
1mIertabJatesmiddot ara mlk 1bullbullbull middot the bulk of their nlUagllnoua
wastes ~ excrated as ~ 1heexpftunt ncord~ pDz
end pH cningS amN 1nd1v1Q8l~ ~ ~ri
claady 1nd1cat41 the ~ of _tft~t GIl tt ~ of lnIirollliental changes Inmiddot expr_~ta lllhere no Wlter
--nts occurradthmi a np1d dacreUfi1ll pDz
COIJIdalo be a ~decreae in pH (no a) v fladrlg
condlt1C1ns are r-ed tIwn Ieymiddotmiddotor iIay nat be a 1bIIeIqI1int PH
1nctease dBpwIding ~ the --- ofmiddot iheltr ~ by the
organl1n the cluIIp 0wIgn wui a influenced by the
proxJa1ty of 1nhalent or ~a~ the dlnsity IIId the
tIlJcknes$ of the clullp w111alsa t)IMt wiuence in ~
wetr -N_tbull Hencein thick clampllP aUt and detritus
cCUlllulatlon is are COIIIIiIn ~ theh billa IIIdfbls
restricted Br8IJIIltdow1 gf organtc datrital _ter1als CI3IIIIUIIS
oxygen resulting in recLlcing conii1Uons llleasurlllenta haw
shown that sulphids qulckly build up In such deposits and
sulphate recLlcing bacteria y tie-~ SRBconcentrtlonmiddotin
J -I
II II IJI Ii ill
II
-bull I
11
10
the depOSlts woUld terid to Ile lOW at We start of ~r1ments as
condit1onswere nat favourable (POstgate 1979) The cantlooQUs
prockIctiPn of r1th f~plieUdofeacal_te1alor IlUlpllonill1l
~ts bYpoundilt8~ (~~Iick1ngand 1IkIod1) hOweva ~ t H 1 (
IIIOUld rep1dly altAIr theli11l~t and the rua~ofSRB re
notedtofncreasemiddotttcllll ( 1 SRaIll1taat llaUt 1 x 101 SMIIl-1
aftet bull -th
IUctotJlIIl corrosion canfe=t _ta1sln a1Ullberof Ways
(Iverson1972 fUlllIt end King 1975f 1Uilr 1981) pH can
aff~ the ectivity of the 1IIin organlasinuolved the H 1IIho
hava tin qlu pH range beban 10 12 (Graff 1981)
although tIdS -can be lnf1u8nCed by ather fllltors In atI1tl11n
they lIKIJlre a catbon end nitrogen aurCa 11 various
salts tin alactron accaptor (HedJart ~ idbullbull 1985)bullAil of
U- facton are availabll in the debris -alltting b41tween
ar1na foullng -d the 1lUbstra~ 110 providing bull potantlally
corrosiveanvirorwent
Evan IIIhete such accullIUlatiori does not take place varlable
changas inoxYQ8ll and pH ~trations acrossthe suface of
teel soch as C3I becaustd by foul~ ~ C3I give ti_
to corms1an calla lalildlng to piftlng corrosion
There is an initial tllld8llCY for the pH to rise as a relUlt of
the excretoryactivi1i~ of fouling aniIIal and this 1amp UIIllkely
to give rise to corros1an problems Eventually ~er feacal
h
1
~I
materials and detrltus~ill build up around or~anisms ~re bo~
low pH and p02 devul9ls providing a potentially coJJOampiIu
envinx-nt flaw does dify thh _build UfJ but inmiddotthbull
Ixparlunts carrllldotttid notprlvl(lt it R~~r it
l8Iilated the depth of deposit ~1~~ in t- ~ It ahoutd alao r~~- in f~ population so-1QIJvlWela 111111 be dying or dUd Thlb
~lng~wUl~1Ielacal1Md conoa1ve ~iccandltiorJs-t SR8a 111111 b4I fCllllfC314lng ~y pOIi1desa hlbitt~ HtInd 1tt~ WIi~ul create 1~1Mct Jrcx-ntal dlffarenca ~ ~th l1kel1haOd or pitting
n
References
Albilnder J~T 1 bL~1953~~(9EthII prockIctiPn of IdcroaltJodIIs UIIId in I~ studies Rev Sc1 lnstra ~ 528-531
_r1can Ptbl1c Health laSoc1ation 1975 5taIIdIrd~for the lIIIIIlystS of _tars end taatms 11lth pounddIt1on 1IPHIi-~ 1015 ~taenthSt NW 1IIa-l~~OC 2IJ]J6 USA
Bau-8laquo=k1ng C~IIIG amp 1IIoOd EJF1955 Biologlcal ~ in theestuarlna envi~t 1 EcolClil) of the ~ cycle Kcnlridlkjl Ned Akad -t ~ Ser B~ fi 160-181
cter DB amp SUver IA 1961 In IRafaranctlElectrodl$l EdltolS Iva DlG and l-z Gl lc8dilll1c Pr London Maw Yodlt Chapter 11 ~
Davies PIII I 8r1ric middotF 1942JII1c1oel4iitrodas for-aln local oxygen tension in n11 tissues Rlv Sci InStruII Yo 524-533
EdyileariRJGJ ~ Tetty LA UJ83 The 1nfluenc ~lIlcro algae on corrosion ofstructural steels used 4l~ the 1tIirth
Sea 81odeterioratian sect 336-347
bull bull
~
bull - -- ~It Jf bull
- ~ 13
12
Terry LA I~Ed)VeangtRfiJ 1981 lIi(toal~IOI1CGrmianF~ l)i 1St1oJhemailnEifooiiiigffixed gftshoreIt Bot 1Iar ~ 117-183 lsect~~jlaUoQ1l1 _ ~t Ef)e~gy Offs~r1I Te~lOtjjy Paj)8r
-middotltcntpI)~it$t~l91-16PP~middotmiddotdl ( I
bullbull
iIIalk e1 JL 1971 bull Ion specfie Uquid I~~ e)(charig~ ndcroelactroclels Anal ChIIaI ~8S-Q2 AGraf1~ w~S 9S1~ 1orrosloh InFIntroooctI00 tooffshOle
51~weSoes~~fab~lcat~onIns~allat1on Gulf Wall PI aelUnger EG 1982 The appl1~tlon 9fMl1ikIngmiddotHoust(jfW~Q23g-258middot icroelectrade in the study of pH end PD2 in lIHinli
HerbierC SN fGH~rt~ )RI -Stoctcdalpii Watltif1Sal RS ~1ronIIInb Ub~~ 27l-Z79 1985 Fac~or lontro1l11)9 tha activity ofmiddot sulpMte
bull Wall P 1983 The appllcatlon Md ~ ofiwrJ)e~c~odl shy-taicfii9 tiiCtetili tn~ diiilncj tar infection
techniques for the iiaurnt cif icraeov1ror1liltital5QiCiety ofJebo1- Engi~ SPE 139781-10 _i~~ - ~ ~ i ( _ IKIdif1catlons caued by aGII8 rlnefou~lng -C~ebullbull
PhD lhaaia Ibiverslty or ~tar UKbull bullbull bull -
IIH1nke JA 19fiSa In EXperllRants in Physiology and middotampioClIiIiiilatJift l c E(If(orltGAiKetkutgt ~c-Presa-
Wooltdngton AObullbull Davenport J 1983 pH lind IlIl2 IMla ffl~- j~t~5 -lMIIleath 1IH1na ro-fauUngGqWl J bull Expbull 1
B101 Earlsectsect 113-124 Klnke JAIII 1969b II) Glass IUcroelectJQdes poundx1tors Uwal1~Ui~ OrOllt arid~t Nt JOhn fII11ey YOI1lt thaptar 11
~-f _f~ ~ ~~nt ~ Igt
bull Iverson IIIP 1972 Biolog~ Qrroaian In Advencesin
corrosion scianca and tecIYIology 2EdS FCntaia G~ I bull IIS~et RIIIbull p~ PJaSsLancIoIl 1-42
bull
Jorgensen BB Reusbech NP I Cohen T 19B3 Photosynthesis and strUcture of bIIn~ II1crobWp MC1OIIlectrocki and ~ stud1ea of four cyanabacledalt -~usllWtOl ~bullbull 1~f093
t l~ O~ ~~~ H~ ~9s7~c-gtHatste~iecfn1k van shy pellediftlrtenPt - Stlchelektraden (1-Su
AiJBitiKlificfiliilislflilr)zlIr polrapusdiailJIIissui1g_ liIserstoff(jruQkas fur d1eB lIsUbullbullung der IIliJlttOUf1cillatlon PflftgaHArdI ~fl39
bullbull Mller JOA 1981 tals In EconcinI1c II1croblolooy sect McNbUdbicldatariOtationEd Rosebull A middotAcadelc Praulondcn149-202
t Mller J DAI King RA 1975 8io$~iorat1on of lllltals In lIIi~1al ClflPlCts of the deterloration of tarWbullbull EdsGilbertRJ~ middotloveloCk 0111 bullAcadeic- Presst IILandon 8Jo103
bullt
IPotgate JR 1976 The sulphate-reducingbacteda
Callbridge tIn1versity Press Cliilbrldg8U-K
Reusbecb fUI i Ward DIII 1964 i~~~1Ida5tud1el of
bullt inter$Utilll IIIItez dttm1stry and photosynthetk activity
in ilflaI spring Idcrob1altAppl~EIi~ MerobW tsect4IlU-27lt I
IIbullt bullt
bullbull Ibullt
1
Inmiddot middotmiddot11middotmiddot f~~ a~ -tmiddot bull J~f
I~ W
i5iImiddot J~a t
ilmiddotmiddotmiddot I middotIt Ii
l
~
middotmiddot~~middotlL= g r
Lj Ic
t
i ~
I
gi II cEi ~ I I
1 t rW
D
tR bull
bull t 10
11
r
i
I ~
j
~
I
~bullbullbullbullbullbullbullbullbullbullbull
LV
FigS pH and p02 profiles into macrofoltling eomlllllllitilS dominated by Balm trenatU$ on steel plate 11 two pH profiles Ib) two lIOai profiles leI pH and P02 profiles into dead barllllCle shiella withdetri~Us I I
itIbl r p02 ~ I
nvnHg 11 70
j ifJi eo 1amp0
laquof it 30 s
I 20 N
ca 11 10 a o -- shy i bull
11
80
19
781 o 2 6 8 10 12 0 2 6 amiddot 10 12 dIpdI Qf ~iftto fouling In HighIst numbIII pIatIi
8deg~~
~i c5 iioo5 fiil
pH 8 bull_ ~
~~ ~
_ bullbullbull- pH
-pOz
(cJ
e-Clf ( j~JJlaquo1- --~I ~a ilil - _ I i ]1
~ ri en i ~ oJt ~iJ id tUI
2u1U I shyI t
shy1 2 3 6 6 7 8 9 10 11 12 i shyDepth 9f electrode (mm) into fouling
1 i
1
~----------~-------
~
~-~ lt1~
middotmiddotPmiddot ~
I
V
~
ill
1 Tbellitt~~ of cathodiC protection OO~~lIic~tlO aridrfOflllUlc
ofi AA~Uoullr14l ~1~I
I
ael~lner EG middotIUtcb8l1AJ Dept lt4 lInY1roMe1lal Bl01011
=~~venilt7 IlariCheW 1f139PL $ ~11al1df
r
I
Ablitrict ~~~ e~l billttI ~ted with ~pert1n ant11_1111ampp paint llere itpoii~1n _vatI- In the tl~ld iUIit 1bOioatbrt~ScM Were
1IUbjll~ to qtbocJ~C ~teltt1on byen _ current (~)SJi inci EDAX Iieamp~ta IJdlctecI ~ lachln teoc copper OIl tbePlCtec~ pa1n~ atter14 dQ1a~ protected Pllnt abo ebowed
) reduCed bacteiial activIty butmiddot ~e4 dltoe Iu1d protOzoanmiddot co~~l~tlont~1l IlU l~byenCOCltact not byen dUfwIl00 ~r _baRl lrillolvedln tOXln nil ereio noted
l ~~
1
I
Introduction
Applications of protective coatings and cathodic protect1on(cp~l
are recognised corrosion control methods for IIIBrine steel Itructureso Cathod~c protectlOll preventl conCIsion by Int~lng til8ctrl~l
currentS trem externalllClUfCa to e~~tthlJno~ e~ carrollon feetlans C04U lUChu tUauJ1ng Aelnts a
(
barrIer to ~ the flao of couos1a1 ~aboIHnu~r
Dccurrlng anodes cathodes or wlwit gelvWc CCIIII1 ( 19681
flinger 1980) AnUfauling ~ also JJICOqIOrII~ b1oc~~
Tr~ylt1n nuor~ (~) end o~ Oxide (~) (~ flJ1J
- of u blacld1111~
V _Ut Evans 1981) ~ ~
end growth of bipfoullng 0I1I)IIIII1IIIIe Wdd~ ca ~ of
paint rusendmiddotmanemiddot ~(r~ til )
As corrosian ~t1on ~ bICCIIIII -an ~1tlQd ~ of
antiroullnQs 1 conjunction _ith CP have bllClIOr ~
resultIng in reseudt 1rwestllj111tlng trw COIIIPIitlbUlty or VU~ ~t t _~
rOratlation to dlfrert levI ot CP (~Eelf 18110 ~
~t 1980) RelatIvely 1nIestiljlllUans have ~ urtw ~
or CP an thIt anUraullngpert~ of palnta
In thIt preseot 1nlIestioaUon tV~iln111 ronutlon ud
cOIIIHrclallY incanjlntlon wIth CP was ~t4IC~ Iw I sraed current (-966 l1li) arid exposed at the 1 Strait IInQlbullbullbully
~t1ons weremiddot 00 the IIhcIrt bnifects the PprIUd_
had an a) the various COIIIPQI1eIIts of n pe1ntf1l11 end b) l1oft~
developaentbull
bull bull ~
~ ~ - f
bull ~ _T~~-middot~ ~~- ~r _ -$1
(1) ~rlttionor Painted Steel Billets middot_ ~~tmiddot_tlJ middot~hmiddot~--~fmiddot ~ middot~A~-~~middot - -~ ~
Thirty-al ateel blllets (8-10aD2 end l depth) were cut frollbullt ~middot~middot~middotmiddot~middottrSmiddott ~jS-~ It bull gt~ tfl - middot--imiddotmiddot ~ t1dItrd steal plates The dimenSlonsof -thB butits-lim chosento
bull -~~ middotmiddotgt2h ~ ~-Imiddot ~~rt ~- -middoti~~ ni ~~middot1 l middot-~t~~middot _
(a) fecilltate amiddot Ie detrucUve -thxI of prePraUonpdor to ~~_gt gt~ k7i ~_ t~ 1--1- _ ) - +--_~ -_~bull ming electron ~ 4II1CI (iialloWdirectobs8iVatlCnS of the
- i lt-- ~ gtbull -o1e billet 8Uface
gt r~middotmiddotj ~L(~middot~middot rmiddot tmiddot J ~ - ~ - lt _ bull Single non-standard (1hIula~) nickel lIIireSmiddot Spot Welded e1Cing
bull ~1deJ ~ b~i~ ~j~~ tri afl~1b1ipOi~ f ~1t1 ~4middotmiddot f-gt -~ ~S-- ( -- - - _bull resin EactI billet then Undb~ tI1d prtiladlllltha5O __ eyilr
bull ~~_ ~~imiddot ~1iil h~ intlfcili~ ~eonbt1nlyi Tr1butylt1n F~id(18n)~~~O~dettu20i~blcicld8is 1IIfii bull i - ~ ( then 1IPI)l1ed to glvaadry Ulil tIi1c1ltnOss or 1(1)-150bullbull
tbull (2) IipreSSeCi Current Syat
bull A rigid ~tic due 1I8S drWedllliihwrtv-s41 liiIihOieS IIPProX~
bullbull 05-10 cia episrt ~ Pilnted ampUl8t Wsfbed IntaPQSIUon ampy
threadlng the nickel IIIlr8throu11 the dinto a tarproot junction
box Eliltilen of th8~1l1ii (recelving ~t-d curnritlere IIIlred
Into bull block connectOrrecelvlng dc trOll is potentiOstat The
rainJng iontroi blUets Wet8fbedfl11i1iiy lIIithln thejtlytlonbox
i~ityd~c IkiliPly ~~ ~lat1ruaWiode 8ncIacalOliel
refer~ electtodetoiere fllied ~ the per1aetat of the plut1cdlc IIIId
connected to the pot8ritlostat The desl~ of the potentlosbt ailoiolEld
~PlDteItlO1TClJrnnt to be MIllntained at 8prampset levtl of 966 II1II
c
r~
Grid raf 1J5s-t 114iH 563724 dud 0 Juna
Alolecule andbechr1a in general are ~ to
sutface lI1eroleyer (nlUSton) of _ter (~bullbull 191KJ)~To avold
contaJnatlonof ttw painted surf by U- -vents
blllets Were s-nact in a 1 conte1ner or atc1le (02i ) f~~nd
ter Iih1ch then f9ICI in tv 1IIhere the b~
be r-oved undeNeter El~~ bllleta fa Mcb trtai bullbullel
removed over a 1 dtq (336 hr~ expoeure perioda
each being ~~bn uposurnof 1 5 11 iA 168and DlIIauz1r
Arter field axpauHli the billets wan
fUtered _tar to ~ 1_ debtlflMd far 10 aJnuta ~
wv ~~~endattid sdbUiUd(otO
~ ~l f~ nn1ng el~~ --
lOOted onto 1middotCIi ~bullbullWIsend carbonmiddotcoa~nmiddotmiddot~middot f
each billet Ws~ Iis1ng a ccr1dge ~el~~
(5360) IIIlth anergy dJ8perslve X-ray alcroenelre1a (mx)
_lysis the les Wen reNed end coated III1th vold to al]Ql En
detailed sat of the alaquofa1l
(4) Lfiborattux ~
Painted taa~ bUletalllllre pnparad ~
in a Mrlne reclrculet1ng flOlll _ ~
f1Oll the ~ ~a1t~y Tha~~Un bullybullbullbull
10C M1nta1ned ~~t1as of ~t~la protazOllbull~bull10shy
(Entermap) ~ ~~~middotNlt~Eamp1~ __~and ~ r - ~ -
certaJn ~lc 1IQ1~ 1(1 the
the painted
~
thre repl1catft fn
plKed 1nto(022_)
- thIine1J~~ -lt bull
Atter mAX
(1-2) and i-n-t bn collected
14OC
Pain~
-
12 daY~ exposure and prepared as in d)~ - t
~I ~~~-- i~yen 1~ v
Rflltsi ~ _ ~f-~lI ft j bull _
rt~ll0~~~llY~~~~peln~stetsl~11i~ex~sed for
y~t1~J~ ~~~~1~1~~1UII~ya~kooJ~~~lt ~ ~~_~~I1J~ ~~~tomAX -lysis ~era than gold ~ bull bullbullbull c - _ bull lt
~~fllr ~~~ vllUll ~t1ans of the rfee htures and gt Co
~f~ - ~~~ ~f ~ ~aobta1Mdue IjIll811 iI1Flg 1~2 The IIIIdn ~~ ~ lt)ji9 )tmiddotmiddot - ht
- bull
~I rIIplt~ tIt8 -~ cOlllponanta of the pei1t 1bullbull WelIIlItr1x $ilj~t-~~middot - ~ _ -_
-IUch PVCbQed end rap~13y the chlorlna peak the
i~~~-~~-~V~~~-t18 ~~~~ ~~ Th8
ral~~i~ JlrQP(lrUo1sof ch 1Il-ntal paek~ll1d l~t_ Q~7 -gt- ~ _~ -- gt ) gt -_ -lt ~~ the 0 -end _ xprtsntabullTIwsa changes are -r1sed ~t~ lt~ _ ~ 1nT~ IThe8llia$lb npresentthe chengelI 1n~t1V8 p~lons
ti~~~ [ ~gt 3 - - - _ - _ -
Qf nellllllllt within We surface layars of the pe1ntmiddot ru The
~f~~thtJn -d ~rfeli avr the 336 h11cxI ampScanbe
SII4iIl r~ fig 3 al~ tht rta of~ dirfers tJetwaeo thaa ltmiddot~1- 1ft~- ~ _
IJOl appfiai-ed to ~1nlt1elly until ftar day 168 it fall
~~l~ 1h8relllllrelgn1fi~ diff~be~ qrOtactlid and
proieCted (at -966 ) piIIllnts tor lioo COPPer but not folimiddottin
Ibi~tlons Of the Pain -urface 8hoWad _Pil ~ witilt1M
(Plete I to IV)~ 1hePllat1na (~) pelnt(Phta x and b)
Clearl~st-i~~r alots on the SurfllC8 IiII1kh~ter1sed ~- f hlt1 lriitUl tin conCantrat1on~ lllariy slotsre Partially
~OIIe~ ~ tile _tr~tedal middot1lere rs ~an~ densely packed
grerltr~ ~th 1ar~(iIa1niy cotifIIr) 8ntt -ampniBil (lIB1rily 1rori)being
middot vl~l~~~~~ Dtt~i~~~t~ r~ ~f~~ I~t seawatllr for 11 n IIIOre slots appeared and th~ ~~ess _trb cwer
--if -- V~ c~~ f(i ~~~i~
(Plate Ie to It)~ the granules dld not protrude from the~rtace in ~ ~~~~j1~r~-ffJmiddot0~~
suhap~~r Both 0 nd --960 at ~~pa1nts ~ - fr ~~~~-w1-U~ ~~t
sl~llrat thi stag~ (Plates Ic-d II1d xrnspac~IlYh middotmiddotWith middot middotaJ~cmiddot gt incraallO ~alolre t~ difrrqcbullbull middotbatIiI8IImiddot the protected fjd
bull ~ ~ ~t~
IIllrotacbldpaint ~~lIfIt~ftitr~ h alQt~ p~t bull u bullbullbull J ~~i~~ 1~~
lIIith IIIuper lIdguon u 4lrotatad piIJnt (Pl8ti1 II) 1hD1II8S
Ill )1IMdw1lillS gnnJla~on f-r~~ dpert1cl ~ - - - ~~r ~t
Onthe 966 1IIpetnt t-var tIIO typeaofan with diffannt c t
~hIId~(Plate UI a to c) One hIId tIII-1 bull bullbull e _ bull
fUll wJ1hleas abvlaus alats the o1fwl bed C18UU1IlOt ~litUi middotC c ~ ~
less ~ t11 h Gnnulatlan __ but ~lIiIZly def1nlld in
middot~ latter Afte fila bed tiIIrtIId 10 ~are ~~j(
extansll(tlly ~ the~tIId(O at) tIIIIple lIIIdngthe~ - ~
middot orarallet1ai leas claaI (Plate I~) f u II1ao CQ1tJAJ11d to ~Gn - -
the -966 at 1 (Plata III d tor but 1JI8V8n 1nittNdcneabull -
Althatqa -middotslots wrs abvlaus becaI1ng 1 cwampr aiI- the orwWat1an8
f1nailyftar 14 day exposure ~ fUll drMIlapIIIent on all la - ~
are extensive with the 0 lhawlng both alota pore ~
OVIKgrOloln (Plata II c to f) Gnrulatlon 1bl to ~ gt~t~~ i
or the th1cknats of tho ma ~the ~ at ~ (~~~~ themiddot - - _ middott ~ ~ ~ - -1 1 t
middot f11bull IIIIIS IIOre extans1 thIIIn at dIlya but IIIIIIIY bull1Q~ tI st11l ~w -middotr~ 1gt ~1 ~
obvious fIIany ~ loI8re still p~ yen~~~~~~~r
than before Poiu appearedto be f~lf~ila~~c-~~~a (Plate IVf~ label 1) IIIIl1ch broke~i(2)~ ~~~~
t t
bull ~lS (3) ~ rflC~~ ~i~t~~l~~KI~ t~ldiIiiicyul~loIiih ~~Rht ~ )~- -~ - ~middotd 40 ( gt ~gt ~d lt1 t ~ Jt ~remiddot4amp tha paint trix cavity bottom eroded away reVe81~ng ore
t ~~t~ por~~ ~l~ P~]Y p~~8dby di~lutiaA~ ~~~~ti~Jr~gt~ta(l~~Siinll1cl1Ud~f iUiIiI reriatlGllbullt
bull a~~ ba~tid colCriAtloriccic~ GIl ~~~d ri -966 - ib~~~amp catQn1iti~~t~I~~(~t an ( ~
t the ~~~palnts (PIa_taiiI d~~h ~wh ~ n td have ~ CamplluJMitiIVc) ~~middottti o~lgtnalrf tlt of hIght
~ l=~A ~J~rW tYtles~ p~fiiilriQcOCCold to~Ja to jU-t ~jmiddoti~ai dlatClilS ~ir -jlati ~tbullbullent aNit ~ t ~~ th8middotf~ld~ies~~t~on~ ~ -966 prQ~~~(P14ta Iii~)Omiddotiliof~m1hiiHAiiipieat ere~ll-(piitir~ p-iot~iilans(PiA-iII~ end c) nbullbulltiledt dl~by1i~~ end wirinali1sheii ~cMI~bY thI tili
bull In t-~~ ~1i1nt 1oIIIeHfd~ ~m1IS (15OClind a t ri~r-dlatoa ~lor P~t dlataiOlonlt1anGIl ttw-966
t
bull ~
pr~t~ ~is~ih iion obitJauIJ A ~rot -=1 ~ ~ i __
reprasenttild on the 966 painted 1iUrfc1ncluicunljj ~ rwt
Achnanths Colonisation by bacteria 1n the laboretory bsed
bull eliperlMnt tollMct sbiUar pattern to the fild ~
bull bull Dilicueii
bull bull DUt 1Iidnt ~lona Mve dlttllrint ~~ ictian
It ~ JntM l1tenture ttt Iff yen __ JIll the
bull ~ bicins ani 111 ~t INchlnil (PhillIPs 1t73t EWene~ 1981
In ci1ntlHt tJWY ~tit ~Wl1c iIuCh as 1rbiltyl tirI
~ lite wAlII]ltiClr dlsP8tfu~ middotthiirsttilll~intaJn~ a
bull ~
lIIifoflll ampalCentraUon through the filII 10 thoIt biocide rel_ at tha
paint surfce 1fI$Jlts in IIIOre ditfusing ~s to repla~lj -iflJ~it
resulting 111 bullbulltNdy lr leechlng rata M1llat this IIIIY be middottruIi ot
tin ~ it was nOt true ~ ttw middotlBlf fOZWJlatiGn ua ~ bull - i - bull 1(1
ObSfvt1onundr SE clarly ~ tM t1l1 to be present
~ly lIS OIIIdle-llk parUc1~~t tn lUlfiice (Ito
IV) bUt usa ~ the dIpth attt fU-middot ~y~
If one considra the concn~rtlonmiddot chMge 111 1n ~111c COKICIl8rlts fro- the paint tU- i bullbullbull tMWei b~ ~ Si6t arid
tI-~t iron oxidt 1~ can bt (Fl_ 3 IrId TIibl8I) ~t ~ ase cona1derllble d1fftrenC8s ~ a Un decz bcdYMr ttw p1iod IIIIwraS lron~- an unt Ught inCr h ~ incqJper ~tlGn occUrnd _idly at ttw ~~ Gfb ~ period ~ out ttwn fu nPldly ampQII1n ~ umiddotirid II phast This IIIOUld bt tIlq)8Ctad It cantact l-=h1ng we occun1ng
IIltfIOS8d pertlcl of copper would lMch rapldly on t-n1on 1n - shy JIsewtr There would thlll1 ba bull J-a a1iltUy leu avalUba
i1 ~ ~
_tedal becfM uposed Thi tIOUld ttwn IIllCh Thla pulSing efttct
IIIOUld be expctad to -oth OIItlllith U- putic1 __ -lllible II tot 1~ GIl a on r-x _lao Thla teet 01 put1c1e 1tUng
can tit GIl the lIUrlace ClIlC8 it ~ a pan 1amp t
a 11aM bet in Plat I to IV IIaIrIg _ lit la_ II ~ 0- ~s _- PIate IW $llldl ~bull nshyot poRI flllNltttn Al~ tin ~ lou rete thtn I bullbullbull atill Ivwnce of bull al~t pulsed 1Itue rrlict tII1IIWd
elqlCtaG inIl ttwauUr ~ I I I
0
~ i
~ ~
~I J
bullbullt bullbullbullbullt bull bull ~
bull bull
I
bull t
~~
The same seqiHllle of cevity follliltloii (intttecaseofthetestjjltle
slot- shaped) fClliOileciby SecOlld8rYtlolesfotlllingin thll cavity
bottollis releasing further deep8rtox1ils (Plate XV d and g) As
cavities_re ProdICed ~ tiecaaIelnvadedby_ter~ onelilauld
expect 1ncreaa 1n1IOdha and dIladne lMls t the t1met
which the toxlM uw ~ Ths fCMld tCtHi eo (Flvmiddot 3)
~rttno the vi that_tat ~t1on otthe~ ~lY
il1Cl1l8S8S Idth UIiJ
Differences f~in toxin li~~to Ini 966 ill 8811P~ F ~ bOth Un end ~ lcMIr levels afillW
IIIIIlfI ptaInt(~1~~1ntha~~ ~~f~~ cst rklld with tbiIccpper ElJIc~l ta~Uona invalvinv
w pl8cId at a~ht~~middot3lO~lth Un cqJpel bull
(-960)10 ttW ~af this expei1tIIri1ts thIsiorelfJltdt -
that cqlp8r b8IiIId ~ IIiIOUldProc-ln f41ip1dly t thellllgllitiv
patentW _lied () as 1t 11 further f~1t ~ witt Un
III11ch is u-tat aqulllbd Idth
it If ttda 18 eo~ ~
~ )
frOll1 paintscould bl anqnced by an illprd current clithodic
protecUansysteipos1bly ahart_nino te 11fe of the paInts
antifouling ability Bacterial coloni Uon IIIU rtdUCed on the -s66 paint This
could have been due to unflSllOUlllble electrostatic cqrge on th surtece Th1a effect CIIl pley en IIIportant role in the t~of
bacteria to alrraces (lIIarsMll19E1O) As bacteria appear to IhiJw
greater toxicltyto copper (DMpsey 1981) it is Klre likely that
enhanced cqJper leacll1nl On the -966 AIIIPl c8used the recluCed
activity The IDlol8r toxicity ofUn to bacteria (as reported by
~ -
DelIIPsey Hle1gt or its lowes leaching rat was sUpPorted bythl
relatively rapid recolonisation of the slots by lIIicroorganiSlllS ~ftar thE inlt1al aurge of leachi~ Thailllpressecl current dto ~~ the oppoSite _ff~middot ~r on dtta colonstJon l1lI-1- in the
JIbcJrtory~ta larger rUDIIra f~ fit pro~td ~ther than uriprotected Pafnta
Ccnlus1one
1 tiild-a 18 presaIIUd ~ thattJn~ ~~
~llNChng notd1rf~ ~
2 TIleuseofCP~ ~CUIftnt(at) on1h1a ~fca41ng ~- ~
pa1I]t~ to ~_lAIIach1ngn_ at toxiN - -
3 leIJstaPidbectar1al ca1on1aatJon acaampmId an ~bId Pt1ntJ ~
4 JItore larger alcrofauJing (Oiat_ and PzOtoaoana) Gr~_
occurred on the protcbld~
I l
I
bull bull ~
mfEflpoundaS middot~-t9i _jgt- ~ _ _ ~sev M~J~ (1981) (cilonJzatiooof alUf~lng lnts by _rine
Jbull ~teda9g~1ta IIarina XXlV~ 185-191
- - gt 0 Ie 1 _ - _
poundV8I$i I~lf (1~P~h 1II8rin~Ugae and Foul lng AReviw ith
t p~l-i9Jrlr ~iiMdilto5hiP-fQUllfCiH 8ot~ ~r~ ~~~ XXIVbull lIPwl~J11A
Fotteth CN~R ~kk8nGB IWPIR1seJCiiiroaic1lihd ~e ~JIIth CII Offshore St1ucturu ElllS ~tjI~f~1Dt19i4
- - ~
UKC (1976) I~races ill ~W Ecology KIuverd Univbullbull middotraabullf~iddge ~bull C~~jII illt)Ia(t RC (1980) totinllsa Cattod1CPrOtect1cinbull middotmiddot8AtEmiddot~6haOCIlfif~ T~ p No 108 ~ 18 ~ 1IIHf bull
t bull lt ~J~~~ __ ~
Norkr4irulB(198D) Advanculn UcrOfj~lpound~ltiOyVol 4 (AlItxlaquoIder fl Ed) PlerIw Press lcxIdon~ lIP~ 51-85 ~ l~l - bull bull
PJijllt~i At(fm)~mr~~JnI~1fqul1nQ JlalntlJ Riumh ~JnOrpnic toatJngs 2 192middot
)~J gt0 c ~lt -- ~ __ RogerS~ Ut (1968) ina ~(lnt~fbItliP4ili GI COnoslon ~a Technology)bull bull
Sl~iorr~ VP~Ranald~ Rctd~~ (1S80)_ ~ Experllattal Stud1aiRelate middotEffect or Cathod1c Protict1Oi withmiddot terta1nr1c
Coating Syst~ 12th Annualofr T8Id1na1ogy Co1fItHnC8 liauston TIIlt p 9-12
j
fIG1 sPct FromMon IT8TJCuZOI Int Lliu EOAX
FlG~ Fro TlTCu20Ar-4 Dy Field Ellposult I~ir
il
I c
R hi -00amp=7
-I I I I I I I I I f
iii
bull bull
bullbullbull
I J
J
I Figur 3 Thl chan In reltl rqpoIQnlof -ariOtimiddotjOmponents of the paint film with time Thl vtic~1 ampkiS unl till _n ofmiddotmt Illment per unit OJ fe 50all Cl Na
Cu StI Feeshy
PlistinllFe S7~J6 704 3~74 908
f
r e ~ ee~
middotie ~lt - ~ shybullt bull ~
t -~ l Cu~ o ~ f
t --- 3igt - bull 0 J Sn
cc
t ~
~
bullo
~ bull e
bullbullbullbull bull ~
bull ebull a QI 1- bull 0 40 431282 316
101 dmllIII)bullt bullbullbullt
hr 0 ~~bulla 5~16 ~~ -~~ i166 4S i83 8~1B
24tu 6~480 SD~83
111middot 1~1 OS6
~ 5114 6~69 -51 UIamp middot016- lt ~rmiddot
1d o Mea 818 21 021
~Z21 199 1~1Qmiddot ~Jgt2 058
14 d o ~~ 804 Q~B8~~~a~~t~middot 685
866 372amp 8amp4 OS _i~11[ ~~64
Tttile 1 A81UvlL pzQJJortlcw 0 vxl~oI ~~bull Jn 1I1Dtitctel lt-1166) end lqIlDteCt6 (O)middot~af~_~middot_lodEDf~ middotmiddotmiddotmiddotmiddotmiddotmiddotmiddotmiddotc
~ ~
_----I~
J
I bull
I
J bull tmiddot
7o CONQRESOIMTERNACIONAL
I D~CORRO$IOr(IAAINAEmiddot~USTACIONES~v c)
t 7thflfERHATlONAL CONGRESS
ON IARINE CORROSION AND fO(JUNG
~CONGRE$INtERNATIONAL DE~CORR~ON ~E ET QES~SAUSSURE$
i t
~ ~
bullt
t t
UNIVERSIDAD PourtcNlCAVollncio7J1 NiwiemIif~ 1988
(SPAN
SECcl6lfn
BiOIogro marino Morine biologY ~1i1orine
Theunpub1ishedJllBIlu8criptEl have been reprod~d asr~ielved from the
authors The SCientiUc COlIIIBittee take not respoi~ulbll~~Y for any shy
error or omssi9n
Laa ponenciasno han side publicadas y estanreproducidas tal y como
sellan recibido demiddot BUSmiddot autorea pound1 Conte Cientfico middotno se responsemiddot
bil1za de cualquier error ~ om1si~n
bullbull _ bullbull_ bullbull ___ bull $_ _ L _ - _ --- -__
SESSION II Marine biology
Friday 11th November
SURFACE PROPERTIES ON NONmiddotTOXIC ANTIFOULING PAINT FILM
K Nanishi M Murase Y Yonehara M Kishihara and T Hirama
MICROFOULING ON ALUMINIUM PANELS PLACED IN THE TROPICAL WATERS
N B Bhosle K Venkat and A B Wagh
EFFECT OF TRIBUTYLTIN OXIDE (TBTO) ON THE OVARIAN MATURATION OF THE PRAWN CARIDINA WEBERI
R Nagabhushanam B Indira R Sarojini and Mary Frances Thompson
EFFECT OF BIOACTIVE MATERIALS FROM SOFT CORALS ON BACTERIA ASSOCIATED WITH COMMON FOULER
Vitalina Mary Avelin Mary R Sarojini and R Nagabhushanam
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
bullp
4
bullit tuffer elut1on a~ a eorustllllt
it i)
~~ ~ ~eted IItld accepted for uSe elliCtlodes
it lt gt ~ ~~J ~J _ bull~ - - -gt bull
IIIIiIr~~Jn~IIOWt1on of 02 KCl D~ygen Ilectrodeser _iliry~ ~~)~ 5middot -1 ~ J~- - f -
it cli~rated aga1pst solution I of known oxygen content (as 1 w f - w ~-l _ bull bull
i IllllllaUred bY n of the l111nkler titration thad APHA 1915) ~Lmiddotmiddotlmiddot ~ ~
it Any el-=trodI not lIlCtIib1ting a ilnaar response rejected ~ ~ ~ ( _ - ~
zelia axyg8n lution obteined by the usa of a buffer
i aturated wUhpyrogallol crubbed nitrogen The 100$
~~~ reed1ng obtained by bubbling a1r thlOUltl the ~ ~~bull ~ ~ bullIII burfal far 12 hours at ~~ture 8M pressure- ~t bull
(b) [)cpars-ttal~
All -rs-tts IIIIIl1I Wried aut in atellperature controlled
bulli it
tank pliad an a teflain NYler ted l~ steel patfon ill insidil a ane _tre CIbI Faraday CIlIIIi(fig 4) Tbe~
prav1decI IIf1th a lJrfQII hinged door at the fiant for eccns and[f as gro~nded to earth to scrbullbullnout radio frequencyi intelfuwce The _tar flowing ttmluitI the experluntali - tri pullPed at a controlled rate fro a temperature
~ controlled reservoir For all experillanta involving algae
i llllA1nat1on provided using daylight fluorescent twes if l1d teel and 1Ial1ns plywood plate approxilaately 20 bull x 15
wire Ixposed in the -rine environmantfar several IIIQIIths to bull
allow a heal) growth of IIIICrofoul1ng organi_ to deVelop The
bullI steel plates had been placed utldally and IIIItre exclu8ively
bullt
~~==1$ and ~klOOrganl_ ~ldirig Itltena ~srar-nt of their
OWl The wooden plates 1IIIIIl8~sn the Jntett1ifalteclJQii
-f llllllre COIIered lIIIith ta 01 tt- alp InWfCld1IIPtth
occa1a1onal bryozaens There IIIIIIl8 IWIo lagII quantitibullbull of
~ retained -gat the algal fu-nt nPiate lIIIIIle alsO cleansed and stalJil1aad_ bllton ~
provided On 12haUrly~ ~dllrUght luar~w1 RddtUOlllll steel plat8s ~~In Alii ~dIxk~ lit
Uwrpaol (lie) IIIhare they IIIIIIl8 co~ by middot~Of~
(xtllus edulis) and the lISCld1M Clona intUn1l us
~1IIIIIl8 roved f the pJaqa NP~1id In cl_
ta tor 5 days at cansqnt ~ toIitDUampe
pH WId pOz ~1ane frc the extuMl t~~
the fouling to the Milstrata llllllre ncarded tt elecUadM
middotheld in a rigid pttllpex ~rr1et attechld to II ~ly
Jointed GIl on the lI1crcIIanipulAtor To nciad the muM the
tnraugh the foulJng tI1IIIIerds the ~ta JI_ta 0( 1 _
could be acCUllitaly controlled by talng the tIM clrlw of
the n1pJlator Iftn each lllc~ the DVIII ~oIdlft9 shy
allowed to _tabIU for ane wte prior to ncordJng the
VGltge~
~
bull a
-lt 1
l~to the_ batnacl~ 1ls~ COC)trol - fmiddotmiddotmiddot
recod4l9sof W pH 8I)d pOi at the SIJIIEdepth outside tOe 5hell middot-middotvmiddotmiddotmiddot_I~ lgt~middot middotmiddot~_middotmiddoti 1gt )
wete made
-flesolts ~
(a) pH IIIldIDz p~lllllS
The _re~ts for Hancl p02proflle tluough both algal -d - ~ gt Comiddotmiddot i -
eniIIIalfou11ng are gi~ in Figs Sto8bull --
TjpicalProtUeis daIiilttuougha t of Entronlorpha are ~1n -bull l J ltl ~~ -
Fig 5 - lIhllsttyp1cal d1innal fluctuations CICCUIJ8d during bull ~i bull bull e bull bull
photosynthetiC- II1d non-phOtinwnthet1c lods (Fig Sa) mad _~ d q _ _ __ _ i bull
decreases inpHlIIIIl1I~ IIIIa) fro-theaul-f1lCe of ttWt bullbull bull ~ bull
dcyenI tqwards the abstratacn tdch it enclOSlild (fig 511)
~fal1s1n pH--nted to as-ach as 2 units Over 9 IIIj
dIpth
figure Sa ahoid the thangea in pH -d Pl2 between 11ve barnacles
with depthln a stlat1c envirornent A steady decrease in ~
occurred as the plata IlUrfe~es toiereappro8ched pHWtially
fell on one profile before rislngbut lOSe steadily with daPth
on the second praflle SlIdlar changes are IIt10wI in fig 6b
with depth into the shells of dead barnacles 80th oxygen -d
pH levels were re~at1l1E1ly steadY ~ similar to the 8IIIblent - ~- lt
envlrOlWOt Until the -actual dead shell IIias entered When a
IIIUked drop in both fJ02 and pH CICQ11Tedbull
~llell in rigs rit JllellllUrellt$IIere maltle ~
iOlfillidual IlUs$els withtiRle (Fig 7dlb) In thll first
experisent elactrodes were placed indicated (fig)
arCUld indivicUllalsln genU flawJog ~ter The
resUlts show bull ~tle fIwrease ~ IID2 eMIS bull aiJc hour 1Ucj
The pH~t of the~ II4ttIn ~~atmiddot~
81 (te UlbientpH being 80) but 1GII8 ateIIfIU ~ the
1rtIa1ent aJphon to a6bull 111 iIadIIr ~~thI aff~ of
flOiW 00 local ~ciIndltJans~ the ~
~tadbut tile flow at~ turned Off tar 25i1aur
Jadin thelllddleofthlt ~The nailts g1R
in FlQIb -PD2 IIMtls mae ~y inflQldng CCnd1tJanamiddotbut
fell -zkadly fter alhoit lao w flaws caadawJnit a
~ fall lit 90 alii Levels rapidly ratond onca t IIIIUtchadDli agaJil No ILICh teed varlauon ~
in pH around the eldlalent 1pbn 1iIIflImI a taIdyJncn4IM -
obsmt8d aspec1elly cleM to thIIen1Ml CondltLni around
indivOJa1 ~ lIIhllst of interest are lMa repl_tative
of field conditions fro around groups or clIIIPS The
IlICpIIr1ants were ~ ~ted Wittdn denSe clUIIPs of ala
fouling a plate These r8SIJlts are given 10 figs tie (1Il1Ud
IlUlIS8ls) -d 8b (within byssal thrtiIds) In tatic ccinditJans
amprCUld the _15 u-a1(8$ ~ fellrapldly (45l) reelJ pft
rose sl1rjltly and then feU aftar 2 h llhen flowing condltiors
were resuraed after J hthe ~ rose to the original middotlevels shy
The pH rose lArlcedly at the eltictrodenear to the xhalent
bull bull bull bull bull
bull11
bullbullbullbull
it
I
8
s~ (3) but reriialned stlaadyahd ultilllateiy fell silghtiy (2)
inttie genetal fooling inassbull
~
III1thihthe1JyMli~~ Or~selci~5 IiIfieretilltti($trltus teniis ltO -=IiUlteliter~ ~ ~(Flg 8b) ~ lIIithrrolaw~ cCincentreuoos diCIppea framp1~D lBigtif3S
~bUt toaitnpl~lylt4i 150 IIifIo lIIhfjnficw t8st0ted~middot pH
t wi~1rithi thread ClUIps~lMl1BtO 74 (3) IinIt B~ito
785(2) lri-staUc cinfit1at IoihUst ettn88dgtiofthlt ~
it -flnedf81r1y~at aroOOd17S~ lIIlanf1Dlilwu
rellitoteilpft inltleUyJose (by 04 UI1ts at Ii1ta3) but it theo
bull fall only to startrfs1riq tgi1nattM not theIlXper1ilent~middot
bull Dlseusslont f tMresulblmiddot obta1nIid it ~be cl8lirly sean that f(IIJUngI
bull species vethe abUlty to Gtly IIOdlfy thdr loclmiddotmiddot
bull InIl~ts
bullbull L1t1t end dark period tluctuat1ona in algalts IBre obsarved
end ware lri close wpeeIIIen~ with those reported by Terry end
bull ~
Edyv- (1981) with 1ncnasaln both pH end IID2 llIrJngllttt
traquor1ods lIhilst thlshappened ttM surface of the t pre
bull fU_ts ware frelyprojecting into UJe water -deep intottie
mat nearer to the solid _stratlli whl1lre shading fll1lli light andbull accuIIJlatlonmiddot of deconIposJng ~gallIIBtllrial DCCIINed pH valuesbull fell IIIBrkedly Concurrent wlth this dlQp in pH ilaS II fell inbull
bullbull
9
oxygen Such t~ral and $PiItial va11ations in ~andpft
would certainly qive 11se to corrosion cells middotatmiddot the sUbsttatie
surface ~ly resulting in iorroaien pitting
The r] _tabellc ctivlUee or fau)Jng -=raJrwerttmlbullbull
together with their prapeN1ty to-=ci-1tritla~
tlwlsalves wIll also lIied to 1dcIoenv~ lIOdifktIona
Resplretory activities wU1 tend to ~~ tn-the tft and Input caapanants auch a cinia~nY qatk
1mIertabJatesmiddot ara mlk 1bullbullbull middot the bulk of their nlUagllnoua
wastes ~ excrated as ~ 1heexpftunt ncord~ pDz
end pH cningS amN 1nd1v1Q8l~ ~ ~ri
claady 1nd1cat41 the ~ of _tft~t GIl tt ~ of lnIirollliental changes Inmiddot expr_~ta lllhere no Wlter
--nts occurradthmi a np1d dacreUfi1ll pDz
COIJIdalo be a ~decreae in pH (no a) v fladrlg
condlt1C1ns are r-ed tIwn Ieymiddotmiddotor iIay nat be a 1bIIeIqI1int PH
1nctease dBpwIding ~ the --- ofmiddot iheltr ~ by the
organl1n the cluIIp 0wIgn wui a influenced by the
proxJa1ty of 1nhalent or ~a~ the dlnsity IIId the
tIlJcknes$ of the clullp w111alsa t)IMt wiuence in ~
wetr -N_tbull Hencein thick clampllP aUt and detritus
cCUlllulatlon is are COIIIIiIn ~ theh billa IIIdfbls
restricted Br8IJIIltdow1 gf organtc datrital _ter1als CI3IIIIUIIS
oxygen resulting in recLlcing conii1Uons llleasurlllenta haw
shown that sulphids qulckly build up In such deposits and
sulphate recLlcing bacteria y tie-~ SRBconcentrtlonmiddotin
J -I
II II IJI Ii ill
II
-bull I
11
10
the depOSlts woUld terid to Ile lOW at We start of ~r1ments as
condit1onswere nat favourable (POstgate 1979) The cantlooQUs
prockIctiPn of r1th f~plieUdofeacal_te1alor IlUlpllonill1l
~ts bYpoundilt8~ (~~Iick1ngand 1IkIod1) hOweva ~ t H 1 (
IIIOUld rep1dly altAIr theli11l~t and the rua~ofSRB re
notedtofncreasemiddotttcllll ( 1 SRaIll1taat llaUt 1 x 101 SMIIl-1
aftet bull -th
IUctotJlIIl corrosion canfe=t _ta1sln a1Ullberof Ways
(Iverson1972 fUlllIt end King 1975f 1Uilr 1981) pH can
aff~ the ectivity of the 1IIin organlasinuolved the H 1IIho
hava tin qlu pH range beban 10 12 (Graff 1981)
although tIdS -can be lnf1u8nCed by ather fllltors In atI1tl11n
they lIKIJlre a catbon end nitrogen aurCa 11 various
salts tin alactron accaptor (HedJart ~ idbullbull 1985)bullAil of
U- facton are availabll in the debris -alltting b41tween
ar1na foullng -d the 1lUbstra~ 110 providing bull potantlally
corrosiveanvirorwent
Evan IIIhete such accullIUlatiori does not take place varlable
changas inoxYQ8ll and pH ~trations acrossthe suface of
teel soch as C3I becaustd by foul~ ~ C3I give ti_
to corms1an calla lalildlng to piftlng corrosion
There is an initial tllld8llCY for the pH to rise as a relUlt of
the excretoryactivi1i~ of fouling aniIIal and this 1amp UIIllkely
to give rise to corros1an problems Eventually ~er feacal
h
1
~I
materials and detrltus~ill build up around or~anisms ~re bo~
low pH and p02 devul9ls providing a potentially coJJOampiIu
envinx-nt flaw does dify thh _build UfJ but inmiddotthbull
Ixparlunts carrllldotttid notprlvl(lt it R~~r it
l8Iilated the depth of deposit ~1~~ in t- ~ It ahoutd alao r~~- in f~ population so-1QIJvlWela 111111 be dying or dUd Thlb
~lng~wUl~1Ielacal1Md conoa1ve ~iccandltiorJs-t SR8a 111111 b4I fCllllfC314lng ~y pOIi1desa hlbitt~ HtInd 1tt~ WIi~ul create 1~1Mct Jrcx-ntal dlffarenca ~ ~th l1kel1haOd or pitting
n
References
Albilnder J~T 1 bL~1953~~(9EthII prockIctiPn of IdcroaltJodIIs UIIId in I~ studies Rev Sc1 lnstra ~ 528-531
_r1can Ptbl1c Health laSoc1ation 1975 5taIIdIrd~for the lIIIIIlystS of _tars end taatms 11lth pounddIt1on 1IPHIi-~ 1015 ~taenthSt NW 1IIa-l~~OC 2IJ]J6 USA
Bau-8laquo=k1ng C~IIIG amp 1IIoOd EJF1955 Biologlcal ~ in theestuarlna envi~t 1 EcolClil) of the ~ cycle Kcnlridlkjl Ned Akad -t ~ Ser B~ fi 160-181
cter DB amp SUver IA 1961 In IRafaranctlElectrodl$l EdltolS Iva DlG and l-z Gl lc8dilll1c Pr London Maw Yodlt Chapter 11 ~
Davies PIII I 8r1ric middotF 1942JII1c1oel4iitrodas for-aln local oxygen tension in n11 tissues Rlv Sci InStruII Yo 524-533
EdyileariRJGJ ~ Tetty LA UJ83 The 1nfluenc ~lIlcro algae on corrosion ofstructural steels used 4l~ the 1tIirth
Sea 81odeterioratian sect 336-347
bull bull
~
bull - -- ~It Jf bull
- ~ 13
12
Terry LA I~Ed)VeangtRfiJ 1981 lIi(toal~IOI1CGrmianF~ l)i 1St1oJhemailnEifooiiiigffixed gftshoreIt Bot 1Iar ~ 117-183 lsect~~jlaUoQ1l1 _ ~t Ef)e~gy Offs~r1I Te~lOtjjy Paj)8r
-middotltcntpI)~it$t~l91-16PP~middotmiddotdl ( I
bullbull
iIIalk e1 JL 1971 bull Ion specfie Uquid I~~ e)(charig~ ndcroelactroclels Anal ChIIaI ~8S-Q2 AGraf1~ w~S 9S1~ 1orrosloh InFIntroooctI00 tooffshOle
51~weSoes~~fab~lcat~onIns~allat1on Gulf Wall PI aelUnger EG 1982 The appl1~tlon 9fMl1ikIngmiddotHoust(jfW~Q23g-258middot icroelectrade in the study of pH end PD2 in lIHinli
HerbierC SN fGH~rt~ )RI -Stoctcdalpii Watltif1Sal RS ~1ronIIInb Ub~~ 27l-Z79 1985 Fac~or lontro1l11)9 tha activity ofmiddot sulpMte
bull Wall P 1983 The appllcatlon Md ~ ofiwrJ)e~c~odl shy-taicfii9 tiiCtetili tn~ diiilncj tar infection
techniques for the iiaurnt cif icraeov1ror1liltital5QiCiety ofJebo1- Engi~ SPE 139781-10 _i~~ - ~ ~ i ( _ IKIdif1catlons caued by aGII8 rlnefou~lng -C~ebullbull
PhD lhaaia Ibiverslty or ~tar UKbull bullbull bull -
IIH1nke JA 19fiSa In EXperllRants in Physiology and middotampioClIiIiiilatJift l c E(If(orltGAiKetkutgt ~c-Presa-
Wooltdngton AObullbull Davenport J 1983 pH lind IlIl2 IMla ffl~- j~t~5 -lMIIleath 1IH1na ro-fauUngGqWl J bull Expbull 1
B101 Earlsectsect 113-124 Klnke JAIII 1969b II) Glass IUcroelectJQdes poundx1tors Uwal1~Ui~ OrOllt arid~t Nt JOhn fII11ey YOI1lt thaptar 11
~-f _f~ ~ ~~nt ~ Igt
bull Iverson IIIP 1972 Biolog~ Qrroaian In Advencesin
corrosion scianca and tecIYIology 2EdS FCntaia G~ I bull IIS~et RIIIbull p~ PJaSsLancIoIl 1-42
bull
Jorgensen BB Reusbech NP I Cohen T 19B3 Photosynthesis and strUcture of bIIn~ II1crobWp MC1OIIlectrocki and ~ stud1ea of four cyanabacledalt -~usllWtOl ~bullbull 1~f093
t l~ O~ ~~~ H~ ~9s7~c-gtHatste~iecfn1k van shy pellediftlrtenPt - Stlchelektraden (1-Su
AiJBitiKlificfiliilislflilr)zlIr polrapusdiailJIIissui1g_ liIserstoff(jruQkas fur d1eB lIsUbullbullung der IIliJlttOUf1cillatlon PflftgaHArdI ~fl39
bullbull Mller JOA 1981 tals In EconcinI1c II1croblolooy sect McNbUdbicldatariOtationEd Rosebull A middotAcadelc Praulondcn149-202
t Mller J DAI King RA 1975 8io$~iorat1on of lllltals In lIIi~1al ClflPlCts of the deterloration of tarWbullbull EdsGilbertRJ~ middotloveloCk 0111 bullAcadeic- Presst IILandon 8Jo103
bullt
IPotgate JR 1976 The sulphate-reducingbacteda
Callbridge tIn1versity Press Cliilbrldg8U-K
Reusbecb fUI i Ward DIII 1964 i~~~1Ida5tud1el of
bullt inter$Utilll IIIItez dttm1stry and photosynthetk activity
in ilflaI spring Idcrob1altAppl~EIi~ MerobW tsect4IlU-27lt I
IIbullt bullt
bullbull Ibullt
1
Inmiddot middotmiddot11middotmiddot f~~ a~ -tmiddot bull J~f
I~ W
i5iImiddot J~a t
ilmiddotmiddotmiddot I middotIt Ii
l
~
middotmiddot~~middotlL= g r
Lj Ic
t
i ~
I
gi II cEi ~ I I
1 t rW
D
tR bull
bull t 10
11
r
i
I ~
j
~
I
~bullbullbullbullbullbullbullbullbullbullbull
LV
FigS pH and p02 profiles into macrofoltling eomlllllllitilS dominated by Balm trenatU$ on steel plate 11 two pH profiles Ib) two lIOai profiles leI pH and P02 profiles into dead barllllCle shiella withdetri~Us I I
itIbl r p02 ~ I
nvnHg 11 70
j ifJi eo 1amp0
laquof it 30 s
I 20 N
ca 11 10 a o -- shy i bull
11
80
19
781 o 2 6 8 10 12 0 2 6 amiddot 10 12 dIpdI Qf ~iftto fouling In HighIst numbIII pIatIi
8deg~~
~i c5 iioo5 fiil
pH 8 bull_ ~
~~ ~
_ bullbullbull- pH
-pOz
(cJ
e-Clf ( j~JJlaquo1- --~I ~a ilil - _ I i ]1
~ ri en i ~ oJt ~iJ id tUI
2u1U I shyI t
shy1 2 3 6 6 7 8 9 10 11 12 i shyDepth 9f electrode (mm) into fouling
1 i
1
~----------~-------
~
~-~ lt1~
middotmiddotPmiddot ~
I
V
~
ill
1 Tbellitt~~ of cathodiC protection OO~~lIic~tlO aridrfOflllUlc
ofi AA~Uoullr14l ~1~I
I
ael~lner EG middotIUtcb8l1AJ Dept lt4 lInY1roMe1lal Bl01011
=~~venilt7 IlariCheW 1f139PL $ ~11al1df
r
I
Ablitrict ~~~ e~l billttI ~ted with ~pert1n ant11_1111ampp paint llere itpoii~1n _vatI- In the tl~ld iUIit 1bOioatbrt~ScM Were
1IUbjll~ to qtbocJ~C ~teltt1on byen _ current (~)SJi inci EDAX Iieamp~ta IJdlctecI ~ lachln teoc copper OIl tbePlCtec~ pa1n~ atter14 dQ1a~ protected Pllnt abo ebowed
) reduCed bacteiial activIty butmiddot ~e4 dltoe Iu1d protOzoanmiddot co~~l~tlont~1l IlU l~byenCOCltact not byen dUfwIl00 ~r _baRl lrillolvedln tOXln nil ereio noted
l ~~
1
I
Introduction
Applications of protective coatings and cathodic protect1on(cp~l
are recognised corrosion control methods for IIIBrine steel Itructureso Cathod~c protectlOll preventl conCIsion by Int~lng til8ctrl~l
currentS trem externalllClUfCa to e~~tthlJno~ e~ carrollon feetlans C04U lUChu tUauJ1ng Aelnts a
(
barrIer to ~ the flao of couos1a1 ~aboIHnu~r
Dccurrlng anodes cathodes or wlwit gelvWc CCIIII1 ( 19681
flinger 1980) AnUfauling ~ also JJICOqIOrII~ b1oc~~
Tr~ylt1n nuor~ (~) end o~ Oxide (~) (~ flJ1J
- of u blacld1111~
V _Ut Evans 1981) ~ ~
end growth of bipfoullng 0I1I)IIIII1IIIIe Wdd~ ca ~ of
paint rusendmiddotmanemiddot ~(r~ til )
As corrosian ~t1on ~ bICCIIIII -an ~1tlQd ~ of
antiroullnQs 1 conjunction _ith CP have bllClIOr ~
resultIng in reseudt 1rwestllj111tlng trw COIIIPIitlbUlty or VU~ ~t t _~
rOratlation to dlfrert levI ot CP (~Eelf 18110 ~
~t 1980) RelatIvely 1nIestiljlllUans have ~ urtw ~
or CP an thIt anUraullngpert~ of palnta
In thIt preseot 1nlIestioaUon tV~iln111 ronutlon ud
cOIIIHrclallY incanjlntlon wIth CP was ~t4IC~ Iw I sraed current (-966 l1li) arid exposed at the 1 Strait IInQlbullbullbully
~t1ons weremiddot 00 the IIhcIrt bnifects the PprIUd_
had an a) the various COIIIPQI1eIIts of n pe1ntf1l11 end b) l1oft~
developaentbull
bull bull ~
~ ~ - f
bull ~ _T~~-middot~ ~~- ~r _ -$1
(1) ~rlttionor Painted Steel Billets middot_ ~~tmiddot_tlJ middot~hmiddot~--~fmiddot ~ middot~A~-~~middot - -~ ~
Thirty-al ateel blllets (8-10aD2 end l depth) were cut frollbullt ~middot~middot~middotmiddot~middottrSmiddott ~jS-~ It bull gt~ tfl - middot--imiddotmiddot ~ t1dItrd steal plates The dimenSlonsof -thB butits-lim chosento
bull -~~ middotmiddotgt2h ~ ~-Imiddot ~~rt ~- -middoti~~ ni ~~middot1 l middot-~t~~middot _
(a) fecilltate amiddot Ie detrucUve -thxI of prePraUonpdor to ~~_gt gt~ k7i ~_ t~ 1--1- _ ) - +--_~ -_~bull ming electron ~ 4II1CI (iialloWdirectobs8iVatlCnS of the
- i lt-- ~ gtbull -o1e billet 8Uface
gt r~middotmiddotj ~L(~middot~middot rmiddot tmiddot J ~ - ~ - lt _ bull Single non-standard (1hIula~) nickel lIIireSmiddot Spot Welded e1Cing
bull ~1deJ ~ b~i~ ~j~~ tri afl~1b1ipOi~ f ~1t1 ~4middotmiddot f-gt -~ ~S-- ( -- - - _bull resin EactI billet then Undb~ tI1d prtiladlllltha5O __ eyilr
bull ~~_ ~~imiddot ~1iil h~ intlfcili~ ~eonbt1nlyi Tr1butylt1n F~id(18n)~~~O~dettu20i~blcicld8is 1IIfii bull i - ~ ( then 1IPI)l1ed to glvaadry Ulil tIi1c1ltnOss or 1(1)-150bullbull
tbull (2) IipreSSeCi Current Syat
bull A rigid ~tic due 1I8S drWedllliihwrtv-s41 liiIihOieS IIPProX~
bullbull 05-10 cia episrt ~ Pilnted ampUl8t Wsfbed IntaPQSIUon ampy
threadlng the nickel IIIlr8throu11 the dinto a tarproot junction
box Eliltilen of th8~1l1ii (recelving ~t-d curnritlere IIIlred
Into bull block connectOrrecelvlng dc trOll is potentiOstat The
rainJng iontroi blUets Wet8fbedfl11i1iiy lIIithln thejtlytlonbox
i~ityd~c IkiliPly ~~ ~lat1ruaWiode 8ncIacalOliel
refer~ electtodetoiere fllied ~ the per1aetat of the plut1cdlc IIIId
connected to the pot8ritlostat The desl~ of the potentlosbt ailoiolEld
~PlDteItlO1TClJrnnt to be MIllntained at 8prampset levtl of 966 II1II
c
r~
Grid raf 1J5s-t 114iH 563724 dud 0 Juna
Alolecule andbechr1a in general are ~ to
sutface lI1eroleyer (nlUSton) of _ter (~bullbull 191KJ)~To avold
contaJnatlonof ttw painted surf by U- -vents
blllets Were s-nact in a 1 conte1ner or atc1le (02i ) f~~nd
ter Iih1ch then f9ICI in tv 1IIhere the b~
be r-oved undeNeter El~~ bllleta fa Mcb trtai bullbullel
removed over a 1 dtq (336 hr~ expoeure perioda
each being ~~bn uposurnof 1 5 11 iA 168and DlIIauz1r
Arter field axpauHli the billets wan
fUtered _tar to ~ 1_ debtlflMd far 10 aJnuta ~
wv ~~~endattid sdbUiUd(otO
~ ~l f~ nn1ng el~~ --
lOOted onto 1middotCIi ~bullbullWIsend carbonmiddotcoa~nmiddotmiddot~middot f
each billet Ws~ Iis1ng a ccr1dge ~el~~
(5360) IIIlth anergy dJ8perslve X-ray alcroenelre1a (mx)
_lysis the les Wen reNed end coated III1th vold to al]Ql En
detailed sat of the alaquofa1l
(4) Lfiborattux ~
Painted taa~ bUletalllllre pnparad ~
in a Mrlne reclrculet1ng flOlll _ ~
f1Oll the ~ ~a1t~y Tha~~Un bullybullbullbull
10C M1nta1ned ~~t1as of ~t~la protazOllbull~bull10shy
(Entermap) ~ ~~~middotNlt~Eamp1~ __~and ~ r - ~ -
certaJn ~lc 1IQ1~ 1(1 the
the painted
~
thre repl1catft fn
plKed 1nto(022_)
- thIine1J~~ -lt bull
Atter mAX
(1-2) and i-n-t bn collected
14OC
Pain~
-
12 daY~ exposure and prepared as in d)~ - t
~I ~~~-- i~yen 1~ v
Rflltsi ~ _ ~f-~lI ft j bull _
rt~ll0~~~llY~~~~peln~stetsl~11i~ex~sed for
y~t1~J~ ~~~~1~1~~1UII~ya~kooJ~~~lt ~ ~~_~~I1J~ ~~~tomAX -lysis ~era than gold ~ bull bullbullbull c - _ bull lt
~~fllr ~~~ vllUll ~t1ans of the rfee htures and gt Co
~f~ - ~~~ ~f ~ ~aobta1Mdue IjIll811 iI1Flg 1~2 The IIIIdn ~~ ~ lt)ji9 )tmiddotmiddot - ht
- bull
~I rIIplt~ tIt8 -~ cOlllponanta of the pei1t 1bullbull WelIIlItr1x $ilj~t-~~middot - ~ _ -_
-IUch PVCbQed end rap~13y the chlorlna peak the
i~~~-~~-~V~~~-t18 ~~~~ ~~ Th8
ral~~i~ JlrQP(lrUo1sof ch 1Il-ntal paek~ll1d l~t_ Q~7 -gt- ~ _~ -- gt ) gt -_ -lt ~~ the 0 -end _ xprtsntabullTIwsa changes are -r1sed ~t~ lt~ _ ~ 1nT~ IThe8llia$lb npresentthe chengelI 1n~t1V8 p~lons
ti~~~ [ ~gt 3 - - - _ - _ -
Qf nellllllllt within We surface layars of the pe1ntmiddot ru The
~f~~thtJn -d ~rfeli avr the 336 h11cxI ampScanbe
SII4iIl r~ fig 3 al~ tht rta of~ dirfers tJetwaeo thaa ltmiddot~1- 1ft~- ~ _
IJOl appfiai-ed to ~1nlt1elly until ftar day 168 it fall
~~l~ 1h8relllllrelgn1fi~ diff~be~ qrOtactlid and
proieCted (at -966 ) piIIllnts tor lioo COPPer but not folimiddottin
Ibi~tlons Of the Pain -urface 8hoWad _Pil ~ witilt1M
(Plete I to IV)~ 1hePllat1na (~) pelnt(Phta x and b)
Clearl~st-i~~r alots on the SurfllC8 IiII1kh~ter1sed ~- f hlt1 lriitUl tin conCantrat1on~ lllariy slotsre Partially
~OIIe~ ~ tile _tr~tedal middot1lere rs ~an~ densely packed
grerltr~ ~th 1ar~(iIa1niy cotifIIr) 8ntt -ampniBil (lIB1rily 1rori)being
middot vl~l~~~~~ Dtt~i~~~t~ r~ ~f~~ I~t seawatllr for 11 n IIIOre slots appeared and th~ ~~ess _trb cwer
--if -- V~ c~~ f(i ~~~i~
(Plate Ie to It)~ the granules dld not protrude from the~rtace in ~ ~~~~j1~r~-ffJmiddot0~~
suhap~~r Both 0 nd --960 at ~~pa1nts ~ - fr ~~~~-w1-U~ ~~t
sl~llrat thi stag~ (Plates Ic-d II1d xrnspac~IlYh middotmiddotWith middot middotaJ~cmiddot gt incraallO ~alolre t~ difrrqcbullbull middotbatIiI8IImiddot the protected fjd
bull ~ ~ ~t~
IIllrotacbldpaint ~~lIfIt~ftitr~ h alQt~ p~t bull u bullbullbull J ~~i~~ 1~~
lIIith IIIuper lIdguon u 4lrotatad piIJnt (Pl8ti1 II) 1hD1II8S
Ill )1IMdw1lillS gnnJla~on f-r~~ dpert1cl ~ - - - ~~r ~t
Onthe 966 1IIpetnt t-var tIIO typeaofan with diffannt c t
~hIId~(Plate UI a to c) One hIId tIII-1 bull bullbull e _ bull
fUll wJ1hleas abvlaus alats the o1fwl bed C18UU1IlOt ~litUi middotC c ~ ~
less ~ t11 h Gnnulatlan __ but ~lIiIZly def1nlld in
middot~ latter Afte fila bed tiIIrtIId 10 ~are ~~j(
extansll(tlly ~ the~tIId(O at) tIIIIple lIIIdngthe~ - ~
middot orarallet1ai leas claaI (Plate I~) f u II1ao CQ1tJAJ11d to ~Gn - -
the -966 at 1 (Plata III d tor but 1JI8V8n 1nittNdcneabull -
Althatqa -middotslots wrs abvlaus becaI1ng 1 cwampr aiI- the orwWat1an8
f1nailyftar 14 day exposure ~ fUll drMIlapIIIent on all la - ~
are extensive with the 0 lhawlng both alota pore ~
OVIKgrOloln (Plata II c to f) Gnrulatlon 1bl to ~ gt~t~~ i
or the th1cknats of tho ma ~the ~ at ~ (~~~~ themiddot - - _ middott ~ ~ ~ - -1 1 t
middot f11bull IIIIIS IIOre extans1 thIIIn at dIlya but IIIIIIIY bull1Q~ tI st11l ~w -middotr~ 1gt ~1 ~
obvious fIIany ~ loI8re still p~ yen~~~~~~~r
than before Poiu appearedto be f~lf~ila~~c-~~~a (Plate IVf~ label 1) IIIIl1ch broke~i(2)~ ~~~~
t t
bull ~lS (3) ~ rflC~~ ~i~t~~l~~KI~ t~ldiIiiicyul~loIiih ~~Rht ~ )~- -~ - ~middotd 40 ( gt ~gt ~d lt1 t ~ Jt ~remiddot4amp tha paint trix cavity bottom eroded away reVe81~ng ore
t ~~t~ por~~ ~l~ P~]Y p~~8dby di~lutiaA~ ~~~~ti~Jr~gt~ta(l~~Siinll1cl1Ud~f iUiIiI reriatlGllbullt
bull a~~ ba~tid colCriAtloriccic~ GIl ~~~d ri -966 - ib~~~amp catQn1iti~~t~I~~(~t an ( ~
t the ~~~palnts (PIa_taiiI d~~h ~wh ~ n td have ~ CamplluJMitiIVc) ~~middottti o~lgtnalrf tlt of hIght
~ l=~A ~J~rW tYtles~ p~fiiilriQcOCCold to~Ja to jU-t ~jmiddoti~ai dlatClilS ~ir -jlati ~tbullbullent aNit ~ t ~~ th8middotf~ld~ies~~t~on~ ~ -966 prQ~~~(P14ta Iii~)Omiddotiliof~m1hiiHAiiipieat ere~ll-(piitir~ p-iot~iilans(PiA-iII~ end c) nbullbulltiledt dl~by1i~~ end wirinali1sheii ~cMI~bY thI tili
bull In t-~~ ~1i1nt 1oIIIeHfd~ ~m1IS (15OClind a t ri~r-dlatoa ~lor P~t dlataiOlonlt1anGIl ttw-966
t
bull ~
pr~t~ ~is~ih iion obitJauIJ A ~rot -=1 ~ ~ i __
reprasenttild on the 966 painted 1iUrfc1ncluicunljj ~ rwt
Achnanths Colonisation by bacteria 1n the laboretory bsed
bull eliperlMnt tollMct sbiUar pattern to the fild ~
bull bull Dilicueii
bull bull DUt 1Iidnt ~lona Mve dlttllrint ~~ ictian
It ~ JntM l1tenture ttt Iff yen __ JIll the
bull ~ bicins ani 111 ~t INchlnil (PhillIPs 1t73t EWene~ 1981
In ci1ntlHt tJWY ~tit ~Wl1c iIuCh as 1rbiltyl tirI
~ lite wAlII]ltiClr dlsP8tfu~ middotthiirsttilll~intaJn~ a
bull ~
lIIifoflll ampalCentraUon through the filII 10 thoIt biocide rel_ at tha
paint surfce 1fI$Jlts in IIIOre ditfusing ~s to repla~lj -iflJ~it
resulting 111 bullbulltNdy lr leechlng rata M1llat this IIIIY be middottruIi ot
tin ~ it was nOt true ~ ttw middotlBlf fOZWJlatiGn ua ~ bull - i - bull 1(1
ObSfvt1onundr SE clarly ~ tM t1l1 to be present
~ly lIS OIIIdle-llk parUc1~~t tn lUlfiice (Ito
IV) bUt usa ~ the dIpth attt fU-middot ~y~
If one considra the concn~rtlonmiddot chMge 111 1n ~111c COKICIl8rlts fro- the paint tU- i bullbullbull tMWei b~ ~ Si6t arid
tI-~t iron oxidt 1~ can bt (Fl_ 3 IrId TIibl8I) ~t ~ ase cona1derllble d1fftrenC8s ~ a Un decz bcdYMr ttw p1iod IIIIwraS lron~- an unt Ught inCr h ~ incqJper ~tlGn occUrnd _idly at ttw ~~ Gfb ~ period ~ out ttwn fu nPldly ampQII1n ~ umiddotirid II phast This IIIOUld bt tIlq)8Ctad It cantact l-=h1ng we occun1ng
IIltfIOS8d pertlcl of copper would lMch rapldly on t-n1on 1n - shy JIsewtr There would thlll1 ba bull J-a a1iltUy leu avalUba
i1 ~ ~
_tedal becfM uposed Thi tIOUld ttwn IIllCh Thla pulSing efttct
IIIOUld be expctad to -oth OIItlllith U- putic1 __ -lllible II tot 1~ GIl a on r-x _lao Thla teet 01 put1c1e 1tUng
can tit GIl the lIUrlace ClIlC8 it ~ a pan 1amp t
a 11aM bet in Plat I to IV IIaIrIg _ lit la_ II ~ 0- ~s _- PIate IW $llldl ~bull nshyot poRI flllNltttn Al~ tin ~ lou rete thtn I bullbullbull atill Ivwnce of bull al~t pulsed 1Itue rrlict tII1IIWd
elqlCtaG inIl ttwauUr ~ I I I
0
~ i
~ ~
~I J
bullbullt bullbullbullbullt bull bull ~
bull bull
I
bull t
~~
The same seqiHllle of cevity follliltloii (intttecaseofthetestjjltle
slot- shaped) fClliOileciby SecOlld8rYtlolesfotlllingin thll cavity
bottollis releasing further deep8rtox1ils (Plate XV d and g) As
cavities_re ProdICed ~ tiecaaIelnvadedby_ter~ onelilauld
expect 1ncreaa 1n1IOdha and dIladne lMls t the t1met
which the toxlM uw ~ Ths fCMld tCtHi eo (Flvmiddot 3)
~rttno the vi that_tat ~t1on otthe~ ~lY
il1Cl1l8S8S Idth UIiJ
Differences f~in toxin li~~to Ini 966 ill 8811P~ F ~ bOth Un end ~ lcMIr levels afillW
IIIIIlfI ptaInt(~1~~1ntha~~ ~~f~~ cst rklld with tbiIccpper ElJIc~l ta~Uona invalvinv
w pl8cId at a~ht~~middot3lO~lth Un cqJpel bull
(-960)10 ttW ~af this expei1tIIri1ts thIsiorelfJltdt -
that cqlp8r b8IiIId ~ IIiIOUldProc-ln f41ip1dly t thellllgllitiv
patentW _lied () as 1t 11 further f~1t ~ witt Un
III11ch is u-tat aqulllbd Idth
it If ttda 18 eo~ ~
~ )
frOll1 paintscould bl anqnced by an illprd current clithodic
protecUansysteipos1bly ahart_nino te 11fe of the paInts
antifouling ability Bacterial coloni Uon IIIU rtdUCed on the -s66 paint This
could have been due to unflSllOUlllble electrostatic cqrge on th surtece Th1a effect CIIl pley en IIIportant role in the t~of
bacteria to alrraces (lIIarsMll19E1O) As bacteria appear to IhiJw
greater toxicltyto copper (DMpsey 1981) it is Klre likely that
enhanced cqJper leacll1nl On the -966 AIIIPl c8used the recluCed
activity The IDlol8r toxicity ofUn to bacteria (as reported by
~ -
DelIIPsey Hle1gt or its lowes leaching rat was sUpPorted bythl
relatively rapid recolonisation of the slots by lIIicroorganiSlllS ~ftar thE inlt1al aurge of leachi~ Thailllpressecl current dto ~~ the oppoSite _ff~middot ~r on dtta colonstJon l1lI-1- in the
JIbcJrtory~ta larger rUDIIra f~ fit pro~td ~ther than uriprotected Pafnta
Ccnlus1one
1 tiild-a 18 presaIIUd ~ thattJn~ ~~
~llNChng notd1rf~ ~
2 TIleuseofCP~ ~CUIftnt(at) on1h1a ~fca41ng ~- ~
pa1I]t~ to ~_lAIIach1ngn_ at toxiN - -
3 leIJstaPidbectar1al ca1on1aatJon acaampmId an ~bId Pt1ntJ ~
4 JItore larger alcrofauJing (Oiat_ and PzOtoaoana) Gr~_
occurred on the protcbld~
I l
I
bull bull ~
mfEflpoundaS middot~-t9i _jgt- ~ _ _ ~sev M~J~ (1981) (cilonJzatiooof alUf~lng lnts by _rine
Jbull ~teda9g~1ta IIarina XXlV~ 185-191
- - gt 0 Ie 1 _ - _
poundV8I$i I~lf (1~P~h 1II8rin~Ugae and Foul lng AReviw ith
t p~l-i9Jrlr ~iiMdilto5hiP-fQUllfCiH 8ot~ ~r~ ~~~ XXIVbull lIPwl~J11A
Fotteth CN~R ~kk8nGB IWPIR1seJCiiiroaic1lihd ~e ~JIIth CII Offshore St1ucturu ElllS ~tjI~f~1Dt19i4
- - ~
UKC (1976) I~races ill ~W Ecology KIuverd Univbullbull middotraabullf~iddge ~bull C~~jII illt)Ia(t RC (1980) totinllsa Cattod1CPrOtect1cinbull middotmiddot8AtEmiddot~6haOCIlfif~ T~ p No 108 ~ 18 ~ 1IIHf bull
t bull lt ~J~~~ __ ~
Norkr4irulB(198D) Advanculn UcrOfj~lpound~ltiOyVol 4 (AlItxlaquoIder fl Ed) PlerIw Press lcxIdon~ lIP~ 51-85 ~ l~l - bull bull
PJijllt~i At(fm)~mr~~JnI~1fqul1nQ JlalntlJ Riumh ~JnOrpnic toatJngs 2 192middot
)~J gt0 c ~lt -- ~ __ RogerS~ Ut (1968) ina ~(lnt~fbItliP4ili GI COnoslon ~a Technology)bull bull
Sl~iorr~ VP~Ranald~ Rctd~~ (1S80)_ ~ Experllattal Stud1aiRelate middotEffect or Cathod1c Protict1Oi withmiddot terta1nr1c
Coating Syst~ 12th Annualofr T8Id1na1ogy Co1fItHnC8 liauston TIIlt p 9-12
j
fIG1 sPct FromMon IT8TJCuZOI Int Lliu EOAX
FlG~ Fro TlTCu20Ar-4 Dy Field Ellposult I~ir
il
I c
R hi -00amp=7
-I I I I I I I I I f
iii
bull bull
bullbullbull
I J
J
I Figur 3 Thl chan In reltl rqpoIQnlof -ariOtimiddotjOmponents of the paint film with time Thl vtic~1 ampkiS unl till _n ofmiddotmt Illment per unit OJ fe 50all Cl Na
Cu StI Feeshy
PlistinllFe S7~J6 704 3~74 908
f
r e ~ ee~
middotie ~lt - ~ shybullt bull ~
t -~ l Cu~ o ~ f
t --- 3igt - bull 0 J Sn
cc
t ~
~
bullo
~ bull e
bullbullbullbull bull ~
bull ebull a QI 1- bull 0 40 431282 316
101 dmllIII)bullt bullbullbullt
hr 0 ~~bulla 5~16 ~~ -~~ i166 4S i83 8~1B
24tu 6~480 SD~83
111middot 1~1 OS6
~ 5114 6~69 -51 UIamp middot016- lt ~rmiddot
1d o Mea 818 21 021
~Z21 199 1~1Qmiddot ~Jgt2 058
14 d o ~~ 804 Q~B8~~~a~~t~middot 685
866 372amp 8amp4 OS _i~11[ ~~64
Tttile 1 A81UvlL pzQJJortlcw 0 vxl~oI ~~bull Jn 1I1Dtitctel lt-1166) end lqIlDteCt6 (O)middot~af~_~middot_lodEDf~ middotmiddotmiddotmiddotmiddotmiddotmiddotmiddotmiddotc
~ ~
_----I~
J
I bull
I
J bull tmiddot
7o CONQRESOIMTERNACIONAL
I D~CORRO$IOr(IAAINAEmiddot~USTACIONES~v c)
t 7thflfERHATlONAL CONGRESS
ON IARINE CORROSION AND fO(JUNG
~CONGRE$INtERNATIONAL DE~CORR~ON ~E ET QES~SAUSSURE$
i t
~ ~
bullt
t t
UNIVERSIDAD PourtcNlCAVollncio7J1 NiwiemIif~ 1988
(SPAN
SECcl6lfn
BiOIogro marino Morine biologY ~1i1orine
Theunpub1ishedJllBIlu8criptEl have been reprod~d asr~ielved from the
authors The SCientiUc COlIIIBittee take not respoi~ulbll~~Y for any shy
error or omssi9n
Laa ponenciasno han side publicadas y estanreproducidas tal y como
sellan recibido demiddot BUSmiddot autorea pound1 Conte Cientfico middotno se responsemiddot
bil1za de cualquier error ~ om1si~n
bullbull _ bullbull_ bullbull ___ bull $_ _ L _ - _ --- -__
SESSION II Marine biology
Friday 11th November
SURFACE PROPERTIES ON NONmiddotTOXIC ANTIFOULING PAINT FILM
K Nanishi M Murase Y Yonehara M Kishihara and T Hirama
MICROFOULING ON ALUMINIUM PANELS PLACED IN THE TROPICAL WATERS
N B Bhosle K Venkat and A B Wagh
EFFECT OF TRIBUTYLTIN OXIDE (TBTO) ON THE OVARIAN MATURATION OF THE PRAWN CARIDINA WEBERI
R Nagabhushanam B Indira R Sarojini and Mary Frances Thompson
EFFECT OF BIOACTIVE MATERIALS FROM SOFT CORALS ON BACTERIA ASSOCIATED WITH COMMON FOULER
Vitalina Mary Avelin Mary R Sarojini and R Nagabhushanam
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
-lt 1
l~to the_ batnacl~ 1ls~ COC)trol - fmiddotmiddotmiddot
recod4l9sof W pH 8I)d pOi at the SIJIIEdepth outside tOe 5hell middot-middotvmiddotmiddotmiddot_I~ lgt~middot middotmiddot~_middotmiddoti 1gt )
wete made
-flesolts ~
(a) pH IIIldIDz p~lllllS
The _re~ts for Hancl p02proflle tluough both algal -d - ~ gt Comiddotmiddot i -
eniIIIalfou11ng are gi~ in Figs Sto8bull --
TjpicalProtUeis daIiilttuougha t of Entronlorpha are ~1n -bull l J ltl ~~ -
Fig 5 - lIhllsttyp1cal d1innal fluctuations CICCUIJ8d during bull ~i bull bull e bull bull
photosynthetiC- II1d non-phOtinwnthet1c lods (Fig Sa) mad _~ d q _ _ __ _ i bull
decreases inpHlIIIIl1I~ IIIIa) fro-theaul-f1lCe of ttWt bullbull bull ~ bull
dcyenI tqwards the abstratacn tdch it enclOSlild (fig 511)
~fal1s1n pH--nted to as-ach as 2 units Over 9 IIIj
dIpth
figure Sa ahoid the thangea in pH -d Pl2 between 11ve barnacles
with depthln a stlat1c envirornent A steady decrease in ~
occurred as the plata IlUrfe~es toiereappro8ched pHWtially
fell on one profile before rislngbut lOSe steadily with daPth
on the second praflle SlIdlar changes are IIt10wI in fig 6b
with depth into the shells of dead barnacles 80th oxygen -d
pH levels were re~at1l1E1ly steadY ~ similar to the 8IIIblent - ~- lt
envlrOlWOt Until the -actual dead shell IIias entered When a
IIIUked drop in both fJ02 and pH CICQ11Tedbull
~llell in rigs rit JllellllUrellt$IIere maltle ~
iOlfillidual IlUs$els withtiRle (Fig 7dlb) In thll first
experisent elactrodes were placed indicated (fig)
arCUld indivicUllalsln genU flawJog ~ter The
resUlts show bull ~tle fIwrease ~ IID2 eMIS bull aiJc hour 1Ucj
The pH~t of the~ II4ttIn ~~atmiddot~
81 (te UlbientpH being 80) but 1GII8 ateIIfIU ~ the
1rtIa1ent aJphon to a6bull 111 iIadIIr ~~thI aff~ of
flOiW 00 local ~ciIndltJans~ the ~
~tadbut tile flow at~ turned Off tar 25i1aur
Jadin thelllddleofthlt ~The nailts g1R
in FlQIb -PD2 IIMtls mae ~y inflQldng CCnd1tJanamiddotbut
fell -zkadly fter alhoit lao w flaws caadawJnit a
~ fall lit 90 alii Levels rapidly ratond onca t IIIIUtchadDli agaJil No ILICh teed varlauon ~
in pH around the eldlalent 1pbn 1iIIflImI a taIdyJncn4IM -
obsmt8d aspec1elly cleM to thIIen1Ml CondltLni around
indivOJa1 ~ lIIhllst of interest are lMa repl_tative
of field conditions fro around groups or clIIIPS The
IlICpIIr1ants were ~ ~ted Wittdn denSe clUIIPs of ala
fouling a plate These r8SIJlts are given 10 figs tie (1Il1Ud
IlUlIS8ls) -d 8b (within byssal thrtiIds) In tatic ccinditJans
amprCUld the _15 u-a1(8$ ~ fellrapldly (45l) reelJ pft
rose sl1rjltly and then feU aftar 2 h llhen flowing condltiors
were resuraed after J hthe ~ rose to the original middotlevels shy
The pH rose lArlcedly at the eltictrodenear to the xhalent
bull bull bull bull bull
bull11
bullbullbullbull
it
I
8
s~ (3) but reriialned stlaadyahd ultilllateiy fell silghtiy (2)
inttie genetal fooling inassbull
~
III1thihthe1JyMli~~ Or~selci~5 IiIfieretilltti($trltus teniis ltO -=IiUlteliter~ ~ ~(Flg 8b) ~ lIIithrrolaw~ cCincentreuoos diCIppea framp1~D lBigtif3S
~bUt toaitnpl~lylt4i 150 IIifIo lIIhfjnficw t8st0ted~middot pH
t wi~1rithi thread ClUIps~lMl1BtO 74 (3) IinIt B~ito
785(2) lri-staUc cinfit1at IoihUst ettn88dgtiofthlt ~
it -flnedf81r1y~at aroOOd17S~ lIIlanf1Dlilwu
rellitoteilpft inltleUyJose (by 04 UI1ts at Ii1ta3) but it theo
bull fall only to startrfs1riq tgi1nattM not theIlXper1ilent~middot
bull Dlseusslont f tMresulblmiddot obta1nIid it ~be cl8lirly sean that f(IIJUngI
bull species vethe abUlty to Gtly IIOdlfy thdr loclmiddotmiddot
bull InIl~ts
bullbull L1t1t end dark period tluctuat1ona in algalts IBre obsarved
end ware lri close wpeeIIIen~ with those reported by Terry end
bull ~
Edyv- (1981) with 1ncnasaln both pH end IID2 llIrJngllttt
traquor1ods lIhilst thlshappened ttM surface of the t pre
bull fU_ts ware frelyprojecting into UJe water -deep intottie
mat nearer to the solid _stratlli whl1lre shading fll1lli light andbull accuIIJlatlonmiddot of deconIposJng ~gallIIBtllrial DCCIINed pH valuesbull fell IIIBrkedly Concurrent wlth this dlQp in pH ilaS II fell inbull
bullbull
9
oxygen Such t~ral and $PiItial va11ations in ~andpft
would certainly qive 11se to corrosion cells middotatmiddot the sUbsttatie
surface ~ly resulting in iorroaien pitting
The r] _tabellc ctivlUee or fau)Jng -=raJrwerttmlbullbull
together with their prapeN1ty to-=ci-1tritla~
tlwlsalves wIll also lIied to 1dcIoenv~ lIOdifktIona
Resplretory activities wU1 tend to ~~ tn-the tft and Input caapanants auch a cinia~nY qatk
1mIertabJatesmiddot ara mlk 1bullbullbull middot the bulk of their nlUagllnoua
wastes ~ excrated as ~ 1heexpftunt ncord~ pDz
end pH cningS amN 1nd1v1Q8l~ ~ ~ri
claady 1nd1cat41 the ~ of _tft~t GIl tt ~ of lnIirollliental changes Inmiddot expr_~ta lllhere no Wlter
--nts occurradthmi a np1d dacreUfi1ll pDz
COIJIdalo be a ~decreae in pH (no a) v fladrlg
condlt1C1ns are r-ed tIwn Ieymiddotmiddotor iIay nat be a 1bIIeIqI1int PH
1nctease dBpwIding ~ the --- ofmiddot iheltr ~ by the
organl1n the cluIIp 0wIgn wui a influenced by the
proxJa1ty of 1nhalent or ~a~ the dlnsity IIId the
tIlJcknes$ of the clullp w111alsa t)IMt wiuence in ~
wetr -N_tbull Hencein thick clampllP aUt and detritus
cCUlllulatlon is are COIIIIiIn ~ theh billa IIIdfbls
restricted Br8IJIIltdow1 gf organtc datrital _ter1als CI3IIIIUIIS
oxygen resulting in recLlcing conii1Uons llleasurlllenta haw
shown that sulphids qulckly build up In such deposits and
sulphate recLlcing bacteria y tie-~ SRBconcentrtlonmiddotin
J -I
II II IJI Ii ill
II
-bull I
11
10
the depOSlts woUld terid to Ile lOW at We start of ~r1ments as
condit1onswere nat favourable (POstgate 1979) The cantlooQUs
prockIctiPn of r1th f~plieUdofeacal_te1alor IlUlpllonill1l
~ts bYpoundilt8~ (~~Iick1ngand 1IkIod1) hOweva ~ t H 1 (
IIIOUld rep1dly altAIr theli11l~t and the rua~ofSRB re
notedtofncreasemiddotttcllll ( 1 SRaIll1taat llaUt 1 x 101 SMIIl-1
aftet bull -th
IUctotJlIIl corrosion canfe=t _ta1sln a1Ullberof Ways
(Iverson1972 fUlllIt end King 1975f 1Uilr 1981) pH can
aff~ the ectivity of the 1IIin organlasinuolved the H 1IIho
hava tin qlu pH range beban 10 12 (Graff 1981)
although tIdS -can be lnf1u8nCed by ather fllltors In atI1tl11n
they lIKIJlre a catbon end nitrogen aurCa 11 various
salts tin alactron accaptor (HedJart ~ idbullbull 1985)bullAil of
U- facton are availabll in the debris -alltting b41tween
ar1na foullng -d the 1lUbstra~ 110 providing bull potantlally
corrosiveanvirorwent
Evan IIIhete such accullIUlatiori does not take place varlable
changas inoxYQ8ll and pH ~trations acrossthe suface of
teel soch as C3I becaustd by foul~ ~ C3I give ti_
to corms1an calla lalildlng to piftlng corrosion
There is an initial tllld8llCY for the pH to rise as a relUlt of
the excretoryactivi1i~ of fouling aniIIal and this 1amp UIIllkely
to give rise to corros1an problems Eventually ~er feacal
h
1
~I
materials and detrltus~ill build up around or~anisms ~re bo~
low pH and p02 devul9ls providing a potentially coJJOampiIu
envinx-nt flaw does dify thh _build UfJ but inmiddotthbull
Ixparlunts carrllldotttid notprlvl(lt it R~~r it
l8Iilated the depth of deposit ~1~~ in t- ~ It ahoutd alao r~~- in f~ population so-1QIJvlWela 111111 be dying or dUd Thlb
~lng~wUl~1Ielacal1Md conoa1ve ~iccandltiorJs-t SR8a 111111 b4I fCllllfC314lng ~y pOIi1desa hlbitt~ HtInd 1tt~ WIi~ul create 1~1Mct Jrcx-ntal dlffarenca ~ ~th l1kel1haOd or pitting
n
References
Albilnder J~T 1 bL~1953~~(9EthII prockIctiPn of IdcroaltJodIIs UIIId in I~ studies Rev Sc1 lnstra ~ 528-531
_r1can Ptbl1c Health laSoc1ation 1975 5taIIdIrd~for the lIIIIIlystS of _tars end taatms 11lth pounddIt1on 1IPHIi-~ 1015 ~taenthSt NW 1IIa-l~~OC 2IJ]J6 USA
Bau-8laquo=k1ng C~IIIG amp 1IIoOd EJF1955 Biologlcal ~ in theestuarlna envi~t 1 EcolClil) of the ~ cycle Kcnlridlkjl Ned Akad -t ~ Ser B~ fi 160-181
cter DB amp SUver IA 1961 In IRafaranctlElectrodl$l EdltolS Iva DlG and l-z Gl lc8dilll1c Pr London Maw Yodlt Chapter 11 ~
Davies PIII I 8r1ric middotF 1942JII1c1oel4iitrodas for-aln local oxygen tension in n11 tissues Rlv Sci InStruII Yo 524-533
EdyileariRJGJ ~ Tetty LA UJ83 The 1nfluenc ~lIlcro algae on corrosion ofstructural steels used 4l~ the 1tIirth
Sea 81odeterioratian sect 336-347
bull bull
~
bull - -- ~It Jf bull
- ~ 13
12
Terry LA I~Ed)VeangtRfiJ 1981 lIi(toal~IOI1CGrmianF~ l)i 1St1oJhemailnEifooiiiigffixed gftshoreIt Bot 1Iar ~ 117-183 lsect~~jlaUoQ1l1 _ ~t Ef)e~gy Offs~r1I Te~lOtjjy Paj)8r
-middotltcntpI)~it$t~l91-16PP~middotmiddotdl ( I
bullbull
iIIalk e1 JL 1971 bull Ion specfie Uquid I~~ e)(charig~ ndcroelactroclels Anal ChIIaI ~8S-Q2 AGraf1~ w~S 9S1~ 1orrosloh InFIntroooctI00 tooffshOle
51~weSoes~~fab~lcat~onIns~allat1on Gulf Wall PI aelUnger EG 1982 The appl1~tlon 9fMl1ikIngmiddotHoust(jfW~Q23g-258middot icroelectrade in the study of pH end PD2 in lIHinli
HerbierC SN fGH~rt~ )RI -Stoctcdalpii Watltif1Sal RS ~1ronIIInb Ub~~ 27l-Z79 1985 Fac~or lontro1l11)9 tha activity ofmiddot sulpMte
bull Wall P 1983 The appllcatlon Md ~ ofiwrJ)e~c~odl shy-taicfii9 tiiCtetili tn~ diiilncj tar infection
techniques for the iiaurnt cif icraeov1ror1liltital5QiCiety ofJebo1- Engi~ SPE 139781-10 _i~~ - ~ ~ i ( _ IKIdif1catlons caued by aGII8 rlnefou~lng -C~ebullbull
PhD lhaaia Ibiverslty or ~tar UKbull bullbull bull -
IIH1nke JA 19fiSa In EXperllRants in Physiology and middotampioClIiIiiilatJift l c E(If(orltGAiKetkutgt ~c-Presa-
Wooltdngton AObullbull Davenport J 1983 pH lind IlIl2 IMla ffl~- j~t~5 -lMIIleath 1IH1na ro-fauUngGqWl J bull Expbull 1
B101 Earlsectsect 113-124 Klnke JAIII 1969b II) Glass IUcroelectJQdes poundx1tors Uwal1~Ui~ OrOllt arid~t Nt JOhn fII11ey YOI1lt thaptar 11
~-f _f~ ~ ~~nt ~ Igt
bull Iverson IIIP 1972 Biolog~ Qrroaian In Advencesin
corrosion scianca and tecIYIology 2EdS FCntaia G~ I bull IIS~et RIIIbull p~ PJaSsLancIoIl 1-42
bull
Jorgensen BB Reusbech NP I Cohen T 19B3 Photosynthesis and strUcture of bIIn~ II1crobWp MC1OIIlectrocki and ~ stud1ea of four cyanabacledalt -~usllWtOl ~bullbull 1~f093
t l~ O~ ~~~ H~ ~9s7~c-gtHatste~iecfn1k van shy pellediftlrtenPt - Stlchelektraden (1-Su
AiJBitiKlificfiliilislflilr)zlIr polrapusdiailJIIissui1g_ liIserstoff(jruQkas fur d1eB lIsUbullbullung der IIliJlttOUf1cillatlon PflftgaHArdI ~fl39
bullbull Mller JOA 1981 tals In EconcinI1c II1croblolooy sect McNbUdbicldatariOtationEd Rosebull A middotAcadelc Praulondcn149-202
t Mller J DAI King RA 1975 8io$~iorat1on of lllltals In lIIi~1al ClflPlCts of the deterloration of tarWbullbull EdsGilbertRJ~ middotloveloCk 0111 bullAcadeic- Presst IILandon 8Jo103
bullt
IPotgate JR 1976 The sulphate-reducingbacteda
Callbridge tIn1versity Press Cliilbrldg8U-K
Reusbecb fUI i Ward DIII 1964 i~~~1Ida5tud1el of
bullt inter$Utilll IIIItez dttm1stry and photosynthetk activity
in ilflaI spring Idcrob1altAppl~EIi~ MerobW tsect4IlU-27lt I
IIbullt bullt
bullbull Ibullt
1
Inmiddot middotmiddot11middotmiddot f~~ a~ -tmiddot bull J~f
I~ W
i5iImiddot J~a t
ilmiddotmiddotmiddot I middotIt Ii
l
~
middotmiddot~~middotlL= g r
Lj Ic
t
i ~
I
gi II cEi ~ I I
1 t rW
D
tR bull
bull t 10
11
r
i
I ~
j
~
I
~bullbullbullbullbullbullbullbullbullbullbull
LV
FigS pH and p02 profiles into macrofoltling eomlllllllitilS dominated by Balm trenatU$ on steel plate 11 two pH profiles Ib) two lIOai profiles leI pH and P02 profiles into dead barllllCle shiella withdetri~Us I I
itIbl r p02 ~ I
nvnHg 11 70
j ifJi eo 1amp0
laquof it 30 s
I 20 N
ca 11 10 a o -- shy i bull
11
80
19
781 o 2 6 8 10 12 0 2 6 amiddot 10 12 dIpdI Qf ~iftto fouling In HighIst numbIII pIatIi
8deg~~
~i c5 iioo5 fiil
pH 8 bull_ ~
~~ ~
_ bullbullbull- pH
-pOz
(cJ
e-Clf ( j~JJlaquo1- --~I ~a ilil - _ I i ]1
~ ri en i ~ oJt ~iJ id tUI
2u1U I shyI t
shy1 2 3 6 6 7 8 9 10 11 12 i shyDepth 9f electrode (mm) into fouling
1 i
1
~----------~-------
~
~-~ lt1~
middotmiddotPmiddot ~
I
V
~
ill
1 Tbellitt~~ of cathodiC protection OO~~lIic~tlO aridrfOflllUlc
ofi AA~Uoullr14l ~1~I
I
ael~lner EG middotIUtcb8l1AJ Dept lt4 lInY1roMe1lal Bl01011
=~~venilt7 IlariCheW 1f139PL $ ~11al1df
r
I
Ablitrict ~~~ e~l billttI ~ted with ~pert1n ant11_1111ampp paint llere itpoii~1n _vatI- In the tl~ld iUIit 1bOioatbrt~ScM Were
1IUbjll~ to qtbocJ~C ~teltt1on byen _ current (~)SJi inci EDAX Iieamp~ta IJdlctecI ~ lachln teoc copper OIl tbePlCtec~ pa1n~ atter14 dQ1a~ protected Pllnt abo ebowed
) reduCed bacteiial activIty butmiddot ~e4 dltoe Iu1d protOzoanmiddot co~~l~tlont~1l IlU l~byenCOCltact not byen dUfwIl00 ~r _baRl lrillolvedln tOXln nil ereio noted
l ~~
1
I
Introduction
Applications of protective coatings and cathodic protect1on(cp~l
are recognised corrosion control methods for IIIBrine steel Itructureso Cathod~c protectlOll preventl conCIsion by Int~lng til8ctrl~l
currentS trem externalllClUfCa to e~~tthlJno~ e~ carrollon feetlans C04U lUChu tUauJ1ng Aelnts a
(
barrIer to ~ the flao of couos1a1 ~aboIHnu~r
Dccurrlng anodes cathodes or wlwit gelvWc CCIIII1 ( 19681
flinger 1980) AnUfauling ~ also JJICOqIOrII~ b1oc~~
Tr~ylt1n nuor~ (~) end o~ Oxide (~) (~ flJ1J
- of u blacld1111~
V _Ut Evans 1981) ~ ~
end growth of bipfoullng 0I1I)IIIII1IIIIe Wdd~ ca ~ of
paint rusendmiddotmanemiddot ~(r~ til )
As corrosian ~t1on ~ bICCIIIII -an ~1tlQd ~ of
antiroullnQs 1 conjunction _ith CP have bllClIOr ~
resultIng in reseudt 1rwestllj111tlng trw COIIIPIitlbUlty or VU~ ~t t _~
rOratlation to dlfrert levI ot CP (~Eelf 18110 ~
~t 1980) RelatIvely 1nIestiljlllUans have ~ urtw ~
or CP an thIt anUraullngpert~ of palnta
In thIt preseot 1nlIestioaUon tV~iln111 ronutlon ud
cOIIIHrclallY incanjlntlon wIth CP was ~t4IC~ Iw I sraed current (-966 l1li) arid exposed at the 1 Strait IInQlbullbullbully
~t1ons weremiddot 00 the IIhcIrt bnifects the PprIUd_
had an a) the various COIIIPQI1eIIts of n pe1ntf1l11 end b) l1oft~
developaentbull
bull bull ~
~ ~ - f
bull ~ _T~~-middot~ ~~- ~r _ -$1
(1) ~rlttionor Painted Steel Billets middot_ ~~tmiddot_tlJ middot~hmiddot~--~fmiddot ~ middot~A~-~~middot - -~ ~
Thirty-al ateel blllets (8-10aD2 end l depth) were cut frollbullt ~middot~middot~middotmiddot~middottrSmiddott ~jS-~ It bull gt~ tfl - middot--imiddotmiddot ~ t1dItrd steal plates The dimenSlonsof -thB butits-lim chosento
bull -~~ middotmiddotgt2h ~ ~-Imiddot ~~rt ~- -middoti~~ ni ~~middot1 l middot-~t~~middot _
(a) fecilltate amiddot Ie detrucUve -thxI of prePraUonpdor to ~~_gt gt~ k7i ~_ t~ 1--1- _ ) - +--_~ -_~bull ming electron ~ 4II1CI (iialloWdirectobs8iVatlCnS of the
- i lt-- ~ gtbull -o1e billet 8Uface
gt r~middotmiddotj ~L(~middot~middot rmiddot tmiddot J ~ - ~ - lt _ bull Single non-standard (1hIula~) nickel lIIireSmiddot Spot Welded e1Cing
bull ~1deJ ~ b~i~ ~j~~ tri afl~1b1ipOi~ f ~1t1 ~4middotmiddot f-gt -~ ~S-- ( -- - - _bull resin EactI billet then Undb~ tI1d prtiladlllltha5O __ eyilr
bull ~~_ ~~imiddot ~1iil h~ intlfcili~ ~eonbt1nlyi Tr1butylt1n F~id(18n)~~~O~dettu20i~blcicld8is 1IIfii bull i - ~ ( then 1IPI)l1ed to glvaadry Ulil tIi1c1ltnOss or 1(1)-150bullbull
tbull (2) IipreSSeCi Current Syat
bull A rigid ~tic due 1I8S drWedllliihwrtv-s41 liiIihOieS IIPProX~
bullbull 05-10 cia episrt ~ Pilnted ampUl8t Wsfbed IntaPQSIUon ampy
threadlng the nickel IIIlr8throu11 the dinto a tarproot junction
box Eliltilen of th8~1l1ii (recelving ~t-d curnritlere IIIlred
Into bull block connectOrrecelvlng dc trOll is potentiOstat The
rainJng iontroi blUets Wet8fbedfl11i1iiy lIIithln thejtlytlonbox
i~ityd~c IkiliPly ~~ ~lat1ruaWiode 8ncIacalOliel
refer~ electtodetoiere fllied ~ the per1aetat of the plut1cdlc IIIId
connected to the pot8ritlostat The desl~ of the potentlosbt ailoiolEld
~PlDteItlO1TClJrnnt to be MIllntained at 8prampset levtl of 966 II1II
c
r~
Grid raf 1J5s-t 114iH 563724 dud 0 Juna
Alolecule andbechr1a in general are ~ to
sutface lI1eroleyer (nlUSton) of _ter (~bullbull 191KJ)~To avold
contaJnatlonof ttw painted surf by U- -vents
blllets Were s-nact in a 1 conte1ner or atc1le (02i ) f~~nd
ter Iih1ch then f9ICI in tv 1IIhere the b~
be r-oved undeNeter El~~ bllleta fa Mcb trtai bullbullel
removed over a 1 dtq (336 hr~ expoeure perioda
each being ~~bn uposurnof 1 5 11 iA 168and DlIIauz1r
Arter field axpauHli the billets wan
fUtered _tar to ~ 1_ debtlflMd far 10 aJnuta ~
wv ~~~endattid sdbUiUd(otO
~ ~l f~ nn1ng el~~ --
lOOted onto 1middotCIi ~bullbullWIsend carbonmiddotcoa~nmiddotmiddot~middot f
each billet Ws~ Iis1ng a ccr1dge ~el~~
(5360) IIIlth anergy dJ8perslve X-ray alcroenelre1a (mx)
_lysis the les Wen reNed end coated III1th vold to al]Ql En
detailed sat of the alaquofa1l
(4) Lfiborattux ~
Painted taa~ bUletalllllre pnparad ~
in a Mrlne reclrculet1ng flOlll _ ~
f1Oll the ~ ~a1t~y Tha~~Un bullybullbullbull
10C M1nta1ned ~~t1as of ~t~la protazOllbull~bull10shy
(Entermap) ~ ~~~middotNlt~Eamp1~ __~and ~ r - ~ -
certaJn ~lc 1IQ1~ 1(1 the
the painted
~
thre repl1catft fn
plKed 1nto(022_)
- thIine1J~~ -lt bull
Atter mAX
(1-2) and i-n-t bn collected
14OC
Pain~
-
12 daY~ exposure and prepared as in d)~ - t
~I ~~~-- i~yen 1~ v
Rflltsi ~ _ ~f-~lI ft j bull _
rt~ll0~~~llY~~~~peln~stetsl~11i~ex~sed for
y~t1~J~ ~~~~1~1~~1UII~ya~kooJ~~~lt ~ ~~_~~I1J~ ~~~tomAX -lysis ~era than gold ~ bull bullbullbull c - _ bull lt
~~fllr ~~~ vllUll ~t1ans of the rfee htures and gt Co
~f~ - ~~~ ~f ~ ~aobta1Mdue IjIll811 iI1Flg 1~2 The IIIIdn ~~ ~ lt)ji9 )tmiddotmiddot - ht
- bull
~I rIIplt~ tIt8 -~ cOlllponanta of the pei1t 1bullbull WelIIlItr1x $ilj~t-~~middot - ~ _ -_
-IUch PVCbQed end rap~13y the chlorlna peak the
i~~~-~~-~V~~~-t18 ~~~~ ~~ Th8
ral~~i~ JlrQP(lrUo1sof ch 1Il-ntal paek~ll1d l~t_ Q~7 -gt- ~ _~ -- gt ) gt -_ -lt ~~ the 0 -end _ xprtsntabullTIwsa changes are -r1sed ~t~ lt~ _ ~ 1nT~ IThe8llia$lb npresentthe chengelI 1n~t1V8 p~lons
ti~~~ [ ~gt 3 - - - _ - _ -
Qf nellllllllt within We surface layars of the pe1ntmiddot ru The
~f~~thtJn -d ~rfeli avr the 336 h11cxI ampScanbe
SII4iIl r~ fig 3 al~ tht rta of~ dirfers tJetwaeo thaa ltmiddot~1- 1ft~- ~ _
IJOl appfiai-ed to ~1nlt1elly until ftar day 168 it fall
~~l~ 1h8relllllrelgn1fi~ diff~be~ qrOtactlid and
proieCted (at -966 ) piIIllnts tor lioo COPPer but not folimiddottin
Ibi~tlons Of the Pain -urface 8hoWad _Pil ~ witilt1M
(Plete I to IV)~ 1hePllat1na (~) pelnt(Phta x and b)
Clearl~st-i~~r alots on the SurfllC8 IiII1kh~ter1sed ~- f hlt1 lriitUl tin conCantrat1on~ lllariy slotsre Partially
~OIIe~ ~ tile _tr~tedal middot1lere rs ~an~ densely packed
grerltr~ ~th 1ar~(iIa1niy cotifIIr) 8ntt -ampniBil (lIB1rily 1rori)being
middot vl~l~~~~~ Dtt~i~~~t~ r~ ~f~~ I~t seawatllr for 11 n IIIOre slots appeared and th~ ~~ess _trb cwer
--if -- V~ c~~ f(i ~~~i~
(Plate Ie to It)~ the granules dld not protrude from the~rtace in ~ ~~~~j1~r~-ffJmiddot0~~
suhap~~r Both 0 nd --960 at ~~pa1nts ~ - fr ~~~~-w1-U~ ~~t
sl~llrat thi stag~ (Plates Ic-d II1d xrnspac~IlYh middotmiddotWith middot middotaJ~cmiddot gt incraallO ~alolre t~ difrrqcbullbull middotbatIiI8IImiddot the protected fjd
bull ~ ~ ~t~
IIllrotacbldpaint ~~lIfIt~ftitr~ h alQt~ p~t bull u bullbullbull J ~~i~~ 1~~
lIIith IIIuper lIdguon u 4lrotatad piIJnt (Pl8ti1 II) 1hD1II8S
Ill )1IMdw1lillS gnnJla~on f-r~~ dpert1cl ~ - - - ~~r ~t
Onthe 966 1IIpetnt t-var tIIO typeaofan with diffannt c t
~hIId~(Plate UI a to c) One hIId tIII-1 bull bullbull e _ bull
fUll wJ1hleas abvlaus alats the o1fwl bed C18UU1IlOt ~litUi middotC c ~ ~
less ~ t11 h Gnnulatlan __ but ~lIiIZly def1nlld in
middot~ latter Afte fila bed tiIIrtIId 10 ~are ~~j(
extansll(tlly ~ the~tIId(O at) tIIIIple lIIIdngthe~ - ~
middot orarallet1ai leas claaI (Plate I~) f u II1ao CQ1tJAJ11d to ~Gn - -
the -966 at 1 (Plata III d tor but 1JI8V8n 1nittNdcneabull -
Althatqa -middotslots wrs abvlaus becaI1ng 1 cwampr aiI- the orwWat1an8
f1nailyftar 14 day exposure ~ fUll drMIlapIIIent on all la - ~
are extensive with the 0 lhawlng both alota pore ~
OVIKgrOloln (Plata II c to f) Gnrulatlon 1bl to ~ gt~t~~ i
or the th1cknats of tho ma ~the ~ at ~ (~~~~ themiddot - - _ middott ~ ~ ~ - -1 1 t
middot f11bull IIIIIS IIOre extans1 thIIIn at dIlya but IIIIIIIY bull1Q~ tI st11l ~w -middotr~ 1gt ~1 ~
obvious fIIany ~ loI8re still p~ yen~~~~~~~r
than before Poiu appearedto be f~lf~ila~~c-~~~a (Plate IVf~ label 1) IIIIl1ch broke~i(2)~ ~~~~
t t
bull ~lS (3) ~ rflC~~ ~i~t~~l~~KI~ t~ldiIiiicyul~loIiih ~~Rht ~ )~- -~ - ~middotd 40 ( gt ~gt ~d lt1 t ~ Jt ~remiddot4amp tha paint trix cavity bottom eroded away reVe81~ng ore
t ~~t~ por~~ ~l~ P~]Y p~~8dby di~lutiaA~ ~~~~ti~Jr~gt~ta(l~~Siinll1cl1Ud~f iUiIiI reriatlGllbullt
bull a~~ ba~tid colCriAtloriccic~ GIl ~~~d ri -966 - ib~~~amp catQn1iti~~t~I~~(~t an ( ~
t the ~~~palnts (PIa_taiiI d~~h ~wh ~ n td have ~ CamplluJMitiIVc) ~~middottti o~lgtnalrf tlt of hIght
~ l=~A ~J~rW tYtles~ p~fiiilriQcOCCold to~Ja to jU-t ~jmiddoti~ai dlatClilS ~ir -jlati ~tbullbullent aNit ~ t ~~ th8middotf~ld~ies~~t~on~ ~ -966 prQ~~~(P14ta Iii~)Omiddotiliof~m1hiiHAiiipieat ere~ll-(piitir~ p-iot~iilans(PiA-iII~ end c) nbullbulltiledt dl~by1i~~ end wirinali1sheii ~cMI~bY thI tili
bull In t-~~ ~1i1nt 1oIIIeHfd~ ~m1IS (15OClind a t ri~r-dlatoa ~lor P~t dlataiOlonlt1anGIl ttw-966
t
bull ~
pr~t~ ~is~ih iion obitJauIJ A ~rot -=1 ~ ~ i __
reprasenttild on the 966 painted 1iUrfc1ncluicunljj ~ rwt
Achnanths Colonisation by bacteria 1n the laboretory bsed
bull eliperlMnt tollMct sbiUar pattern to the fild ~
bull bull Dilicueii
bull bull DUt 1Iidnt ~lona Mve dlttllrint ~~ ictian
It ~ JntM l1tenture ttt Iff yen __ JIll the
bull ~ bicins ani 111 ~t INchlnil (PhillIPs 1t73t EWene~ 1981
In ci1ntlHt tJWY ~tit ~Wl1c iIuCh as 1rbiltyl tirI
~ lite wAlII]ltiClr dlsP8tfu~ middotthiirsttilll~intaJn~ a
bull ~
lIIifoflll ampalCentraUon through the filII 10 thoIt biocide rel_ at tha
paint surfce 1fI$Jlts in IIIOre ditfusing ~s to repla~lj -iflJ~it
resulting 111 bullbulltNdy lr leechlng rata M1llat this IIIIY be middottruIi ot
tin ~ it was nOt true ~ ttw middotlBlf fOZWJlatiGn ua ~ bull - i - bull 1(1
ObSfvt1onundr SE clarly ~ tM t1l1 to be present
~ly lIS OIIIdle-llk parUc1~~t tn lUlfiice (Ito
IV) bUt usa ~ the dIpth attt fU-middot ~y~
If one considra the concn~rtlonmiddot chMge 111 1n ~111c COKICIl8rlts fro- the paint tU- i bullbullbull tMWei b~ ~ Si6t arid
tI-~t iron oxidt 1~ can bt (Fl_ 3 IrId TIibl8I) ~t ~ ase cona1derllble d1fftrenC8s ~ a Un decz bcdYMr ttw p1iod IIIIwraS lron~- an unt Ught inCr h ~ incqJper ~tlGn occUrnd _idly at ttw ~~ Gfb ~ period ~ out ttwn fu nPldly ampQII1n ~ umiddotirid II phast This IIIOUld bt tIlq)8Ctad It cantact l-=h1ng we occun1ng
IIltfIOS8d pertlcl of copper would lMch rapldly on t-n1on 1n - shy JIsewtr There would thlll1 ba bull J-a a1iltUy leu avalUba
i1 ~ ~
_tedal becfM uposed Thi tIOUld ttwn IIllCh Thla pulSing efttct
IIIOUld be expctad to -oth OIItlllith U- putic1 __ -lllible II tot 1~ GIl a on r-x _lao Thla teet 01 put1c1e 1tUng
can tit GIl the lIUrlace ClIlC8 it ~ a pan 1amp t
a 11aM bet in Plat I to IV IIaIrIg _ lit la_ II ~ 0- ~s _- PIate IW $llldl ~bull nshyot poRI flllNltttn Al~ tin ~ lou rete thtn I bullbullbull atill Ivwnce of bull al~t pulsed 1Itue rrlict tII1IIWd
elqlCtaG inIl ttwauUr ~ I I I
0
~ i
~ ~
~I J
bullbullt bullbullbullbullt bull bull ~
bull bull
I
bull t
~~
The same seqiHllle of cevity follliltloii (intttecaseofthetestjjltle
slot- shaped) fClliOileciby SecOlld8rYtlolesfotlllingin thll cavity
bottollis releasing further deep8rtox1ils (Plate XV d and g) As
cavities_re ProdICed ~ tiecaaIelnvadedby_ter~ onelilauld
expect 1ncreaa 1n1IOdha and dIladne lMls t the t1met
which the toxlM uw ~ Ths fCMld tCtHi eo (Flvmiddot 3)
~rttno the vi that_tat ~t1on otthe~ ~lY
il1Cl1l8S8S Idth UIiJ
Differences f~in toxin li~~to Ini 966 ill 8811P~ F ~ bOth Un end ~ lcMIr levels afillW
IIIIIlfI ptaInt(~1~~1ntha~~ ~~f~~ cst rklld with tbiIccpper ElJIc~l ta~Uona invalvinv
w pl8cId at a~ht~~middot3lO~lth Un cqJpel bull
(-960)10 ttW ~af this expei1tIIri1ts thIsiorelfJltdt -
that cqlp8r b8IiIId ~ IIiIOUldProc-ln f41ip1dly t thellllgllitiv
patentW _lied () as 1t 11 further f~1t ~ witt Un
III11ch is u-tat aqulllbd Idth
it If ttda 18 eo~ ~
~ )
frOll1 paintscould bl anqnced by an illprd current clithodic
protecUansysteipos1bly ahart_nino te 11fe of the paInts
antifouling ability Bacterial coloni Uon IIIU rtdUCed on the -s66 paint This
could have been due to unflSllOUlllble electrostatic cqrge on th surtece Th1a effect CIIl pley en IIIportant role in the t~of
bacteria to alrraces (lIIarsMll19E1O) As bacteria appear to IhiJw
greater toxicltyto copper (DMpsey 1981) it is Klre likely that
enhanced cqJper leacll1nl On the -966 AIIIPl c8used the recluCed
activity The IDlol8r toxicity ofUn to bacteria (as reported by
~ -
DelIIPsey Hle1gt or its lowes leaching rat was sUpPorted bythl
relatively rapid recolonisation of the slots by lIIicroorganiSlllS ~ftar thE inlt1al aurge of leachi~ Thailllpressecl current dto ~~ the oppoSite _ff~middot ~r on dtta colonstJon l1lI-1- in the
JIbcJrtory~ta larger rUDIIra f~ fit pro~td ~ther than uriprotected Pafnta
Ccnlus1one
1 tiild-a 18 presaIIUd ~ thattJn~ ~~
~llNChng notd1rf~ ~
2 TIleuseofCP~ ~CUIftnt(at) on1h1a ~fca41ng ~- ~
pa1I]t~ to ~_lAIIach1ngn_ at toxiN - -
3 leIJstaPidbectar1al ca1on1aatJon acaampmId an ~bId Pt1ntJ ~
4 JItore larger alcrofauJing (Oiat_ and PzOtoaoana) Gr~_
occurred on the protcbld~
I l
I
bull bull ~
mfEflpoundaS middot~-t9i _jgt- ~ _ _ ~sev M~J~ (1981) (cilonJzatiooof alUf~lng lnts by _rine
Jbull ~teda9g~1ta IIarina XXlV~ 185-191
- - gt 0 Ie 1 _ - _
poundV8I$i I~lf (1~P~h 1II8rin~Ugae and Foul lng AReviw ith
t p~l-i9Jrlr ~iiMdilto5hiP-fQUllfCiH 8ot~ ~r~ ~~~ XXIVbull lIPwl~J11A
Fotteth CN~R ~kk8nGB IWPIR1seJCiiiroaic1lihd ~e ~JIIth CII Offshore St1ucturu ElllS ~tjI~f~1Dt19i4
- - ~
UKC (1976) I~races ill ~W Ecology KIuverd Univbullbull middotraabullf~iddge ~bull C~~jII illt)Ia(t RC (1980) totinllsa Cattod1CPrOtect1cinbull middotmiddot8AtEmiddot~6haOCIlfif~ T~ p No 108 ~ 18 ~ 1IIHf bull
t bull lt ~J~~~ __ ~
Norkr4irulB(198D) Advanculn UcrOfj~lpound~ltiOyVol 4 (AlItxlaquoIder fl Ed) PlerIw Press lcxIdon~ lIP~ 51-85 ~ l~l - bull bull
PJijllt~i At(fm)~mr~~JnI~1fqul1nQ JlalntlJ Riumh ~JnOrpnic toatJngs 2 192middot
)~J gt0 c ~lt -- ~ __ RogerS~ Ut (1968) ina ~(lnt~fbItliP4ili GI COnoslon ~a Technology)bull bull
Sl~iorr~ VP~Ranald~ Rctd~~ (1S80)_ ~ Experllattal Stud1aiRelate middotEffect or Cathod1c Protict1Oi withmiddot terta1nr1c
Coating Syst~ 12th Annualofr T8Id1na1ogy Co1fItHnC8 liauston TIIlt p 9-12
j
fIG1 sPct FromMon IT8TJCuZOI Int Lliu EOAX
FlG~ Fro TlTCu20Ar-4 Dy Field Ellposult I~ir
il
I c
R hi -00amp=7
-I I I I I I I I I f
iii
bull bull
bullbullbull
I J
J
I Figur 3 Thl chan In reltl rqpoIQnlof -ariOtimiddotjOmponents of the paint film with time Thl vtic~1 ampkiS unl till _n ofmiddotmt Illment per unit OJ fe 50all Cl Na
Cu StI Feeshy
PlistinllFe S7~J6 704 3~74 908
f
r e ~ ee~
middotie ~lt - ~ shybullt bull ~
t -~ l Cu~ o ~ f
t --- 3igt - bull 0 J Sn
cc
t ~
~
bullo
~ bull e
bullbullbullbull bull ~
bull ebull a QI 1- bull 0 40 431282 316
101 dmllIII)bullt bullbullbullt
hr 0 ~~bulla 5~16 ~~ -~~ i166 4S i83 8~1B
24tu 6~480 SD~83
111middot 1~1 OS6
~ 5114 6~69 -51 UIamp middot016- lt ~rmiddot
1d o Mea 818 21 021
~Z21 199 1~1Qmiddot ~Jgt2 058
14 d o ~~ 804 Q~B8~~~a~~t~middot 685
866 372amp 8amp4 OS _i~11[ ~~64
Tttile 1 A81UvlL pzQJJortlcw 0 vxl~oI ~~bull Jn 1I1Dtitctel lt-1166) end lqIlDteCt6 (O)middot~af~_~middot_lodEDf~ middotmiddotmiddotmiddotmiddotmiddotmiddotmiddotmiddotc
~ ~
_----I~
J
I bull
I
J bull tmiddot
7o CONQRESOIMTERNACIONAL
I D~CORRO$IOr(IAAINAEmiddot~USTACIONES~v c)
t 7thflfERHATlONAL CONGRESS
ON IARINE CORROSION AND fO(JUNG
~CONGRE$INtERNATIONAL DE~CORR~ON ~E ET QES~SAUSSURE$
i t
~ ~
bullt
t t
UNIVERSIDAD PourtcNlCAVollncio7J1 NiwiemIif~ 1988
(SPAN
SECcl6lfn
BiOIogro marino Morine biologY ~1i1orine
Theunpub1ishedJllBIlu8criptEl have been reprod~d asr~ielved from the
authors The SCientiUc COlIIIBittee take not respoi~ulbll~~Y for any shy
error or omssi9n
Laa ponenciasno han side publicadas y estanreproducidas tal y como
sellan recibido demiddot BUSmiddot autorea pound1 Conte Cientfico middotno se responsemiddot
bil1za de cualquier error ~ om1si~n
bullbull _ bullbull_ bullbull ___ bull $_ _ L _ - _ --- -__
SESSION II Marine biology
Friday 11th November
SURFACE PROPERTIES ON NONmiddotTOXIC ANTIFOULING PAINT FILM
K Nanishi M Murase Y Yonehara M Kishihara and T Hirama
MICROFOULING ON ALUMINIUM PANELS PLACED IN THE TROPICAL WATERS
N B Bhosle K Venkat and A B Wagh
EFFECT OF TRIBUTYLTIN OXIDE (TBTO) ON THE OVARIAN MATURATION OF THE PRAWN CARIDINA WEBERI
R Nagabhushanam B Indira R Sarojini and Mary Frances Thompson
EFFECT OF BIOACTIVE MATERIALS FROM SOFT CORALS ON BACTERIA ASSOCIATED WITH COMMON FOULER
Vitalina Mary Avelin Mary R Sarojini and R Nagabhushanam
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
bull bull bull bull bull
bull11
bullbullbullbull
it
I
8
s~ (3) but reriialned stlaadyahd ultilllateiy fell silghtiy (2)
inttie genetal fooling inassbull
~
III1thihthe1JyMli~~ Or~selci~5 IiIfieretilltti($trltus teniis ltO -=IiUlteliter~ ~ ~(Flg 8b) ~ lIIithrrolaw~ cCincentreuoos diCIppea framp1~D lBigtif3S
~bUt toaitnpl~lylt4i 150 IIifIo lIIhfjnficw t8st0ted~middot pH
t wi~1rithi thread ClUIps~lMl1BtO 74 (3) IinIt B~ito
785(2) lri-staUc cinfit1at IoihUst ettn88dgtiofthlt ~
it -flnedf81r1y~at aroOOd17S~ lIIlanf1Dlilwu
rellitoteilpft inltleUyJose (by 04 UI1ts at Ii1ta3) but it theo
bull fall only to startrfs1riq tgi1nattM not theIlXper1ilent~middot
bull Dlseusslont f tMresulblmiddot obta1nIid it ~be cl8lirly sean that f(IIJUngI
bull species vethe abUlty to Gtly IIOdlfy thdr loclmiddotmiddot
bull InIl~ts
bullbull L1t1t end dark period tluctuat1ona in algalts IBre obsarved
end ware lri close wpeeIIIen~ with those reported by Terry end
bull ~
Edyv- (1981) with 1ncnasaln both pH end IID2 llIrJngllttt
traquor1ods lIhilst thlshappened ttM surface of the t pre
bull fU_ts ware frelyprojecting into UJe water -deep intottie
mat nearer to the solid _stratlli whl1lre shading fll1lli light andbull accuIIJlatlonmiddot of deconIposJng ~gallIIBtllrial DCCIINed pH valuesbull fell IIIBrkedly Concurrent wlth this dlQp in pH ilaS II fell inbull
bullbull
9
oxygen Such t~ral and $PiItial va11ations in ~andpft
would certainly qive 11se to corrosion cells middotatmiddot the sUbsttatie
surface ~ly resulting in iorroaien pitting
The r] _tabellc ctivlUee or fau)Jng -=raJrwerttmlbullbull
together with their prapeN1ty to-=ci-1tritla~
tlwlsalves wIll also lIied to 1dcIoenv~ lIOdifktIona
Resplretory activities wU1 tend to ~~ tn-the tft and Input caapanants auch a cinia~nY qatk
1mIertabJatesmiddot ara mlk 1bullbullbull middot the bulk of their nlUagllnoua
wastes ~ excrated as ~ 1heexpftunt ncord~ pDz
end pH cningS amN 1nd1v1Q8l~ ~ ~ri
claady 1nd1cat41 the ~ of _tft~t GIl tt ~ of lnIirollliental changes Inmiddot expr_~ta lllhere no Wlter
--nts occurradthmi a np1d dacreUfi1ll pDz
COIJIdalo be a ~decreae in pH (no a) v fladrlg
condlt1C1ns are r-ed tIwn Ieymiddotmiddotor iIay nat be a 1bIIeIqI1int PH
1nctease dBpwIding ~ the --- ofmiddot iheltr ~ by the
organl1n the cluIIp 0wIgn wui a influenced by the
proxJa1ty of 1nhalent or ~a~ the dlnsity IIId the
tIlJcknes$ of the clullp w111alsa t)IMt wiuence in ~
wetr -N_tbull Hencein thick clampllP aUt and detritus
cCUlllulatlon is are COIIIIiIn ~ theh billa IIIdfbls
restricted Br8IJIIltdow1 gf organtc datrital _ter1als CI3IIIIUIIS
oxygen resulting in recLlcing conii1Uons llleasurlllenta haw
shown that sulphids qulckly build up In such deposits and
sulphate recLlcing bacteria y tie-~ SRBconcentrtlonmiddotin
J -I
II II IJI Ii ill
II
-bull I
11
10
the depOSlts woUld terid to Ile lOW at We start of ~r1ments as
condit1onswere nat favourable (POstgate 1979) The cantlooQUs
prockIctiPn of r1th f~plieUdofeacal_te1alor IlUlpllonill1l
~ts bYpoundilt8~ (~~Iick1ngand 1IkIod1) hOweva ~ t H 1 (
IIIOUld rep1dly altAIr theli11l~t and the rua~ofSRB re
notedtofncreasemiddotttcllll ( 1 SRaIll1taat llaUt 1 x 101 SMIIl-1
aftet bull -th
IUctotJlIIl corrosion canfe=t _ta1sln a1Ullberof Ways
(Iverson1972 fUlllIt end King 1975f 1Uilr 1981) pH can
aff~ the ectivity of the 1IIin organlasinuolved the H 1IIho
hava tin qlu pH range beban 10 12 (Graff 1981)
although tIdS -can be lnf1u8nCed by ather fllltors In atI1tl11n
they lIKIJlre a catbon end nitrogen aurCa 11 various
salts tin alactron accaptor (HedJart ~ idbullbull 1985)bullAil of
U- facton are availabll in the debris -alltting b41tween
ar1na foullng -d the 1lUbstra~ 110 providing bull potantlally
corrosiveanvirorwent
Evan IIIhete such accullIUlatiori does not take place varlable
changas inoxYQ8ll and pH ~trations acrossthe suface of
teel soch as C3I becaustd by foul~ ~ C3I give ti_
to corms1an calla lalildlng to piftlng corrosion
There is an initial tllld8llCY for the pH to rise as a relUlt of
the excretoryactivi1i~ of fouling aniIIal and this 1amp UIIllkely
to give rise to corros1an problems Eventually ~er feacal
h
1
~I
materials and detrltus~ill build up around or~anisms ~re bo~
low pH and p02 devul9ls providing a potentially coJJOampiIu
envinx-nt flaw does dify thh _build UfJ but inmiddotthbull
Ixparlunts carrllldotttid notprlvl(lt it R~~r it
l8Iilated the depth of deposit ~1~~ in t- ~ It ahoutd alao r~~- in f~ population so-1QIJvlWela 111111 be dying or dUd Thlb
~lng~wUl~1Ielacal1Md conoa1ve ~iccandltiorJs-t SR8a 111111 b4I fCllllfC314lng ~y pOIi1desa hlbitt~ HtInd 1tt~ WIi~ul create 1~1Mct Jrcx-ntal dlffarenca ~ ~th l1kel1haOd or pitting
n
References
Albilnder J~T 1 bL~1953~~(9EthII prockIctiPn of IdcroaltJodIIs UIIId in I~ studies Rev Sc1 lnstra ~ 528-531
_r1can Ptbl1c Health laSoc1ation 1975 5taIIdIrd~for the lIIIIIlystS of _tars end taatms 11lth pounddIt1on 1IPHIi-~ 1015 ~taenthSt NW 1IIa-l~~OC 2IJ]J6 USA
Bau-8laquo=k1ng C~IIIG amp 1IIoOd EJF1955 Biologlcal ~ in theestuarlna envi~t 1 EcolClil) of the ~ cycle Kcnlridlkjl Ned Akad -t ~ Ser B~ fi 160-181
cter DB amp SUver IA 1961 In IRafaranctlElectrodl$l EdltolS Iva DlG and l-z Gl lc8dilll1c Pr London Maw Yodlt Chapter 11 ~
Davies PIII I 8r1ric middotF 1942JII1c1oel4iitrodas for-aln local oxygen tension in n11 tissues Rlv Sci InStruII Yo 524-533
EdyileariRJGJ ~ Tetty LA UJ83 The 1nfluenc ~lIlcro algae on corrosion ofstructural steels used 4l~ the 1tIirth
Sea 81odeterioratian sect 336-347
bull bull
~
bull - -- ~It Jf bull
- ~ 13
12
Terry LA I~Ed)VeangtRfiJ 1981 lIi(toal~IOI1CGrmianF~ l)i 1St1oJhemailnEifooiiiigffixed gftshoreIt Bot 1Iar ~ 117-183 lsect~~jlaUoQ1l1 _ ~t Ef)e~gy Offs~r1I Te~lOtjjy Paj)8r
-middotltcntpI)~it$t~l91-16PP~middotmiddotdl ( I
bullbull
iIIalk e1 JL 1971 bull Ion specfie Uquid I~~ e)(charig~ ndcroelactroclels Anal ChIIaI ~8S-Q2 AGraf1~ w~S 9S1~ 1orrosloh InFIntroooctI00 tooffshOle
51~weSoes~~fab~lcat~onIns~allat1on Gulf Wall PI aelUnger EG 1982 The appl1~tlon 9fMl1ikIngmiddotHoust(jfW~Q23g-258middot icroelectrade in the study of pH end PD2 in lIHinli
HerbierC SN fGH~rt~ )RI -Stoctcdalpii Watltif1Sal RS ~1ronIIInb Ub~~ 27l-Z79 1985 Fac~or lontro1l11)9 tha activity ofmiddot sulpMte
bull Wall P 1983 The appllcatlon Md ~ ofiwrJ)e~c~odl shy-taicfii9 tiiCtetili tn~ diiilncj tar infection
techniques for the iiaurnt cif icraeov1ror1liltital5QiCiety ofJebo1- Engi~ SPE 139781-10 _i~~ - ~ ~ i ( _ IKIdif1catlons caued by aGII8 rlnefou~lng -C~ebullbull
PhD lhaaia Ibiverslty or ~tar UKbull bullbull bull -
IIH1nke JA 19fiSa In EXperllRants in Physiology and middotampioClIiIiiilatJift l c E(If(orltGAiKetkutgt ~c-Presa-
Wooltdngton AObullbull Davenport J 1983 pH lind IlIl2 IMla ffl~- j~t~5 -lMIIleath 1IH1na ro-fauUngGqWl J bull Expbull 1
B101 Earlsectsect 113-124 Klnke JAIII 1969b II) Glass IUcroelectJQdes poundx1tors Uwal1~Ui~ OrOllt arid~t Nt JOhn fII11ey YOI1lt thaptar 11
~-f _f~ ~ ~~nt ~ Igt
bull Iverson IIIP 1972 Biolog~ Qrroaian In Advencesin
corrosion scianca and tecIYIology 2EdS FCntaia G~ I bull IIS~et RIIIbull p~ PJaSsLancIoIl 1-42
bull
Jorgensen BB Reusbech NP I Cohen T 19B3 Photosynthesis and strUcture of bIIn~ II1crobWp MC1OIIlectrocki and ~ stud1ea of four cyanabacledalt -~usllWtOl ~bullbull 1~f093
t l~ O~ ~~~ H~ ~9s7~c-gtHatste~iecfn1k van shy pellediftlrtenPt - Stlchelektraden (1-Su
AiJBitiKlificfiliilislflilr)zlIr polrapusdiailJIIissui1g_ liIserstoff(jruQkas fur d1eB lIsUbullbullung der IIliJlttOUf1cillatlon PflftgaHArdI ~fl39
bullbull Mller JOA 1981 tals In EconcinI1c II1croblolooy sect McNbUdbicldatariOtationEd Rosebull A middotAcadelc Praulondcn149-202
t Mller J DAI King RA 1975 8io$~iorat1on of lllltals In lIIi~1al ClflPlCts of the deterloration of tarWbullbull EdsGilbertRJ~ middotloveloCk 0111 bullAcadeic- Presst IILandon 8Jo103
bullt
IPotgate JR 1976 The sulphate-reducingbacteda
Callbridge tIn1versity Press Cliilbrldg8U-K
Reusbecb fUI i Ward DIII 1964 i~~~1Ida5tud1el of
bullt inter$Utilll IIIItez dttm1stry and photosynthetk activity
in ilflaI spring Idcrob1altAppl~EIi~ MerobW tsect4IlU-27lt I
IIbullt bullt
bullbull Ibullt
1
Inmiddot middotmiddot11middotmiddot f~~ a~ -tmiddot bull J~f
I~ W
i5iImiddot J~a t
ilmiddotmiddotmiddot I middotIt Ii
l
~
middotmiddot~~middotlL= g r
Lj Ic
t
i ~
I
gi II cEi ~ I I
1 t rW
D
tR bull
bull t 10
11
r
i
I ~
j
~
I
~bullbullbullbullbullbullbullbullbullbullbull
LV
FigS pH and p02 profiles into macrofoltling eomlllllllitilS dominated by Balm trenatU$ on steel plate 11 two pH profiles Ib) two lIOai profiles leI pH and P02 profiles into dead barllllCle shiella withdetri~Us I I
itIbl r p02 ~ I
nvnHg 11 70
j ifJi eo 1amp0
laquof it 30 s
I 20 N
ca 11 10 a o -- shy i bull
11
80
19
781 o 2 6 8 10 12 0 2 6 amiddot 10 12 dIpdI Qf ~iftto fouling In HighIst numbIII pIatIi
8deg~~
~i c5 iioo5 fiil
pH 8 bull_ ~
~~ ~
_ bullbullbull- pH
-pOz
(cJ
e-Clf ( j~JJlaquo1- --~I ~a ilil - _ I i ]1
~ ri en i ~ oJt ~iJ id tUI
2u1U I shyI t
shy1 2 3 6 6 7 8 9 10 11 12 i shyDepth 9f electrode (mm) into fouling
1 i
1
~----------~-------
~
~-~ lt1~
middotmiddotPmiddot ~
I
V
~
ill
1 Tbellitt~~ of cathodiC protection OO~~lIic~tlO aridrfOflllUlc
ofi AA~Uoullr14l ~1~I
I
ael~lner EG middotIUtcb8l1AJ Dept lt4 lInY1roMe1lal Bl01011
=~~venilt7 IlariCheW 1f139PL $ ~11al1df
r
I
Ablitrict ~~~ e~l billttI ~ted with ~pert1n ant11_1111ampp paint llere itpoii~1n _vatI- In the tl~ld iUIit 1bOioatbrt~ScM Were
1IUbjll~ to qtbocJ~C ~teltt1on byen _ current (~)SJi inci EDAX Iieamp~ta IJdlctecI ~ lachln teoc copper OIl tbePlCtec~ pa1n~ atter14 dQ1a~ protected Pllnt abo ebowed
) reduCed bacteiial activIty butmiddot ~e4 dltoe Iu1d protOzoanmiddot co~~l~tlont~1l IlU l~byenCOCltact not byen dUfwIl00 ~r _baRl lrillolvedln tOXln nil ereio noted
l ~~
1
I
Introduction
Applications of protective coatings and cathodic protect1on(cp~l
are recognised corrosion control methods for IIIBrine steel Itructureso Cathod~c protectlOll preventl conCIsion by Int~lng til8ctrl~l
currentS trem externalllClUfCa to e~~tthlJno~ e~ carrollon feetlans C04U lUChu tUauJ1ng Aelnts a
(
barrIer to ~ the flao of couos1a1 ~aboIHnu~r
Dccurrlng anodes cathodes or wlwit gelvWc CCIIII1 ( 19681
flinger 1980) AnUfauling ~ also JJICOqIOrII~ b1oc~~
Tr~ylt1n nuor~ (~) end o~ Oxide (~) (~ flJ1J
- of u blacld1111~
V _Ut Evans 1981) ~ ~
end growth of bipfoullng 0I1I)IIIII1IIIIe Wdd~ ca ~ of
paint rusendmiddotmanemiddot ~(r~ til )
As corrosian ~t1on ~ bICCIIIII -an ~1tlQd ~ of
antiroullnQs 1 conjunction _ith CP have bllClIOr ~
resultIng in reseudt 1rwestllj111tlng trw COIIIPIitlbUlty or VU~ ~t t _~
rOratlation to dlfrert levI ot CP (~Eelf 18110 ~
~t 1980) RelatIvely 1nIestiljlllUans have ~ urtw ~
or CP an thIt anUraullngpert~ of palnta
In thIt preseot 1nlIestioaUon tV~iln111 ronutlon ud
cOIIIHrclallY incanjlntlon wIth CP was ~t4IC~ Iw I sraed current (-966 l1li) arid exposed at the 1 Strait IInQlbullbullbully
~t1ons weremiddot 00 the IIhcIrt bnifects the PprIUd_
had an a) the various COIIIPQI1eIIts of n pe1ntf1l11 end b) l1oft~
developaentbull
bull bull ~
~ ~ - f
bull ~ _T~~-middot~ ~~- ~r _ -$1
(1) ~rlttionor Painted Steel Billets middot_ ~~tmiddot_tlJ middot~hmiddot~--~fmiddot ~ middot~A~-~~middot - -~ ~
Thirty-al ateel blllets (8-10aD2 end l depth) were cut frollbullt ~middot~middot~middotmiddot~middottrSmiddott ~jS-~ It bull gt~ tfl - middot--imiddotmiddot ~ t1dItrd steal plates The dimenSlonsof -thB butits-lim chosento
bull -~~ middotmiddotgt2h ~ ~-Imiddot ~~rt ~- -middoti~~ ni ~~middot1 l middot-~t~~middot _
(a) fecilltate amiddot Ie detrucUve -thxI of prePraUonpdor to ~~_gt gt~ k7i ~_ t~ 1--1- _ ) - +--_~ -_~bull ming electron ~ 4II1CI (iialloWdirectobs8iVatlCnS of the
- i lt-- ~ gtbull -o1e billet 8Uface
gt r~middotmiddotj ~L(~middot~middot rmiddot tmiddot J ~ - ~ - lt _ bull Single non-standard (1hIula~) nickel lIIireSmiddot Spot Welded e1Cing
bull ~1deJ ~ b~i~ ~j~~ tri afl~1b1ipOi~ f ~1t1 ~4middotmiddot f-gt -~ ~S-- ( -- - - _bull resin EactI billet then Undb~ tI1d prtiladlllltha5O __ eyilr
bull ~~_ ~~imiddot ~1iil h~ intlfcili~ ~eonbt1nlyi Tr1butylt1n F~id(18n)~~~O~dettu20i~blcicld8is 1IIfii bull i - ~ ( then 1IPI)l1ed to glvaadry Ulil tIi1c1ltnOss or 1(1)-150bullbull
tbull (2) IipreSSeCi Current Syat
bull A rigid ~tic due 1I8S drWedllliihwrtv-s41 liiIihOieS IIPProX~
bullbull 05-10 cia episrt ~ Pilnted ampUl8t Wsfbed IntaPQSIUon ampy
threadlng the nickel IIIlr8throu11 the dinto a tarproot junction
box Eliltilen of th8~1l1ii (recelving ~t-d curnritlere IIIlred
Into bull block connectOrrecelvlng dc trOll is potentiOstat The
rainJng iontroi blUets Wet8fbedfl11i1iiy lIIithln thejtlytlonbox
i~ityd~c IkiliPly ~~ ~lat1ruaWiode 8ncIacalOliel
refer~ electtodetoiere fllied ~ the per1aetat of the plut1cdlc IIIId
connected to the pot8ritlostat The desl~ of the potentlosbt ailoiolEld
~PlDteItlO1TClJrnnt to be MIllntained at 8prampset levtl of 966 II1II
c
r~
Grid raf 1J5s-t 114iH 563724 dud 0 Juna
Alolecule andbechr1a in general are ~ to
sutface lI1eroleyer (nlUSton) of _ter (~bullbull 191KJ)~To avold
contaJnatlonof ttw painted surf by U- -vents
blllets Were s-nact in a 1 conte1ner or atc1le (02i ) f~~nd
ter Iih1ch then f9ICI in tv 1IIhere the b~
be r-oved undeNeter El~~ bllleta fa Mcb trtai bullbullel
removed over a 1 dtq (336 hr~ expoeure perioda
each being ~~bn uposurnof 1 5 11 iA 168and DlIIauz1r
Arter field axpauHli the billets wan
fUtered _tar to ~ 1_ debtlflMd far 10 aJnuta ~
wv ~~~endattid sdbUiUd(otO
~ ~l f~ nn1ng el~~ --
lOOted onto 1middotCIi ~bullbullWIsend carbonmiddotcoa~nmiddotmiddot~middot f
each billet Ws~ Iis1ng a ccr1dge ~el~~
(5360) IIIlth anergy dJ8perslve X-ray alcroenelre1a (mx)
_lysis the les Wen reNed end coated III1th vold to al]Ql En
detailed sat of the alaquofa1l
(4) Lfiborattux ~
Painted taa~ bUletalllllre pnparad ~
in a Mrlne reclrculet1ng flOlll _ ~
f1Oll the ~ ~a1t~y Tha~~Un bullybullbullbull
10C M1nta1ned ~~t1as of ~t~la protazOllbull~bull10shy
(Entermap) ~ ~~~middotNlt~Eamp1~ __~and ~ r - ~ -
certaJn ~lc 1IQ1~ 1(1 the
the painted
~
thre repl1catft fn
plKed 1nto(022_)
- thIine1J~~ -lt bull
Atter mAX
(1-2) and i-n-t bn collected
14OC
Pain~
-
12 daY~ exposure and prepared as in d)~ - t
~I ~~~-- i~yen 1~ v
Rflltsi ~ _ ~f-~lI ft j bull _
rt~ll0~~~llY~~~~peln~stetsl~11i~ex~sed for
y~t1~J~ ~~~~1~1~~1UII~ya~kooJ~~~lt ~ ~~_~~I1J~ ~~~tomAX -lysis ~era than gold ~ bull bullbullbull c - _ bull lt
~~fllr ~~~ vllUll ~t1ans of the rfee htures and gt Co
~f~ - ~~~ ~f ~ ~aobta1Mdue IjIll811 iI1Flg 1~2 The IIIIdn ~~ ~ lt)ji9 )tmiddotmiddot - ht
- bull
~I rIIplt~ tIt8 -~ cOlllponanta of the pei1t 1bullbull WelIIlItr1x $ilj~t-~~middot - ~ _ -_
-IUch PVCbQed end rap~13y the chlorlna peak the
i~~~-~~-~V~~~-t18 ~~~~ ~~ Th8
ral~~i~ JlrQP(lrUo1sof ch 1Il-ntal paek~ll1d l~t_ Q~7 -gt- ~ _~ -- gt ) gt -_ -lt ~~ the 0 -end _ xprtsntabullTIwsa changes are -r1sed ~t~ lt~ _ ~ 1nT~ IThe8llia$lb npresentthe chengelI 1n~t1V8 p~lons
ti~~~ [ ~gt 3 - - - _ - _ -
Qf nellllllllt within We surface layars of the pe1ntmiddot ru The
~f~~thtJn -d ~rfeli avr the 336 h11cxI ampScanbe
SII4iIl r~ fig 3 al~ tht rta of~ dirfers tJetwaeo thaa ltmiddot~1- 1ft~- ~ _
IJOl appfiai-ed to ~1nlt1elly until ftar day 168 it fall
~~l~ 1h8relllllrelgn1fi~ diff~be~ qrOtactlid and
proieCted (at -966 ) piIIllnts tor lioo COPPer but not folimiddottin
Ibi~tlons Of the Pain -urface 8hoWad _Pil ~ witilt1M
(Plete I to IV)~ 1hePllat1na (~) pelnt(Phta x and b)
Clearl~st-i~~r alots on the SurfllC8 IiII1kh~ter1sed ~- f hlt1 lriitUl tin conCantrat1on~ lllariy slotsre Partially
~OIIe~ ~ tile _tr~tedal middot1lere rs ~an~ densely packed
grerltr~ ~th 1ar~(iIa1niy cotifIIr) 8ntt -ampniBil (lIB1rily 1rori)being
middot vl~l~~~~~ Dtt~i~~~t~ r~ ~f~~ I~t seawatllr for 11 n IIIOre slots appeared and th~ ~~ess _trb cwer
--if -- V~ c~~ f(i ~~~i~
(Plate Ie to It)~ the granules dld not protrude from the~rtace in ~ ~~~~j1~r~-ffJmiddot0~~
suhap~~r Both 0 nd --960 at ~~pa1nts ~ - fr ~~~~-w1-U~ ~~t
sl~llrat thi stag~ (Plates Ic-d II1d xrnspac~IlYh middotmiddotWith middot middotaJ~cmiddot gt incraallO ~alolre t~ difrrqcbullbull middotbatIiI8IImiddot the protected fjd
bull ~ ~ ~t~
IIllrotacbldpaint ~~lIfIt~ftitr~ h alQt~ p~t bull u bullbullbull J ~~i~~ 1~~
lIIith IIIuper lIdguon u 4lrotatad piIJnt (Pl8ti1 II) 1hD1II8S
Ill )1IMdw1lillS gnnJla~on f-r~~ dpert1cl ~ - - - ~~r ~t
Onthe 966 1IIpetnt t-var tIIO typeaofan with diffannt c t
~hIId~(Plate UI a to c) One hIId tIII-1 bull bullbull e _ bull
fUll wJ1hleas abvlaus alats the o1fwl bed C18UU1IlOt ~litUi middotC c ~ ~
less ~ t11 h Gnnulatlan __ but ~lIiIZly def1nlld in
middot~ latter Afte fila bed tiIIrtIId 10 ~are ~~j(
extansll(tlly ~ the~tIId(O at) tIIIIple lIIIdngthe~ - ~
middot orarallet1ai leas claaI (Plate I~) f u II1ao CQ1tJAJ11d to ~Gn - -
the -966 at 1 (Plata III d tor but 1JI8V8n 1nittNdcneabull -
Althatqa -middotslots wrs abvlaus becaI1ng 1 cwampr aiI- the orwWat1an8
f1nailyftar 14 day exposure ~ fUll drMIlapIIIent on all la - ~
are extensive with the 0 lhawlng both alota pore ~
OVIKgrOloln (Plata II c to f) Gnrulatlon 1bl to ~ gt~t~~ i
or the th1cknats of tho ma ~the ~ at ~ (~~~~ themiddot - - _ middott ~ ~ ~ - -1 1 t
middot f11bull IIIIIS IIOre extans1 thIIIn at dIlya but IIIIIIIY bull1Q~ tI st11l ~w -middotr~ 1gt ~1 ~
obvious fIIany ~ loI8re still p~ yen~~~~~~~r
than before Poiu appearedto be f~lf~ila~~c-~~~a (Plate IVf~ label 1) IIIIl1ch broke~i(2)~ ~~~~
t t
bull ~lS (3) ~ rflC~~ ~i~t~~l~~KI~ t~ldiIiiicyul~loIiih ~~Rht ~ )~- -~ - ~middotd 40 ( gt ~gt ~d lt1 t ~ Jt ~remiddot4amp tha paint trix cavity bottom eroded away reVe81~ng ore
t ~~t~ por~~ ~l~ P~]Y p~~8dby di~lutiaA~ ~~~~ti~Jr~gt~ta(l~~Siinll1cl1Ud~f iUiIiI reriatlGllbullt
bull a~~ ba~tid colCriAtloriccic~ GIl ~~~d ri -966 - ib~~~amp catQn1iti~~t~I~~(~t an ( ~
t the ~~~palnts (PIa_taiiI d~~h ~wh ~ n td have ~ CamplluJMitiIVc) ~~middottti o~lgtnalrf tlt of hIght
~ l=~A ~J~rW tYtles~ p~fiiilriQcOCCold to~Ja to jU-t ~jmiddoti~ai dlatClilS ~ir -jlati ~tbullbullent aNit ~ t ~~ th8middotf~ld~ies~~t~on~ ~ -966 prQ~~~(P14ta Iii~)Omiddotiliof~m1hiiHAiiipieat ere~ll-(piitir~ p-iot~iilans(PiA-iII~ end c) nbullbulltiledt dl~by1i~~ end wirinali1sheii ~cMI~bY thI tili
bull In t-~~ ~1i1nt 1oIIIeHfd~ ~m1IS (15OClind a t ri~r-dlatoa ~lor P~t dlataiOlonlt1anGIl ttw-966
t
bull ~
pr~t~ ~is~ih iion obitJauIJ A ~rot -=1 ~ ~ i __
reprasenttild on the 966 painted 1iUrfc1ncluicunljj ~ rwt
Achnanths Colonisation by bacteria 1n the laboretory bsed
bull eliperlMnt tollMct sbiUar pattern to the fild ~
bull bull Dilicueii
bull bull DUt 1Iidnt ~lona Mve dlttllrint ~~ ictian
It ~ JntM l1tenture ttt Iff yen __ JIll the
bull ~ bicins ani 111 ~t INchlnil (PhillIPs 1t73t EWene~ 1981
In ci1ntlHt tJWY ~tit ~Wl1c iIuCh as 1rbiltyl tirI
~ lite wAlII]ltiClr dlsP8tfu~ middotthiirsttilll~intaJn~ a
bull ~
lIIifoflll ampalCentraUon through the filII 10 thoIt biocide rel_ at tha
paint surfce 1fI$Jlts in IIIOre ditfusing ~s to repla~lj -iflJ~it
resulting 111 bullbulltNdy lr leechlng rata M1llat this IIIIY be middottruIi ot
tin ~ it was nOt true ~ ttw middotlBlf fOZWJlatiGn ua ~ bull - i - bull 1(1
ObSfvt1onundr SE clarly ~ tM t1l1 to be present
~ly lIS OIIIdle-llk parUc1~~t tn lUlfiice (Ito
IV) bUt usa ~ the dIpth attt fU-middot ~y~
If one considra the concn~rtlonmiddot chMge 111 1n ~111c COKICIl8rlts fro- the paint tU- i bullbullbull tMWei b~ ~ Si6t arid
tI-~t iron oxidt 1~ can bt (Fl_ 3 IrId TIibl8I) ~t ~ ase cona1derllble d1fftrenC8s ~ a Un decz bcdYMr ttw p1iod IIIIwraS lron~- an unt Ught inCr h ~ incqJper ~tlGn occUrnd _idly at ttw ~~ Gfb ~ period ~ out ttwn fu nPldly ampQII1n ~ umiddotirid II phast This IIIOUld bt tIlq)8Ctad It cantact l-=h1ng we occun1ng
IIltfIOS8d pertlcl of copper would lMch rapldly on t-n1on 1n - shy JIsewtr There would thlll1 ba bull J-a a1iltUy leu avalUba
i1 ~ ~
_tedal becfM uposed Thi tIOUld ttwn IIllCh Thla pulSing efttct
IIIOUld be expctad to -oth OIItlllith U- putic1 __ -lllible II tot 1~ GIl a on r-x _lao Thla teet 01 put1c1e 1tUng
can tit GIl the lIUrlace ClIlC8 it ~ a pan 1amp t
a 11aM bet in Plat I to IV IIaIrIg _ lit la_ II ~ 0- ~s _- PIate IW $llldl ~bull nshyot poRI flllNltttn Al~ tin ~ lou rete thtn I bullbullbull atill Ivwnce of bull al~t pulsed 1Itue rrlict tII1IIWd
elqlCtaG inIl ttwauUr ~ I I I
0
~ i
~ ~
~I J
bullbullt bullbullbullbullt bull bull ~
bull bull
I
bull t
~~
The same seqiHllle of cevity follliltloii (intttecaseofthetestjjltle
slot- shaped) fClliOileciby SecOlld8rYtlolesfotlllingin thll cavity
bottollis releasing further deep8rtox1ils (Plate XV d and g) As
cavities_re ProdICed ~ tiecaaIelnvadedby_ter~ onelilauld
expect 1ncreaa 1n1IOdha and dIladne lMls t the t1met
which the toxlM uw ~ Ths fCMld tCtHi eo (Flvmiddot 3)
~rttno the vi that_tat ~t1on otthe~ ~lY
il1Cl1l8S8S Idth UIiJ
Differences f~in toxin li~~to Ini 966 ill 8811P~ F ~ bOth Un end ~ lcMIr levels afillW
IIIIIlfI ptaInt(~1~~1ntha~~ ~~f~~ cst rklld with tbiIccpper ElJIc~l ta~Uona invalvinv
w pl8cId at a~ht~~middot3lO~lth Un cqJpel bull
(-960)10 ttW ~af this expei1tIIri1ts thIsiorelfJltdt -
that cqlp8r b8IiIId ~ IIiIOUldProc-ln f41ip1dly t thellllgllitiv
patentW _lied () as 1t 11 further f~1t ~ witt Un
III11ch is u-tat aqulllbd Idth
it If ttda 18 eo~ ~
~ )
frOll1 paintscould bl anqnced by an illprd current clithodic
protecUansysteipos1bly ahart_nino te 11fe of the paInts
antifouling ability Bacterial coloni Uon IIIU rtdUCed on the -s66 paint This
could have been due to unflSllOUlllble electrostatic cqrge on th surtece Th1a effect CIIl pley en IIIportant role in the t~of
bacteria to alrraces (lIIarsMll19E1O) As bacteria appear to IhiJw
greater toxicltyto copper (DMpsey 1981) it is Klre likely that
enhanced cqJper leacll1nl On the -966 AIIIPl c8used the recluCed
activity The IDlol8r toxicity ofUn to bacteria (as reported by
~ -
DelIIPsey Hle1gt or its lowes leaching rat was sUpPorted bythl
relatively rapid recolonisation of the slots by lIIicroorganiSlllS ~ftar thE inlt1al aurge of leachi~ Thailllpressecl current dto ~~ the oppoSite _ff~middot ~r on dtta colonstJon l1lI-1- in the
JIbcJrtory~ta larger rUDIIra f~ fit pro~td ~ther than uriprotected Pafnta
Ccnlus1one
1 tiild-a 18 presaIIUd ~ thattJn~ ~~
~llNChng notd1rf~ ~
2 TIleuseofCP~ ~CUIftnt(at) on1h1a ~fca41ng ~- ~
pa1I]t~ to ~_lAIIach1ngn_ at toxiN - -
3 leIJstaPidbectar1al ca1on1aatJon acaampmId an ~bId Pt1ntJ ~
4 JItore larger alcrofauJing (Oiat_ and PzOtoaoana) Gr~_
occurred on the protcbld~
I l
I
bull bull ~
mfEflpoundaS middot~-t9i _jgt- ~ _ _ ~sev M~J~ (1981) (cilonJzatiooof alUf~lng lnts by _rine
Jbull ~teda9g~1ta IIarina XXlV~ 185-191
- - gt 0 Ie 1 _ - _
poundV8I$i I~lf (1~P~h 1II8rin~Ugae and Foul lng AReviw ith
t p~l-i9Jrlr ~iiMdilto5hiP-fQUllfCiH 8ot~ ~r~ ~~~ XXIVbull lIPwl~J11A
Fotteth CN~R ~kk8nGB IWPIR1seJCiiiroaic1lihd ~e ~JIIth CII Offshore St1ucturu ElllS ~tjI~f~1Dt19i4
- - ~
UKC (1976) I~races ill ~W Ecology KIuverd Univbullbull middotraabullf~iddge ~bull C~~jII illt)Ia(t RC (1980) totinllsa Cattod1CPrOtect1cinbull middotmiddot8AtEmiddot~6haOCIlfif~ T~ p No 108 ~ 18 ~ 1IIHf bull
t bull lt ~J~~~ __ ~
Norkr4irulB(198D) Advanculn UcrOfj~lpound~ltiOyVol 4 (AlItxlaquoIder fl Ed) PlerIw Press lcxIdon~ lIP~ 51-85 ~ l~l - bull bull
PJijllt~i At(fm)~mr~~JnI~1fqul1nQ JlalntlJ Riumh ~JnOrpnic toatJngs 2 192middot
)~J gt0 c ~lt -- ~ __ RogerS~ Ut (1968) ina ~(lnt~fbItliP4ili GI COnoslon ~a Technology)bull bull
Sl~iorr~ VP~Ranald~ Rctd~~ (1S80)_ ~ Experllattal Stud1aiRelate middotEffect or Cathod1c Protict1Oi withmiddot terta1nr1c
Coating Syst~ 12th Annualofr T8Id1na1ogy Co1fItHnC8 liauston TIIlt p 9-12
j
fIG1 sPct FromMon IT8TJCuZOI Int Lliu EOAX
FlG~ Fro TlTCu20Ar-4 Dy Field Ellposult I~ir
il
I c
R hi -00amp=7
-I I I I I I I I I f
iii
bull bull
bullbullbull
I J
J
I Figur 3 Thl chan In reltl rqpoIQnlof -ariOtimiddotjOmponents of the paint film with time Thl vtic~1 ampkiS unl till _n ofmiddotmt Illment per unit OJ fe 50all Cl Na
Cu StI Feeshy
PlistinllFe S7~J6 704 3~74 908
f
r e ~ ee~
middotie ~lt - ~ shybullt bull ~
t -~ l Cu~ o ~ f
t --- 3igt - bull 0 J Sn
cc
t ~
~
bullo
~ bull e
bullbullbullbull bull ~
bull ebull a QI 1- bull 0 40 431282 316
101 dmllIII)bullt bullbullbullt
hr 0 ~~bulla 5~16 ~~ -~~ i166 4S i83 8~1B
24tu 6~480 SD~83
111middot 1~1 OS6
~ 5114 6~69 -51 UIamp middot016- lt ~rmiddot
1d o Mea 818 21 021
~Z21 199 1~1Qmiddot ~Jgt2 058
14 d o ~~ 804 Q~B8~~~a~~t~middot 685
866 372amp 8amp4 OS _i~11[ ~~64
Tttile 1 A81UvlL pzQJJortlcw 0 vxl~oI ~~bull Jn 1I1Dtitctel lt-1166) end lqIlDteCt6 (O)middot~af~_~middot_lodEDf~ middotmiddotmiddotmiddotmiddotmiddotmiddotmiddotmiddotc
~ ~
_----I~
J
I bull
I
J bull tmiddot
7o CONQRESOIMTERNACIONAL
I D~CORRO$IOr(IAAINAEmiddot~USTACIONES~v c)
t 7thflfERHATlONAL CONGRESS
ON IARINE CORROSION AND fO(JUNG
~CONGRE$INtERNATIONAL DE~CORR~ON ~E ET QES~SAUSSURE$
i t
~ ~
bullt
t t
UNIVERSIDAD PourtcNlCAVollncio7J1 NiwiemIif~ 1988
(SPAN
SECcl6lfn
BiOIogro marino Morine biologY ~1i1orine
Theunpub1ishedJllBIlu8criptEl have been reprod~d asr~ielved from the
authors The SCientiUc COlIIIBittee take not respoi~ulbll~~Y for any shy
error or omssi9n
Laa ponenciasno han side publicadas y estanreproducidas tal y como
sellan recibido demiddot BUSmiddot autorea pound1 Conte Cientfico middotno se responsemiddot
bil1za de cualquier error ~ om1si~n
bullbull _ bullbull_ bullbull ___ bull $_ _ L _ - _ --- -__
SESSION II Marine biology
Friday 11th November
SURFACE PROPERTIES ON NONmiddotTOXIC ANTIFOULING PAINT FILM
K Nanishi M Murase Y Yonehara M Kishihara and T Hirama
MICROFOULING ON ALUMINIUM PANELS PLACED IN THE TROPICAL WATERS
N B Bhosle K Venkat and A B Wagh
EFFECT OF TRIBUTYLTIN OXIDE (TBTO) ON THE OVARIAN MATURATION OF THE PRAWN CARIDINA WEBERI
R Nagabhushanam B Indira R Sarojini and Mary Frances Thompson
EFFECT OF BIOACTIVE MATERIALS FROM SOFT CORALS ON BACTERIA ASSOCIATED WITH COMMON FOULER
Vitalina Mary Avelin Mary R Sarojini and R Nagabhushanam
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
11
10
the depOSlts woUld terid to Ile lOW at We start of ~r1ments as
condit1onswere nat favourable (POstgate 1979) The cantlooQUs
prockIctiPn of r1th f~plieUdofeacal_te1alor IlUlpllonill1l
~ts bYpoundilt8~ (~~Iick1ngand 1IkIod1) hOweva ~ t H 1 (
IIIOUld rep1dly altAIr theli11l~t and the rua~ofSRB re
notedtofncreasemiddotttcllll ( 1 SRaIll1taat llaUt 1 x 101 SMIIl-1
aftet bull -th
IUctotJlIIl corrosion canfe=t _ta1sln a1Ullberof Ways
(Iverson1972 fUlllIt end King 1975f 1Uilr 1981) pH can
aff~ the ectivity of the 1IIin organlasinuolved the H 1IIho
hava tin qlu pH range beban 10 12 (Graff 1981)
although tIdS -can be lnf1u8nCed by ather fllltors In atI1tl11n
they lIKIJlre a catbon end nitrogen aurCa 11 various
salts tin alactron accaptor (HedJart ~ idbullbull 1985)bullAil of
U- facton are availabll in the debris -alltting b41tween
ar1na foullng -d the 1lUbstra~ 110 providing bull potantlally
corrosiveanvirorwent
Evan IIIhete such accullIUlatiori does not take place varlable
changas inoxYQ8ll and pH ~trations acrossthe suface of
teel soch as C3I becaustd by foul~ ~ C3I give ti_
to corms1an calla lalildlng to piftlng corrosion
There is an initial tllld8llCY for the pH to rise as a relUlt of
the excretoryactivi1i~ of fouling aniIIal and this 1amp UIIllkely
to give rise to corros1an problems Eventually ~er feacal
h
1
~I
materials and detrltus~ill build up around or~anisms ~re bo~
low pH and p02 devul9ls providing a potentially coJJOampiIu
envinx-nt flaw does dify thh _build UfJ but inmiddotthbull
Ixparlunts carrllldotttid notprlvl(lt it R~~r it
l8Iilated the depth of deposit ~1~~ in t- ~ It ahoutd alao r~~- in f~ population so-1QIJvlWela 111111 be dying or dUd Thlb
~lng~wUl~1Ielacal1Md conoa1ve ~iccandltiorJs-t SR8a 111111 b4I fCllllfC314lng ~y pOIi1desa hlbitt~ HtInd 1tt~ WIi~ul create 1~1Mct Jrcx-ntal dlffarenca ~ ~th l1kel1haOd or pitting
n
References
Albilnder J~T 1 bL~1953~~(9EthII prockIctiPn of IdcroaltJodIIs UIIId in I~ studies Rev Sc1 lnstra ~ 528-531
_r1can Ptbl1c Health laSoc1ation 1975 5taIIdIrd~for the lIIIIIlystS of _tars end taatms 11lth pounddIt1on 1IPHIi-~ 1015 ~taenthSt NW 1IIa-l~~OC 2IJ]J6 USA
Bau-8laquo=k1ng C~IIIG amp 1IIoOd EJF1955 Biologlcal ~ in theestuarlna envi~t 1 EcolClil) of the ~ cycle Kcnlridlkjl Ned Akad -t ~ Ser B~ fi 160-181
cter DB amp SUver IA 1961 In IRafaranctlElectrodl$l EdltolS Iva DlG and l-z Gl lc8dilll1c Pr London Maw Yodlt Chapter 11 ~
Davies PIII I 8r1ric middotF 1942JII1c1oel4iitrodas for-aln local oxygen tension in n11 tissues Rlv Sci InStruII Yo 524-533
EdyileariRJGJ ~ Tetty LA UJ83 The 1nfluenc ~lIlcro algae on corrosion ofstructural steels used 4l~ the 1tIirth
Sea 81odeterioratian sect 336-347
bull bull
~
bull - -- ~It Jf bull
- ~ 13
12
Terry LA I~Ed)VeangtRfiJ 1981 lIi(toal~IOI1CGrmianF~ l)i 1St1oJhemailnEifooiiiigffixed gftshoreIt Bot 1Iar ~ 117-183 lsect~~jlaUoQ1l1 _ ~t Ef)e~gy Offs~r1I Te~lOtjjy Paj)8r
-middotltcntpI)~it$t~l91-16PP~middotmiddotdl ( I
bullbull
iIIalk e1 JL 1971 bull Ion specfie Uquid I~~ e)(charig~ ndcroelactroclels Anal ChIIaI ~8S-Q2 AGraf1~ w~S 9S1~ 1orrosloh InFIntroooctI00 tooffshOle
51~weSoes~~fab~lcat~onIns~allat1on Gulf Wall PI aelUnger EG 1982 The appl1~tlon 9fMl1ikIngmiddotHoust(jfW~Q23g-258middot icroelectrade in the study of pH end PD2 in lIHinli
HerbierC SN fGH~rt~ )RI -Stoctcdalpii Watltif1Sal RS ~1ronIIInb Ub~~ 27l-Z79 1985 Fac~or lontro1l11)9 tha activity ofmiddot sulpMte
bull Wall P 1983 The appllcatlon Md ~ ofiwrJ)e~c~odl shy-taicfii9 tiiCtetili tn~ diiilncj tar infection
techniques for the iiaurnt cif icraeov1ror1liltital5QiCiety ofJebo1- Engi~ SPE 139781-10 _i~~ - ~ ~ i ( _ IKIdif1catlons caued by aGII8 rlnefou~lng -C~ebullbull
PhD lhaaia Ibiverslty or ~tar UKbull bullbull bull -
IIH1nke JA 19fiSa In EXperllRants in Physiology and middotampioClIiIiiilatJift l c E(If(orltGAiKetkutgt ~c-Presa-
Wooltdngton AObullbull Davenport J 1983 pH lind IlIl2 IMla ffl~- j~t~5 -lMIIleath 1IH1na ro-fauUngGqWl J bull Expbull 1
B101 Earlsectsect 113-124 Klnke JAIII 1969b II) Glass IUcroelectJQdes poundx1tors Uwal1~Ui~ OrOllt arid~t Nt JOhn fII11ey YOI1lt thaptar 11
~-f _f~ ~ ~~nt ~ Igt
bull Iverson IIIP 1972 Biolog~ Qrroaian In Advencesin
corrosion scianca and tecIYIology 2EdS FCntaia G~ I bull IIS~et RIIIbull p~ PJaSsLancIoIl 1-42
bull
Jorgensen BB Reusbech NP I Cohen T 19B3 Photosynthesis and strUcture of bIIn~ II1crobWp MC1OIIlectrocki and ~ stud1ea of four cyanabacledalt -~usllWtOl ~bullbull 1~f093
t l~ O~ ~~~ H~ ~9s7~c-gtHatste~iecfn1k van shy pellediftlrtenPt - Stlchelektraden (1-Su
AiJBitiKlificfiliilislflilr)zlIr polrapusdiailJIIissui1g_ liIserstoff(jruQkas fur d1eB lIsUbullbullung der IIliJlttOUf1cillatlon PflftgaHArdI ~fl39
bullbull Mller JOA 1981 tals In EconcinI1c II1croblolooy sect McNbUdbicldatariOtationEd Rosebull A middotAcadelc Praulondcn149-202
t Mller J DAI King RA 1975 8io$~iorat1on of lllltals In lIIi~1al ClflPlCts of the deterloration of tarWbullbull EdsGilbertRJ~ middotloveloCk 0111 bullAcadeic- Presst IILandon 8Jo103
bullt
IPotgate JR 1976 The sulphate-reducingbacteda
Callbridge tIn1versity Press Cliilbrldg8U-K
Reusbecb fUI i Ward DIII 1964 i~~~1Ida5tud1el of
bullt inter$Utilll IIIItez dttm1stry and photosynthetk activity
in ilflaI spring Idcrob1altAppl~EIi~ MerobW tsect4IlU-27lt I
IIbullt bullt
bullbull Ibullt
1
Inmiddot middotmiddot11middotmiddot f~~ a~ -tmiddot bull J~f
I~ W
i5iImiddot J~a t
ilmiddotmiddotmiddot I middotIt Ii
l
~
middotmiddot~~middotlL= g r
Lj Ic
t
i ~
I
gi II cEi ~ I I
1 t rW
D
tR bull
bull t 10
11
r
i
I ~
j
~
I
~bullbullbullbullbullbullbullbullbullbullbull
LV
FigS pH and p02 profiles into macrofoltling eomlllllllitilS dominated by Balm trenatU$ on steel plate 11 two pH profiles Ib) two lIOai profiles leI pH and P02 profiles into dead barllllCle shiella withdetri~Us I I
itIbl r p02 ~ I
nvnHg 11 70
j ifJi eo 1amp0
laquof it 30 s
I 20 N
ca 11 10 a o -- shy i bull
11
80
19
781 o 2 6 8 10 12 0 2 6 amiddot 10 12 dIpdI Qf ~iftto fouling In HighIst numbIII pIatIi
8deg~~
~i c5 iioo5 fiil
pH 8 bull_ ~
~~ ~
_ bullbullbull- pH
-pOz
(cJ
e-Clf ( j~JJlaquo1- --~I ~a ilil - _ I i ]1
~ ri en i ~ oJt ~iJ id tUI
2u1U I shyI t
shy1 2 3 6 6 7 8 9 10 11 12 i shyDepth 9f electrode (mm) into fouling
1 i
1
~----------~-------
~
~-~ lt1~
middotmiddotPmiddot ~
I
V
~
ill
1 Tbellitt~~ of cathodiC protection OO~~lIic~tlO aridrfOflllUlc
ofi AA~Uoullr14l ~1~I
I
ael~lner EG middotIUtcb8l1AJ Dept lt4 lInY1roMe1lal Bl01011
=~~venilt7 IlariCheW 1f139PL $ ~11al1df
r
I
Ablitrict ~~~ e~l billttI ~ted with ~pert1n ant11_1111ampp paint llere itpoii~1n _vatI- In the tl~ld iUIit 1bOioatbrt~ScM Were
1IUbjll~ to qtbocJ~C ~teltt1on byen _ current (~)SJi inci EDAX Iieamp~ta IJdlctecI ~ lachln teoc copper OIl tbePlCtec~ pa1n~ atter14 dQ1a~ protected Pllnt abo ebowed
) reduCed bacteiial activIty butmiddot ~e4 dltoe Iu1d protOzoanmiddot co~~l~tlont~1l IlU l~byenCOCltact not byen dUfwIl00 ~r _baRl lrillolvedln tOXln nil ereio noted
l ~~
1
I
Introduction
Applications of protective coatings and cathodic protect1on(cp~l
are recognised corrosion control methods for IIIBrine steel Itructureso Cathod~c protectlOll preventl conCIsion by Int~lng til8ctrl~l
currentS trem externalllClUfCa to e~~tthlJno~ e~ carrollon feetlans C04U lUChu tUauJ1ng Aelnts a
(
barrIer to ~ the flao of couos1a1 ~aboIHnu~r
Dccurrlng anodes cathodes or wlwit gelvWc CCIIII1 ( 19681
flinger 1980) AnUfauling ~ also JJICOqIOrII~ b1oc~~
Tr~ylt1n nuor~ (~) end o~ Oxide (~) (~ flJ1J
- of u blacld1111~
V _Ut Evans 1981) ~ ~
end growth of bipfoullng 0I1I)IIIII1IIIIe Wdd~ ca ~ of
paint rusendmiddotmanemiddot ~(r~ til )
As corrosian ~t1on ~ bICCIIIII -an ~1tlQd ~ of
antiroullnQs 1 conjunction _ith CP have bllClIOr ~
resultIng in reseudt 1rwestllj111tlng trw COIIIPIitlbUlty or VU~ ~t t _~
rOratlation to dlfrert levI ot CP (~Eelf 18110 ~
~t 1980) RelatIvely 1nIestiljlllUans have ~ urtw ~
or CP an thIt anUraullngpert~ of palnta
In thIt preseot 1nlIestioaUon tV~iln111 ronutlon ud
cOIIIHrclallY incanjlntlon wIth CP was ~t4IC~ Iw I sraed current (-966 l1li) arid exposed at the 1 Strait IInQlbullbullbully
~t1ons weremiddot 00 the IIhcIrt bnifects the PprIUd_
had an a) the various COIIIPQI1eIIts of n pe1ntf1l11 end b) l1oft~
developaentbull
bull bull ~
~ ~ - f
bull ~ _T~~-middot~ ~~- ~r _ -$1
(1) ~rlttionor Painted Steel Billets middot_ ~~tmiddot_tlJ middot~hmiddot~--~fmiddot ~ middot~A~-~~middot - -~ ~
Thirty-al ateel blllets (8-10aD2 end l depth) were cut frollbullt ~middot~middot~middotmiddot~middottrSmiddott ~jS-~ It bull gt~ tfl - middot--imiddotmiddot ~ t1dItrd steal plates The dimenSlonsof -thB butits-lim chosento
bull -~~ middotmiddotgt2h ~ ~-Imiddot ~~rt ~- -middoti~~ ni ~~middot1 l middot-~t~~middot _
(a) fecilltate amiddot Ie detrucUve -thxI of prePraUonpdor to ~~_gt gt~ k7i ~_ t~ 1--1- _ ) - +--_~ -_~bull ming electron ~ 4II1CI (iialloWdirectobs8iVatlCnS of the
- i lt-- ~ gtbull -o1e billet 8Uface
gt r~middotmiddotj ~L(~middot~middot rmiddot tmiddot J ~ - ~ - lt _ bull Single non-standard (1hIula~) nickel lIIireSmiddot Spot Welded e1Cing
bull ~1deJ ~ b~i~ ~j~~ tri afl~1b1ipOi~ f ~1t1 ~4middotmiddot f-gt -~ ~S-- ( -- - - _bull resin EactI billet then Undb~ tI1d prtiladlllltha5O __ eyilr
bull ~~_ ~~imiddot ~1iil h~ intlfcili~ ~eonbt1nlyi Tr1butylt1n F~id(18n)~~~O~dettu20i~blcicld8is 1IIfii bull i - ~ ( then 1IPI)l1ed to glvaadry Ulil tIi1c1ltnOss or 1(1)-150bullbull
tbull (2) IipreSSeCi Current Syat
bull A rigid ~tic due 1I8S drWedllliihwrtv-s41 liiIihOieS IIPProX~
bullbull 05-10 cia episrt ~ Pilnted ampUl8t Wsfbed IntaPQSIUon ampy
threadlng the nickel IIIlr8throu11 the dinto a tarproot junction
box Eliltilen of th8~1l1ii (recelving ~t-d curnritlere IIIlred
Into bull block connectOrrecelvlng dc trOll is potentiOstat The
rainJng iontroi blUets Wet8fbedfl11i1iiy lIIithln thejtlytlonbox
i~ityd~c IkiliPly ~~ ~lat1ruaWiode 8ncIacalOliel
refer~ electtodetoiere fllied ~ the per1aetat of the plut1cdlc IIIId
connected to the pot8ritlostat The desl~ of the potentlosbt ailoiolEld
~PlDteItlO1TClJrnnt to be MIllntained at 8prampset levtl of 966 II1II
c
r~
Grid raf 1J5s-t 114iH 563724 dud 0 Juna
Alolecule andbechr1a in general are ~ to
sutface lI1eroleyer (nlUSton) of _ter (~bullbull 191KJ)~To avold
contaJnatlonof ttw painted surf by U- -vents
blllets Were s-nact in a 1 conte1ner or atc1le (02i ) f~~nd
ter Iih1ch then f9ICI in tv 1IIhere the b~
be r-oved undeNeter El~~ bllleta fa Mcb trtai bullbullel
removed over a 1 dtq (336 hr~ expoeure perioda
each being ~~bn uposurnof 1 5 11 iA 168and DlIIauz1r
Arter field axpauHli the billets wan
fUtered _tar to ~ 1_ debtlflMd far 10 aJnuta ~
wv ~~~endattid sdbUiUd(otO
~ ~l f~ nn1ng el~~ --
lOOted onto 1middotCIi ~bullbullWIsend carbonmiddotcoa~nmiddotmiddot~middot f
each billet Ws~ Iis1ng a ccr1dge ~el~~
(5360) IIIlth anergy dJ8perslve X-ray alcroenelre1a (mx)
_lysis the les Wen reNed end coated III1th vold to al]Ql En
detailed sat of the alaquofa1l
(4) Lfiborattux ~
Painted taa~ bUletalllllre pnparad ~
in a Mrlne reclrculet1ng flOlll _ ~
f1Oll the ~ ~a1t~y Tha~~Un bullybullbullbull
10C M1nta1ned ~~t1as of ~t~la protazOllbull~bull10shy
(Entermap) ~ ~~~middotNlt~Eamp1~ __~and ~ r - ~ -
certaJn ~lc 1IQ1~ 1(1 the
the painted
~
thre repl1catft fn
plKed 1nto(022_)
- thIine1J~~ -lt bull
Atter mAX
(1-2) and i-n-t bn collected
14OC
Pain~
-
12 daY~ exposure and prepared as in d)~ - t
~I ~~~-- i~yen 1~ v
Rflltsi ~ _ ~f-~lI ft j bull _
rt~ll0~~~llY~~~~peln~stetsl~11i~ex~sed for
y~t1~J~ ~~~~1~1~~1UII~ya~kooJ~~~lt ~ ~~_~~I1J~ ~~~tomAX -lysis ~era than gold ~ bull bullbullbull c - _ bull lt
~~fllr ~~~ vllUll ~t1ans of the rfee htures and gt Co
~f~ - ~~~ ~f ~ ~aobta1Mdue IjIll811 iI1Flg 1~2 The IIIIdn ~~ ~ lt)ji9 )tmiddotmiddot - ht
- bull
~I rIIplt~ tIt8 -~ cOlllponanta of the pei1t 1bullbull WelIIlItr1x $ilj~t-~~middot - ~ _ -_
-IUch PVCbQed end rap~13y the chlorlna peak the
i~~~-~~-~V~~~-t18 ~~~~ ~~ Th8
ral~~i~ JlrQP(lrUo1sof ch 1Il-ntal paek~ll1d l~t_ Q~7 -gt- ~ _~ -- gt ) gt -_ -lt ~~ the 0 -end _ xprtsntabullTIwsa changes are -r1sed ~t~ lt~ _ ~ 1nT~ IThe8llia$lb npresentthe chengelI 1n~t1V8 p~lons
ti~~~ [ ~gt 3 - - - _ - _ -
Qf nellllllllt within We surface layars of the pe1ntmiddot ru The
~f~~thtJn -d ~rfeli avr the 336 h11cxI ampScanbe
SII4iIl r~ fig 3 al~ tht rta of~ dirfers tJetwaeo thaa ltmiddot~1- 1ft~- ~ _
IJOl appfiai-ed to ~1nlt1elly until ftar day 168 it fall
~~l~ 1h8relllllrelgn1fi~ diff~be~ qrOtactlid and
proieCted (at -966 ) piIIllnts tor lioo COPPer but not folimiddottin
Ibi~tlons Of the Pain -urface 8hoWad _Pil ~ witilt1M
(Plete I to IV)~ 1hePllat1na (~) pelnt(Phta x and b)
Clearl~st-i~~r alots on the SurfllC8 IiII1kh~ter1sed ~- f hlt1 lriitUl tin conCantrat1on~ lllariy slotsre Partially
~OIIe~ ~ tile _tr~tedal middot1lere rs ~an~ densely packed
grerltr~ ~th 1ar~(iIa1niy cotifIIr) 8ntt -ampniBil (lIB1rily 1rori)being
middot vl~l~~~~~ Dtt~i~~~t~ r~ ~f~~ I~t seawatllr for 11 n IIIOre slots appeared and th~ ~~ess _trb cwer
--if -- V~ c~~ f(i ~~~i~
(Plate Ie to It)~ the granules dld not protrude from the~rtace in ~ ~~~~j1~r~-ffJmiddot0~~
suhap~~r Both 0 nd --960 at ~~pa1nts ~ - fr ~~~~-w1-U~ ~~t
sl~llrat thi stag~ (Plates Ic-d II1d xrnspac~IlYh middotmiddotWith middot middotaJ~cmiddot gt incraallO ~alolre t~ difrrqcbullbull middotbatIiI8IImiddot the protected fjd
bull ~ ~ ~t~
IIllrotacbldpaint ~~lIfIt~ftitr~ h alQt~ p~t bull u bullbullbull J ~~i~~ 1~~
lIIith IIIuper lIdguon u 4lrotatad piIJnt (Pl8ti1 II) 1hD1II8S
Ill )1IMdw1lillS gnnJla~on f-r~~ dpert1cl ~ - - - ~~r ~t
Onthe 966 1IIpetnt t-var tIIO typeaofan with diffannt c t
~hIId~(Plate UI a to c) One hIId tIII-1 bull bullbull e _ bull
fUll wJ1hleas abvlaus alats the o1fwl bed C18UU1IlOt ~litUi middotC c ~ ~
less ~ t11 h Gnnulatlan __ but ~lIiIZly def1nlld in
middot~ latter Afte fila bed tiIIrtIId 10 ~are ~~j(
extansll(tlly ~ the~tIId(O at) tIIIIple lIIIdngthe~ - ~
middot orarallet1ai leas claaI (Plate I~) f u II1ao CQ1tJAJ11d to ~Gn - -
the -966 at 1 (Plata III d tor but 1JI8V8n 1nittNdcneabull -
Althatqa -middotslots wrs abvlaus becaI1ng 1 cwampr aiI- the orwWat1an8
f1nailyftar 14 day exposure ~ fUll drMIlapIIIent on all la - ~
are extensive with the 0 lhawlng both alota pore ~
OVIKgrOloln (Plata II c to f) Gnrulatlon 1bl to ~ gt~t~~ i
or the th1cknats of tho ma ~the ~ at ~ (~~~~ themiddot - - _ middott ~ ~ ~ - -1 1 t
middot f11bull IIIIIS IIOre extans1 thIIIn at dIlya but IIIIIIIY bull1Q~ tI st11l ~w -middotr~ 1gt ~1 ~
obvious fIIany ~ loI8re still p~ yen~~~~~~~r
than before Poiu appearedto be f~lf~ila~~c-~~~a (Plate IVf~ label 1) IIIIl1ch broke~i(2)~ ~~~~
t t
bull ~lS (3) ~ rflC~~ ~i~t~~l~~KI~ t~ldiIiiicyul~loIiih ~~Rht ~ )~- -~ - ~middotd 40 ( gt ~gt ~d lt1 t ~ Jt ~remiddot4amp tha paint trix cavity bottom eroded away reVe81~ng ore
t ~~t~ por~~ ~l~ P~]Y p~~8dby di~lutiaA~ ~~~~ti~Jr~gt~ta(l~~Siinll1cl1Ud~f iUiIiI reriatlGllbullt
bull a~~ ba~tid colCriAtloriccic~ GIl ~~~d ri -966 - ib~~~amp catQn1iti~~t~I~~(~t an ( ~
t the ~~~palnts (PIa_taiiI d~~h ~wh ~ n td have ~ CamplluJMitiIVc) ~~middottti o~lgtnalrf tlt of hIght
~ l=~A ~J~rW tYtles~ p~fiiilriQcOCCold to~Ja to jU-t ~jmiddoti~ai dlatClilS ~ir -jlati ~tbullbullent aNit ~ t ~~ th8middotf~ld~ies~~t~on~ ~ -966 prQ~~~(P14ta Iii~)Omiddotiliof~m1hiiHAiiipieat ere~ll-(piitir~ p-iot~iilans(PiA-iII~ end c) nbullbulltiledt dl~by1i~~ end wirinali1sheii ~cMI~bY thI tili
bull In t-~~ ~1i1nt 1oIIIeHfd~ ~m1IS (15OClind a t ri~r-dlatoa ~lor P~t dlataiOlonlt1anGIl ttw-966
t
bull ~
pr~t~ ~is~ih iion obitJauIJ A ~rot -=1 ~ ~ i __
reprasenttild on the 966 painted 1iUrfc1ncluicunljj ~ rwt
Achnanths Colonisation by bacteria 1n the laboretory bsed
bull eliperlMnt tollMct sbiUar pattern to the fild ~
bull bull Dilicueii
bull bull DUt 1Iidnt ~lona Mve dlttllrint ~~ ictian
It ~ JntM l1tenture ttt Iff yen __ JIll the
bull ~ bicins ani 111 ~t INchlnil (PhillIPs 1t73t EWene~ 1981
In ci1ntlHt tJWY ~tit ~Wl1c iIuCh as 1rbiltyl tirI
~ lite wAlII]ltiClr dlsP8tfu~ middotthiirsttilll~intaJn~ a
bull ~
lIIifoflll ampalCentraUon through the filII 10 thoIt biocide rel_ at tha
paint surfce 1fI$Jlts in IIIOre ditfusing ~s to repla~lj -iflJ~it
resulting 111 bullbulltNdy lr leechlng rata M1llat this IIIIY be middottruIi ot
tin ~ it was nOt true ~ ttw middotlBlf fOZWJlatiGn ua ~ bull - i - bull 1(1
ObSfvt1onundr SE clarly ~ tM t1l1 to be present
~ly lIS OIIIdle-llk parUc1~~t tn lUlfiice (Ito
IV) bUt usa ~ the dIpth attt fU-middot ~y~
If one considra the concn~rtlonmiddot chMge 111 1n ~111c COKICIl8rlts fro- the paint tU- i bullbullbull tMWei b~ ~ Si6t arid
tI-~t iron oxidt 1~ can bt (Fl_ 3 IrId TIibl8I) ~t ~ ase cona1derllble d1fftrenC8s ~ a Un decz bcdYMr ttw p1iod IIIIwraS lron~- an unt Ught inCr h ~ incqJper ~tlGn occUrnd _idly at ttw ~~ Gfb ~ period ~ out ttwn fu nPldly ampQII1n ~ umiddotirid II phast This IIIOUld bt tIlq)8Ctad It cantact l-=h1ng we occun1ng
IIltfIOS8d pertlcl of copper would lMch rapldly on t-n1on 1n - shy JIsewtr There would thlll1 ba bull J-a a1iltUy leu avalUba
i1 ~ ~
_tedal becfM uposed Thi tIOUld ttwn IIllCh Thla pulSing efttct
IIIOUld be expctad to -oth OIItlllith U- putic1 __ -lllible II tot 1~ GIl a on r-x _lao Thla teet 01 put1c1e 1tUng
can tit GIl the lIUrlace ClIlC8 it ~ a pan 1amp t
a 11aM bet in Plat I to IV IIaIrIg _ lit la_ II ~ 0- ~s _- PIate IW $llldl ~bull nshyot poRI flllNltttn Al~ tin ~ lou rete thtn I bullbullbull atill Ivwnce of bull al~t pulsed 1Itue rrlict tII1IIWd
elqlCtaG inIl ttwauUr ~ I I I
0
~ i
~ ~
~I J
bullbullt bullbullbullbullt bull bull ~
bull bull
I
bull t
~~
The same seqiHllle of cevity follliltloii (intttecaseofthetestjjltle
slot- shaped) fClliOileciby SecOlld8rYtlolesfotlllingin thll cavity
bottollis releasing further deep8rtox1ils (Plate XV d and g) As
cavities_re ProdICed ~ tiecaaIelnvadedby_ter~ onelilauld
expect 1ncreaa 1n1IOdha and dIladne lMls t the t1met
which the toxlM uw ~ Ths fCMld tCtHi eo (Flvmiddot 3)
~rttno the vi that_tat ~t1on otthe~ ~lY
il1Cl1l8S8S Idth UIiJ
Differences f~in toxin li~~to Ini 966 ill 8811P~ F ~ bOth Un end ~ lcMIr levels afillW
IIIIIlfI ptaInt(~1~~1ntha~~ ~~f~~ cst rklld with tbiIccpper ElJIc~l ta~Uona invalvinv
w pl8cId at a~ht~~middot3lO~lth Un cqJpel bull
(-960)10 ttW ~af this expei1tIIri1ts thIsiorelfJltdt -
that cqlp8r b8IiIId ~ IIiIOUldProc-ln f41ip1dly t thellllgllitiv
patentW _lied () as 1t 11 further f~1t ~ witt Un
III11ch is u-tat aqulllbd Idth
it If ttda 18 eo~ ~
~ )
frOll1 paintscould bl anqnced by an illprd current clithodic
protecUansysteipos1bly ahart_nino te 11fe of the paInts
antifouling ability Bacterial coloni Uon IIIU rtdUCed on the -s66 paint This
could have been due to unflSllOUlllble electrostatic cqrge on th surtece Th1a effect CIIl pley en IIIportant role in the t~of
bacteria to alrraces (lIIarsMll19E1O) As bacteria appear to IhiJw
greater toxicltyto copper (DMpsey 1981) it is Klre likely that
enhanced cqJper leacll1nl On the -966 AIIIPl c8used the recluCed
activity The IDlol8r toxicity ofUn to bacteria (as reported by
~ -
DelIIPsey Hle1gt or its lowes leaching rat was sUpPorted bythl
relatively rapid recolonisation of the slots by lIIicroorganiSlllS ~ftar thE inlt1al aurge of leachi~ Thailllpressecl current dto ~~ the oppoSite _ff~middot ~r on dtta colonstJon l1lI-1- in the
JIbcJrtory~ta larger rUDIIra f~ fit pro~td ~ther than uriprotected Pafnta
Ccnlus1one
1 tiild-a 18 presaIIUd ~ thattJn~ ~~
~llNChng notd1rf~ ~
2 TIleuseofCP~ ~CUIftnt(at) on1h1a ~fca41ng ~- ~
pa1I]t~ to ~_lAIIach1ngn_ at toxiN - -
3 leIJstaPidbectar1al ca1on1aatJon acaampmId an ~bId Pt1ntJ ~
4 JItore larger alcrofauJing (Oiat_ and PzOtoaoana) Gr~_
occurred on the protcbld~
I l
I
bull bull ~
mfEflpoundaS middot~-t9i _jgt- ~ _ _ ~sev M~J~ (1981) (cilonJzatiooof alUf~lng lnts by _rine
Jbull ~teda9g~1ta IIarina XXlV~ 185-191
- - gt 0 Ie 1 _ - _
poundV8I$i I~lf (1~P~h 1II8rin~Ugae and Foul lng AReviw ith
t p~l-i9Jrlr ~iiMdilto5hiP-fQUllfCiH 8ot~ ~r~ ~~~ XXIVbull lIPwl~J11A
Fotteth CN~R ~kk8nGB IWPIR1seJCiiiroaic1lihd ~e ~JIIth CII Offshore St1ucturu ElllS ~tjI~f~1Dt19i4
- - ~
UKC (1976) I~races ill ~W Ecology KIuverd Univbullbull middotraabullf~iddge ~bull C~~jII illt)Ia(t RC (1980) totinllsa Cattod1CPrOtect1cinbull middotmiddot8AtEmiddot~6haOCIlfif~ T~ p No 108 ~ 18 ~ 1IIHf bull
t bull lt ~J~~~ __ ~
Norkr4irulB(198D) Advanculn UcrOfj~lpound~ltiOyVol 4 (AlItxlaquoIder fl Ed) PlerIw Press lcxIdon~ lIP~ 51-85 ~ l~l - bull bull
PJijllt~i At(fm)~mr~~JnI~1fqul1nQ JlalntlJ Riumh ~JnOrpnic toatJngs 2 192middot
)~J gt0 c ~lt -- ~ __ RogerS~ Ut (1968) ina ~(lnt~fbItliP4ili GI COnoslon ~a Technology)bull bull
Sl~iorr~ VP~Ranald~ Rctd~~ (1S80)_ ~ Experllattal Stud1aiRelate middotEffect or Cathod1c Protict1Oi withmiddot terta1nr1c
Coating Syst~ 12th Annualofr T8Id1na1ogy Co1fItHnC8 liauston TIIlt p 9-12
j
fIG1 sPct FromMon IT8TJCuZOI Int Lliu EOAX
FlG~ Fro TlTCu20Ar-4 Dy Field Ellposult I~ir
il
I c
R hi -00amp=7
-I I I I I I I I I f
iii
bull bull
bullbullbull
I J
J
I Figur 3 Thl chan In reltl rqpoIQnlof -ariOtimiddotjOmponents of the paint film with time Thl vtic~1 ampkiS unl till _n ofmiddotmt Illment per unit OJ fe 50all Cl Na
Cu StI Feeshy
PlistinllFe S7~J6 704 3~74 908
f
r e ~ ee~
middotie ~lt - ~ shybullt bull ~
t -~ l Cu~ o ~ f
t --- 3igt - bull 0 J Sn
cc
t ~
~
bullo
~ bull e
bullbullbullbull bull ~
bull ebull a QI 1- bull 0 40 431282 316
101 dmllIII)bullt bullbullbullt
hr 0 ~~bulla 5~16 ~~ -~~ i166 4S i83 8~1B
24tu 6~480 SD~83
111middot 1~1 OS6
~ 5114 6~69 -51 UIamp middot016- lt ~rmiddot
1d o Mea 818 21 021
~Z21 199 1~1Qmiddot ~Jgt2 058
14 d o ~~ 804 Q~B8~~~a~~t~middot 685
866 372amp 8amp4 OS _i~11[ ~~64
Tttile 1 A81UvlL pzQJJortlcw 0 vxl~oI ~~bull Jn 1I1Dtitctel lt-1166) end lqIlDteCt6 (O)middot~af~_~middot_lodEDf~ middotmiddotmiddotmiddotmiddotmiddotmiddotmiddotmiddotc
~ ~
_----I~
J
I bull
I
J bull tmiddot
7o CONQRESOIMTERNACIONAL
I D~CORRO$IOr(IAAINAEmiddot~USTACIONES~v c)
t 7thflfERHATlONAL CONGRESS
ON IARINE CORROSION AND fO(JUNG
~CONGRE$INtERNATIONAL DE~CORR~ON ~E ET QES~SAUSSURE$
i t
~ ~
bullt
t t
UNIVERSIDAD PourtcNlCAVollncio7J1 NiwiemIif~ 1988
(SPAN
SECcl6lfn
BiOIogro marino Morine biologY ~1i1orine
Theunpub1ishedJllBIlu8criptEl have been reprod~d asr~ielved from the
authors The SCientiUc COlIIIBittee take not respoi~ulbll~~Y for any shy
error or omssi9n
Laa ponenciasno han side publicadas y estanreproducidas tal y como
sellan recibido demiddot BUSmiddot autorea pound1 Conte Cientfico middotno se responsemiddot
bil1za de cualquier error ~ om1si~n
bullbull _ bullbull_ bullbull ___ bull $_ _ L _ - _ --- -__
SESSION II Marine biology
Friday 11th November
SURFACE PROPERTIES ON NONmiddotTOXIC ANTIFOULING PAINT FILM
K Nanishi M Murase Y Yonehara M Kishihara and T Hirama
MICROFOULING ON ALUMINIUM PANELS PLACED IN THE TROPICAL WATERS
N B Bhosle K Venkat and A B Wagh
EFFECT OF TRIBUTYLTIN OXIDE (TBTO) ON THE OVARIAN MATURATION OF THE PRAWN CARIDINA WEBERI
R Nagabhushanam B Indira R Sarojini and Mary Frances Thompson
EFFECT OF BIOACTIVE MATERIALS FROM SOFT CORALS ON BACTERIA ASSOCIATED WITH COMMON FOULER
Vitalina Mary Avelin Mary R Sarojini and R Nagabhushanam
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
bull bull
~
bull - -- ~It Jf bull
- ~ 13
12
Terry LA I~Ed)VeangtRfiJ 1981 lIi(toal~IOI1CGrmianF~ l)i 1St1oJhemailnEifooiiiigffixed gftshoreIt Bot 1Iar ~ 117-183 lsect~~jlaUoQ1l1 _ ~t Ef)e~gy Offs~r1I Te~lOtjjy Paj)8r
-middotltcntpI)~it$t~l91-16PP~middotmiddotdl ( I
bullbull
iIIalk e1 JL 1971 bull Ion specfie Uquid I~~ e)(charig~ ndcroelactroclels Anal ChIIaI ~8S-Q2 AGraf1~ w~S 9S1~ 1orrosloh InFIntroooctI00 tooffshOle
51~weSoes~~fab~lcat~onIns~allat1on Gulf Wall PI aelUnger EG 1982 The appl1~tlon 9fMl1ikIngmiddotHoust(jfW~Q23g-258middot icroelectrade in the study of pH end PD2 in lIHinli
HerbierC SN fGH~rt~ )RI -Stoctcdalpii Watltif1Sal RS ~1ronIIInb Ub~~ 27l-Z79 1985 Fac~or lontro1l11)9 tha activity ofmiddot sulpMte
bull Wall P 1983 The appllcatlon Md ~ ofiwrJ)e~c~odl shy-taicfii9 tiiCtetili tn~ diiilncj tar infection
techniques for the iiaurnt cif icraeov1ror1liltital5QiCiety ofJebo1- Engi~ SPE 139781-10 _i~~ - ~ ~ i ( _ IKIdif1catlons caued by aGII8 rlnefou~lng -C~ebullbull
PhD lhaaia Ibiverslty or ~tar UKbull bullbull bull -
IIH1nke JA 19fiSa In EXperllRants in Physiology and middotampioClIiIiiilatJift l c E(If(orltGAiKetkutgt ~c-Presa-
Wooltdngton AObullbull Davenport J 1983 pH lind IlIl2 IMla ffl~- j~t~5 -lMIIleath 1IH1na ro-fauUngGqWl J bull Expbull 1
B101 Earlsectsect 113-124 Klnke JAIII 1969b II) Glass IUcroelectJQdes poundx1tors Uwal1~Ui~ OrOllt arid~t Nt JOhn fII11ey YOI1lt thaptar 11
~-f _f~ ~ ~~nt ~ Igt
bull Iverson IIIP 1972 Biolog~ Qrroaian In Advencesin
corrosion scianca and tecIYIology 2EdS FCntaia G~ I bull IIS~et RIIIbull p~ PJaSsLancIoIl 1-42
bull
Jorgensen BB Reusbech NP I Cohen T 19B3 Photosynthesis and strUcture of bIIn~ II1crobWp MC1OIIlectrocki and ~ stud1ea of four cyanabacledalt -~usllWtOl ~bullbull 1~f093
t l~ O~ ~~~ H~ ~9s7~c-gtHatste~iecfn1k van shy pellediftlrtenPt - Stlchelektraden (1-Su
AiJBitiKlificfiliilislflilr)zlIr polrapusdiailJIIissui1g_ liIserstoff(jruQkas fur d1eB lIsUbullbullung der IIliJlttOUf1cillatlon PflftgaHArdI ~fl39
bullbull Mller JOA 1981 tals In EconcinI1c II1croblolooy sect McNbUdbicldatariOtationEd Rosebull A middotAcadelc Praulondcn149-202
t Mller J DAI King RA 1975 8io$~iorat1on of lllltals In lIIi~1al ClflPlCts of the deterloration of tarWbullbull EdsGilbertRJ~ middotloveloCk 0111 bullAcadeic- Presst IILandon 8Jo103
bullt
IPotgate JR 1976 The sulphate-reducingbacteda
Callbridge tIn1versity Press Cliilbrldg8U-K
Reusbecb fUI i Ward DIII 1964 i~~~1Ida5tud1el of
bullt inter$Utilll IIIItez dttm1stry and photosynthetk activity
in ilflaI spring Idcrob1altAppl~EIi~ MerobW tsect4IlU-27lt I
IIbullt bullt
bullbull Ibullt
1
Inmiddot middotmiddot11middotmiddot f~~ a~ -tmiddot bull J~f
I~ W
i5iImiddot J~a t
ilmiddotmiddotmiddot I middotIt Ii
l
~
middotmiddot~~middotlL= g r
Lj Ic
t
i ~
I
gi II cEi ~ I I
1 t rW
D
tR bull
bull t 10
11
r
i
I ~
j
~
I
~bullbullbullbullbullbullbullbullbullbullbull
LV
FigS pH and p02 profiles into macrofoltling eomlllllllitilS dominated by Balm trenatU$ on steel plate 11 two pH profiles Ib) two lIOai profiles leI pH and P02 profiles into dead barllllCle shiella withdetri~Us I I
itIbl r p02 ~ I
nvnHg 11 70
j ifJi eo 1amp0
laquof it 30 s
I 20 N
ca 11 10 a o -- shy i bull
11
80
19
781 o 2 6 8 10 12 0 2 6 amiddot 10 12 dIpdI Qf ~iftto fouling In HighIst numbIII pIatIi
8deg~~
~i c5 iioo5 fiil
pH 8 bull_ ~
~~ ~
_ bullbullbull- pH
-pOz
(cJ
e-Clf ( j~JJlaquo1- --~I ~a ilil - _ I i ]1
~ ri en i ~ oJt ~iJ id tUI
2u1U I shyI t
shy1 2 3 6 6 7 8 9 10 11 12 i shyDepth 9f electrode (mm) into fouling
1 i
1
~----------~-------
~
~-~ lt1~
middotmiddotPmiddot ~
I
V
~
ill
1 Tbellitt~~ of cathodiC protection OO~~lIic~tlO aridrfOflllUlc
ofi AA~Uoullr14l ~1~I
I
ael~lner EG middotIUtcb8l1AJ Dept lt4 lInY1roMe1lal Bl01011
=~~venilt7 IlariCheW 1f139PL $ ~11al1df
r
I
Ablitrict ~~~ e~l billttI ~ted with ~pert1n ant11_1111ampp paint llere itpoii~1n _vatI- In the tl~ld iUIit 1bOioatbrt~ScM Were
1IUbjll~ to qtbocJ~C ~teltt1on byen _ current (~)SJi inci EDAX Iieamp~ta IJdlctecI ~ lachln teoc copper OIl tbePlCtec~ pa1n~ atter14 dQ1a~ protected Pllnt abo ebowed
) reduCed bacteiial activIty butmiddot ~e4 dltoe Iu1d protOzoanmiddot co~~l~tlont~1l IlU l~byenCOCltact not byen dUfwIl00 ~r _baRl lrillolvedln tOXln nil ereio noted
l ~~
1
I
Introduction
Applications of protective coatings and cathodic protect1on(cp~l
are recognised corrosion control methods for IIIBrine steel Itructureso Cathod~c protectlOll preventl conCIsion by Int~lng til8ctrl~l
currentS trem externalllClUfCa to e~~tthlJno~ e~ carrollon feetlans C04U lUChu tUauJ1ng Aelnts a
(
barrIer to ~ the flao of couos1a1 ~aboIHnu~r
Dccurrlng anodes cathodes or wlwit gelvWc CCIIII1 ( 19681
flinger 1980) AnUfauling ~ also JJICOqIOrII~ b1oc~~
Tr~ylt1n nuor~ (~) end o~ Oxide (~) (~ flJ1J
- of u blacld1111~
V _Ut Evans 1981) ~ ~
end growth of bipfoullng 0I1I)IIIII1IIIIe Wdd~ ca ~ of
paint rusendmiddotmanemiddot ~(r~ til )
As corrosian ~t1on ~ bICCIIIII -an ~1tlQd ~ of
antiroullnQs 1 conjunction _ith CP have bllClIOr ~
resultIng in reseudt 1rwestllj111tlng trw COIIIPIitlbUlty or VU~ ~t t _~
rOratlation to dlfrert levI ot CP (~Eelf 18110 ~
~t 1980) RelatIvely 1nIestiljlllUans have ~ urtw ~
or CP an thIt anUraullngpert~ of palnta
In thIt preseot 1nlIestioaUon tV~iln111 ronutlon ud
cOIIIHrclallY incanjlntlon wIth CP was ~t4IC~ Iw I sraed current (-966 l1li) arid exposed at the 1 Strait IInQlbullbullbully
~t1ons weremiddot 00 the IIhcIrt bnifects the PprIUd_
had an a) the various COIIIPQI1eIIts of n pe1ntf1l11 end b) l1oft~
developaentbull
bull bull ~
~ ~ - f
bull ~ _T~~-middot~ ~~- ~r _ -$1
(1) ~rlttionor Painted Steel Billets middot_ ~~tmiddot_tlJ middot~hmiddot~--~fmiddot ~ middot~A~-~~middot - -~ ~
Thirty-al ateel blllets (8-10aD2 end l depth) were cut frollbullt ~middot~middot~middotmiddot~middottrSmiddott ~jS-~ It bull gt~ tfl - middot--imiddotmiddot ~ t1dItrd steal plates The dimenSlonsof -thB butits-lim chosento
bull -~~ middotmiddotgt2h ~ ~-Imiddot ~~rt ~- -middoti~~ ni ~~middot1 l middot-~t~~middot _
(a) fecilltate amiddot Ie detrucUve -thxI of prePraUonpdor to ~~_gt gt~ k7i ~_ t~ 1--1- _ ) - +--_~ -_~bull ming electron ~ 4II1CI (iialloWdirectobs8iVatlCnS of the
- i lt-- ~ gtbull -o1e billet 8Uface
gt r~middotmiddotj ~L(~middot~middot rmiddot tmiddot J ~ - ~ - lt _ bull Single non-standard (1hIula~) nickel lIIireSmiddot Spot Welded e1Cing
bull ~1deJ ~ b~i~ ~j~~ tri afl~1b1ipOi~ f ~1t1 ~4middotmiddot f-gt -~ ~S-- ( -- - - _bull resin EactI billet then Undb~ tI1d prtiladlllltha5O __ eyilr
bull ~~_ ~~imiddot ~1iil h~ intlfcili~ ~eonbt1nlyi Tr1butylt1n F~id(18n)~~~O~dettu20i~blcicld8is 1IIfii bull i - ~ ( then 1IPI)l1ed to glvaadry Ulil tIi1c1ltnOss or 1(1)-150bullbull
tbull (2) IipreSSeCi Current Syat
bull A rigid ~tic due 1I8S drWedllliihwrtv-s41 liiIihOieS IIPProX~
bullbull 05-10 cia episrt ~ Pilnted ampUl8t Wsfbed IntaPQSIUon ampy
threadlng the nickel IIIlr8throu11 the dinto a tarproot junction
box Eliltilen of th8~1l1ii (recelving ~t-d curnritlere IIIlred
Into bull block connectOrrecelvlng dc trOll is potentiOstat The
rainJng iontroi blUets Wet8fbedfl11i1iiy lIIithln thejtlytlonbox
i~ityd~c IkiliPly ~~ ~lat1ruaWiode 8ncIacalOliel
refer~ electtodetoiere fllied ~ the per1aetat of the plut1cdlc IIIId
connected to the pot8ritlostat The desl~ of the potentlosbt ailoiolEld
~PlDteItlO1TClJrnnt to be MIllntained at 8prampset levtl of 966 II1II
c
r~
Grid raf 1J5s-t 114iH 563724 dud 0 Juna
Alolecule andbechr1a in general are ~ to
sutface lI1eroleyer (nlUSton) of _ter (~bullbull 191KJ)~To avold
contaJnatlonof ttw painted surf by U- -vents
blllets Were s-nact in a 1 conte1ner or atc1le (02i ) f~~nd
ter Iih1ch then f9ICI in tv 1IIhere the b~
be r-oved undeNeter El~~ bllleta fa Mcb trtai bullbullel
removed over a 1 dtq (336 hr~ expoeure perioda
each being ~~bn uposurnof 1 5 11 iA 168and DlIIauz1r
Arter field axpauHli the billets wan
fUtered _tar to ~ 1_ debtlflMd far 10 aJnuta ~
wv ~~~endattid sdbUiUd(otO
~ ~l f~ nn1ng el~~ --
lOOted onto 1middotCIi ~bullbullWIsend carbonmiddotcoa~nmiddotmiddot~middot f
each billet Ws~ Iis1ng a ccr1dge ~el~~
(5360) IIIlth anergy dJ8perslve X-ray alcroenelre1a (mx)
_lysis the les Wen reNed end coated III1th vold to al]Ql En
detailed sat of the alaquofa1l
(4) Lfiborattux ~
Painted taa~ bUletalllllre pnparad ~
in a Mrlne reclrculet1ng flOlll _ ~
f1Oll the ~ ~a1t~y Tha~~Un bullybullbullbull
10C M1nta1ned ~~t1as of ~t~la protazOllbull~bull10shy
(Entermap) ~ ~~~middotNlt~Eamp1~ __~and ~ r - ~ -
certaJn ~lc 1IQ1~ 1(1 the
the painted
~
thre repl1catft fn
plKed 1nto(022_)
- thIine1J~~ -lt bull
Atter mAX
(1-2) and i-n-t bn collected
14OC
Pain~
-
12 daY~ exposure and prepared as in d)~ - t
~I ~~~-- i~yen 1~ v
Rflltsi ~ _ ~f-~lI ft j bull _
rt~ll0~~~llY~~~~peln~stetsl~11i~ex~sed for
y~t1~J~ ~~~~1~1~~1UII~ya~kooJ~~~lt ~ ~~_~~I1J~ ~~~tomAX -lysis ~era than gold ~ bull bullbullbull c - _ bull lt
~~fllr ~~~ vllUll ~t1ans of the rfee htures and gt Co
~f~ - ~~~ ~f ~ ~aobta1Mdue IjIll811 iI1Flg 1~2 The IIIIdn ~~ ~ lt)ji9 )tmiddotmiddot - ht
- bull
~I rIIplt~ tIt8 -~ cOlllponanta of the pei1t 1bullbull WelIIlItr1x $ilj~t-~~middot - ~ _ -_
-IUch PVCbQed end rap~13y the chlorlna peak the
i~~~-~~-~V~~~-t18 ~~~~ ~~ Th8
ral~~i~ JlrQP(lrUo1sof ch 1Il-ntal paek~ll1d l~t_ Q~7 -gt- ~ _~ -- gt ) gt -_ -lt ~~ the 0 -end _ xprtsntabullTIwsa changes are -r1sed ~t~ lt~ _ ~ 1nT~ IThe8llia$lb npresentthe chengelI 1n~t1V8 p~lons
ti~~~ [ ~gt 3 - - - _ - _ -
Qf nellllllllt within We surface layars of the pe1ntmiddot ru The
~f~~thtJn -d ~rfeli avr the 336 h11cxI ampScanbe
SII4iIl r~ fig 3 al~ tht rta of~ dirfers tJetwaeo thaa ltmiddot~1- 1ft~- ~ _
IJOl appfiai-ed to ~1nlt1elly until ftar day 168 it fall
~~l~ 1h8relllllrelgn1fi~ diff~be~ qrOtactlid and
proieCted (at -966 ) piIIllnts tor lioo COPPer but not folimiddottin
Ibi~tlons Of the Pain -urface 8hoWad _Pil ~ witilt1M
(Plete I to IV)~ 1hePllat1na (~) pelnt(Phta x and b)
Clearl~st-i~~r alots on the SurfllC8 IiII1kh~ter1sed ~- f hlt1 lriitUl tin conCantrat1on~ lllariy slotsre Partially
~OIIe~ ~ tile _tr~tedal middot1lere rs ~an~ densely packed
grerltr~ ~th 1ar~(iIa1niy cotifIIr) 8ntt -ampniBil (lIB1rily 1rori)being
middot vl~l~~~~~ Dtt~i~~~t~ r~ ~f~~ I~t seawatllr for 11 n IIIOre slots appeared and th~ ~~ess _trb cwer
--if -- V~ c~~ f(i ~~~i~
(Plate Ie to It)~ the granules dld not protrude from the~rtace in ~ ~~~~j1~r~-ffJmiddot0~~
suhap~~r Both 0 nd --960 at ~~pa1nts ~ - fr ~~~~-w1-U~ ~~t
sl~llrat thi stag~ (Plates Ic-d II1d xrnspac~IlYh middotmiddotWith middot middotaJ~cmiddot gt incraallO ~alolre t~ difrrqcbullbull middotbatIiI8IImiddot the protected fjd
bull ~ ~ ~t~
IIllrotacbldpaint ~~lIfIt~ftitr~ h alQt~ p~t bull u bullbullbull J ~~i~~ 1~~
lIIith IIIuper lIdguon u 4lrotatad piIJnt (Pl8ti1 II) 1hD1II8S
Ill )1IMdw1lillS gnnJla~on f-r~~ dpert1cl ~ - - - ~~r ~t
Onthe 966 1IIpetnt t-var tIIO typeaofan with diffannt c t
~hIId~(Plate UI a to c) One hIId tIII-1 bull bullbull e _ bull
fUll wJ1hleas abvlaus alats the o1fwl bed C18UU1IlOt ~litUi middotC c ~ ~
less ~ t11 h Gnnulatlan __ but ~lIiIZly def1nlld in
middot~ latter Afte fila bed tiIIrtIId 10 ~are ~~j(
extansll(tlly ~ the~tIId(O at) tIIIIple lIIIdngthe~ - ~
middot orarallet1ai leas claaI (Plate I~) f u II1ao CQ1tJAJ11d to ~Gn - -
the -966 at 1 (Plata III d tor but 1JI8V8n 1nittNdcneabull -
Althatqa -middotslots wrs abvlaus becaI1ng 1 cwampr aiI- the orwWat1an8
f1nailyftar 14 day exposure ~ fUll drMIlapIIIent on all la - ~
are extensive with the 0 lhawlng both alota pore ~
OVIKgrOloln (Plata II c to f) Gnrulatlon 1bl to ~ gt~t~~ i
or the th1cknats of tho ma ~the ~ at ~ (~~~~ themiddot - - _ middott ~ ~ ~ - -1 1 t
middot f11bull IIIIIS IIOre extans1 thIIIn at dIlya but IIIIIIIY bull1Q~ tI st11l ~w -middotr~ 1gt ~1 ~
obvious fIIany ~ loI8re still p~ yen~~~~~~~r
than before Poiu appearedto be f~lf~ila~~c-~~~a (Plate IVf~ label 1) IIIIl1ch broke~i(2)~ ~~~~
t t
bull ~lS (3) ~ rflC~~ ~i~t~~l~~KI~ t~ldiIiiicyul~loIiih ~~Rht ~ )~- -~ - ~middotd 40 ( gt ~gt ~d lt1 t ~ Jt ~remiddot4amp tha paint trix cavity bottom eroded away reVe81~ng ore
t ~~t~ por~~ ~l~ P~]Y p~~8dby di~lutiaA~ ~~~~ti~Jr~gt~ta(l~~Siinll1cl1Ud~f iUiIiI reriatlGllbullt
bull a~~ ba~tid colCriAtloriccic~ GIl ~~~d ri -966 - ib~~~amp catQn1iti~~t~I~~(~t an ( ~
t the ~~~palnts (PIa_taiiI d~~h ~wh ~ n td have ~ CamplluJMitiIVc) ~~middottti o~lgtnalrf tlt of hIght
~ l=~A ~J~rW tYtles~ p~fiiilriQcOCCold to~Ja to jU-t ~jmiddoti~ai dlatClilS ~ir -jlati ~tbullbullent aNit ~ t ~~ th8middotf~ld~ies~~t~on~ ~ -966 prQ~~~(P14ta Iii~)Omiddotiliof~m1hiiHAiiipieat ere~ll-(piitir~ p-iot~iilans(PiA-iII~ end c) nbullbulltiledt dl~by1i~~ end wirinali1sheii ~cMI~bY thI tili
bull In t-~~ ~1i1nt 1oIIIeHfd~ ~m1IS (15OClind a t ri~r-dlatoa ~lor P~t dlataiOlonlt1anGIl ttw-966
t
bull ~
pr~t~ ~is~ih iion obitJauIJ A ~rot -=1 ~ ~ i __
reprasenttild on the 966 painted 1iUrfc1ncluicunljj ~ rwt
Achnanths Colonisation by bacteria 1n the laboretory bsed
bull eliperlMnt tollMct sbiUar pattern to the fild ~
bull bull Dilicueii
bull bull DUt 1Iidnt ~lona Mve dlttllrint ~~ ictian
It ~ JntM l1tenture ttt Iff yen __ JIll the
bull ~ bicins ani 111 ~t INchlnil (PhillIPs 1t73t EWene~ 1981
In ci1ntlHt tJWY ~tit ~Wl1c iIuCh as 1rbiltyl tirI
~ lite wAlII]ltiClr dlsP8tfu~ middotthiirsttilll~intaJn~ a
bull ~
lIIifoflll ampalCentraUon through the filII 10 thoIt biocide rel_ at tha
paint surfce 1fI$Jlts in IIIOre ditfusing ~s to repla~lj -iflJ~it
resulting 111 bullbulltNdy lr leechlng rata M1llat this IIIIY be middottruIi ot
tin ~ it was nOt true ~ ttw middotlBlf fOZWJlatiGn ua ~ bull - i - bull 1(1
ObSfvt1onundr SE clarly ~ tM t1l1 to be present
~ly lIS OIIIdle-llk parUc1~~t tn lUlfiice (Ito
IV) bUt usa ~ the dIpth attt fU-middot ~y~
If one considra the concn~rtlonmiddot chMge 111 1n ~111c COKICIl8rlts fro- the paint tU- i bullbullbull tMWei b~ ~ Si6t arid
tI-~t iron oxidt 1~ can bt (Fl_ 3 IrId TIibl8I) ~t ~ ase cona1derllble d1fftrenC8s ~ a Un decz bcdYMr ttw p1iod IIIIwraS lron~- an unt Ught inCr h ~ incqJper ~tlGn occUrnd _idly at ttw ~~ Gfb ~ period ~ out ttwn fu nPldly ampQII1n ~ umiddotirid II phast This IIIOUld bt tIlq)8Ctad It cantact l-=h1ng we occun1ng
IIltfIOS8d pertlcl of copper would lMch rapldly on t-n1on 1n - shy JIsewtr There would thlll1 ba bull J-a a1iltUy leu avalUba
i1 ~ ~
_tedal becfM uposed Thi tIOUld ttwn IIllCh Thla pulSing efttct
IIIOUld be expctad to -oth OIItlllith U- putic1 __ -lllible II tot 1~ GIl a on r-x _lao Thla teet 01 put1c1e 1tUng
can tit GIl the lIUrlace ClIlC8 it ~ a pan 1amp t
a 11aM bet in Plat I to IV IIaIrIg _ lit la_ II ~ 0- ~s _- PIate IW $llldl ~bull nshyot poRI flllNltttn Al~ tin ~ lou rete thtn I bullbullbull atill Ivwnce of bull al~t pulsed 1Itue rrlict tII1IIWd
elqlCtaG inIl ttwauUr ~ I I I
0
~ i
~ ~
~I J
bullbullt bullbullbullbullt bull bull ~
bull bull
I
bull t
~~
The same seqiHllle of cevity follliltloii (intttecaseofthetestjjltle
slot- shaped) fClliOileciby SecOlld8rYtlolesfotlllingin thll cavity
bottollis releasing further deep8rtox1ils (Plate XV d and g) As
cavities_re ProdICed ~ tiecaaIelnvadedby_ter~ onelilauld
expect 1ncreaa 1n1IOdha and dIladne lMls t the t1met
which the toxlM uw ~ Ths fCMld tCtHi eo (Flvmiddot 3)
~rttno the vi that_tat ~t1on otthe~ ~lY
il1Cl1l8S8S Idth UIiJ
Differences f~in toxin li~~to Ini 966 ill 8811P~ F ~ bOth Un end ~ lcMIr levels afillW
IIIIIlfI ptaInt(~1~~1ntha~~ ~~f~~ cst rklld with tbiIccpper ElJIc~l ta~Uona invalvinv
w pl8cId at a~ht~~middot3lO~lth Un cqJpel bull
(-960)10 ttW ~af this expei1tIIri1ts thIsiorelfJltdt -
that cqlp8r b8IiIId ~ IIiIOUldProc-ln f41ip1dly t thellllgllitiv
patentW _lied () as 1t 11 further f~1t ~ witt Un
III11ch is u-tat aqulllbd Idth
it If ttda 18 eo~ ~
~ )
frOll1 paintscould bl anqnced by an illprd current clithodic
protecUansysteipos1bly ahart_nino te 11fe of the paInts
antifouling ability Bacterial coloni Uon IIIU rtdUCed on the -s66 paint This
could have been due to unflSllOUlllble electrostatic cqrge on th surtece Th1a effect CIIl pley en IIIportant role in the t~of
bacteria to alrraces (lIIarsMll19E1O) As bacteria appear to IhiJw
greater toxicltyto copper (DMpsey 1981) it is Klre likely that
enhanced cqJper leacll1nl On the -966 AIIIPl c8used the recluCed
activity The IDlol8r toxicity ofUn to bacteria (as reported by
~ -
DelIIPsey Hle1gt or its lowes leaching rat was sUpPorted bythl
relatively rapid recolonisation of the slots by lIIicroorganiSlllS ~ftar thE inlt1al aurge of leachi~ Thailllpressecl current dto ~~ the oppoSite _ff~middot ~r on dtta colonstJon l1lI-1- in the
JIbcJrtory~ta larger rUDIIra f~ fit pro~td ~ther than uriprotected Pafnta
Ccnlus1one
1 tiild-a 18 presaIIUd ~ thattJn~ ~~
~llNChng notd1rf~ ~
2 TIleuseofCP~ ~CUIftnt(at) on1h1a ~fca41ng ~- ~
pa1I]t~ to ~_lAIIach1ngn_ at toxiN - -
3 leIJstaPidbectar1al ca1on1aatJon acaampmId an ~bId Pt1ntJ ~
4 JItore larger alcrofauJing (Oiat_ and PzOtoaoana) Gr~_
occurred on the protcbld~
I l
I
bull bull ~
mfEflpoundaS middot~-t9i _jgt- ~ _ _ ~sev M~J~ (1981) (cilonJzatiooof alUf~lng lnts by _rine
Jbull ~teda9g~1ta IIarina XXlV~ 185-191
- - gt 0 Ie 1 _ - _
poundV8I$i I~lf (1~P~h 1II8rin~Ugae and Foul lng AReviw ith
t p~l-i9Jrlr ~iiMdilto5hiP-fQUllfCiH 8ot~ ~r~ ~~~ XXIVbull lIPwl~J11A
Fotteth CN~R ~kk8nGB IWPIR1seJCiiiroaic1lihd ~e ~JIIth CII Offshore St1ucturu ElllS ~tjI~f~1Dt19i4
- - ~
UKC (1976) I~races ill ~W Ecology KIuverd Univbullbull middotraabullf~iddge ~bull C~~jII illt)Ia(t RC (1980) totinllsa Cattod1CPrOtect1cinbull middotmiddot8AtEmiddot~6haOCIlfif~ T~ p No 108 ~ 18 ~ 1IIHf bull
t bull lt ~J~~~ __ ~
Norkr4irulB(198D) Advanculn UcrOfj~lpound~ltiOyVol 4 (AlItxlaquoIder fl Ed) PlerIw Press lcxIdon~ lIP~ 51-85 ~ l~l - bull bull
PJijllt~i At(fm)~mr~~JnI~1fqul1nQ JlalntlJ Riumh ~JnOrpnic toatJngs 2 192middot
)~J gt0 c ~lt -- ~ __ RogerS~ Ut (1968) ina ~(lnt~fbItliP4ili GI COnoslon ~a Technology)bull bull
Sl~iorr~ VP~Ranald~ Rctd~~ (1S80)_ ~ Experllattal Stud1aiRelate middotEffect or Cathod1c Protict1Oi withmiddot terta1nr1c
Coating Syst~ 12th Annualofr T8Id1na1ogy Co1fItHnC8 liauston TIIlt p 9-12
j
fIG1 sPct FromMon IT8TJCuZOI Int Lliu EOAX
FlG~ Fro TlTCu20Ar-4 Dy Field Ellposult I~ir
il
I c
R hi -00amp=7
-I I I I I I I I I f
iii
bull bull
bullbullbull
I J
J
I Figur 3 Thl chan In reltl rqpoIQnlof -ariOtimiddotjOmponents of the paint film with time Thl vtic~1 ampkiS unl till _n ofmiddotmt Illment per unit OJ fe 50all Cl Na
Cu StI Feeshy
PlistinllFe S7~J6 704 3~74 908
f
r e ~ ee~
middotie ~lt - ~ shybullt bull ~
t -~ l Cu~ o ~ f
t --- 3igt - bull 0 J Sn
cc
t ~
~
bullo
~ bull e
bullbullbullbull bull ~
bull ebull a QI 1- bull 0 40 431282 316
101 dmllIII)bullt bullbullbullt
hr 0 ~~bulla 5~16 ~~ -~~ i166 4S i83 8~1B
24tu 6~480 SD~83
111middot 1~1 OS6
~ 5114 6~69 -51 UIamp middot016- lt ~rmiddot
1d o Mea 818 21 021
~Z21 199 1~1Qmiddot ~Jgt2 058
14 d o ~~ 804 Q~B8~~~a~~t~middot 685
866 372amp 8amp4 OS _i~11[ ~~64
Tttile 1 A81UvlL pzQJJortlcw 0 vxl~oI ~~bull Jn 1I1Dtitctel lt-1166) end lqIlDteCt6 (O)middot~af~_~middot_lodEDf~ middotmiddotmiddotmiddotmiddotmiddotmiddotmiddotmiddotc
~ ~
_----I~
J
I bull
I
J bull tmiddot
7o CONQRESOIMTERNACIONAL
I D~CORRO$IOr(IAAINAEmiddot~USTACIONES~v c)
t 7thflfERHATlONAL CONGRESS
ON IARINE CORROSION AND fO(JUNG
~CONGRE$INtERNATIONAL DE~CORR~ON ~E ET QES~SAUSSURE$
i t
~ ~
bullt
t t
UNIVERSIDAD PourtcNlCAVollncio7J1 NiwiemIif~ 1988
(SPAN
SECcl6lfn
BiOIogro marino Morine biologY ~1i1orine
Theunpub1ishedJllBIlu8criptEl have been reprod~d asr~ielved from the
authors The SCientiUc COlIIIBittee take not respoi~ulbll~~Y for any shy
error or omssi9n
Laa ponenciasno han side publicadas y estanreproducidas tal y como
sellan recibido demiddot BUSmiddot autorea pound1 Conte Cientfico middotno se responsemiddot
bil1za de cualquier error ~ om1si~n
bullbull _ bullbull_ bullbull ___ bull $_ _ L _ - _ --- -__
SESSION II Marine biology
Friday 11th November
SURFACE PROPERTIES ON NONmiddotTOXIC ANTIFOULING PAINT FILM
K Nanishi M Murase Y Yonehara M Kishihara and T Hirama
MICROFOULING ON ALUMINIUM PANELS PLACED IN THE TROPICAL WATERS
N B Bhosle K Venkat and A B Wagh
EFFECT OF TRIBUTYLTIN OXIDE (TBTO) ON THE OVARIAN MATURATION OF THE PRAWN CARIDINA WEBERI
R Nagabhushanam B Indira R Sarojini and Mary Frances Thompson
EFFECT OF BIOACTIVE MATERIALS FROM SOFT CORALS ON BACTERIA ASSOCIATED WITH COMMON FOULER
Vitalina Mary Avelin Mary R Sarojini and R Nagabhushanam
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
1
Inmiddot middotmiddot11middotmiddot f~~ a~ -tmiddot bull J~f
I~ W
i5iImiddot J~a t
ilmiddotmiddotmiddot I middotIt Ii
l
~
middotmiddot~~middotlL= g r
Lj Ic
t
i ~
I
gi II cEi ~ I I
1 t rW
D
tR bull
bull t 10
11
r
i
I ~
j
~
I
~bullbullbullbullbullbullbullbullbullbullbull
LV
FigS pH and p02 profiles into macrofoltling eomlllllllitilS dominated by Balm trenatU$ on steel plate 11 two pH profiles Ib) two lIOai profiles leI pH and P02 profiles into dead barllllCle shiella withdetri~Us I I
itIbl r p02 ~ I
nvnHg 11 70
j ifJi eo 1amp0
laquof it 30 s
I 20 N
ca 11 10 a o -- shy i bull
11
80
19
781 o 2 6 8 10 12 0 2 6 amiddot 10 12 dIpdI Qf ~iftto fouling In HighIst numbIII pIatIi
8deg~~
~i c5 iioo5 fiil
pH 8 bull_ ~
~~ ~
_ bullbullbull- pH
-pOz
(cJ
e-Clf ( j~JJlaquo1- --~I ~a ilil - _ I i ]1
~ ri en i ~ oJt ~iJ id tUI
2u1U I shyI t
shy1 2 3 6 6 7 8 9 10 11 12 i shyDepth 9f electrode (mm) into fouling
1 i
1
~----------~-------
~
~-~ lt1~
middotmiddotPmiddot ~
I
V
~
ill
1 Tbellitt~~ of cathodiC protection OO~~lIic~tlO aridrfOflllUlc
ofi AA~Uoullr14l ~1~I
I
ael~lner EG middotIUtcb8l1AJ Dept lt4 lInY1roMe1lal Bl01011
=~~venilt7 IlariCheW 1f139PL $ ~11al1df
r
I
Ablitrict ~~~ e~l billttI ~ted with ~pert1n ant11_1111ampp paint llere itpoii~1n _vatI- In the tl~ld iUIit 1bOioatbrt~ScM Were
1IUbjll~ to qtbocJ~C ~teltt1on byen _ current (~)SJi inci EDAX Iieamp~ta IJdlctecI ~ lachln teoc copper OIl tbePlCtec~ pa1n~ atter14 dQ1a~ protected Pllnt abo ebowed
) reduCed bacteiial activIty butmiddot ~e4 dltoe Iu1d protOzoanmiddot co~~l~tlont~1l IlU l~byenCOCltact not byen dUfwIl00 ~r _baRl lrillolvedln tOXln nil ereio noted
l ~~
1
I
Introduction
Applications of protective coatings and cathodic protect1on(cp~l
are recognised corrosion control methods for IIIBrine steel Itructureso Cathod~c protectlOll preventl conCIsion by Int~lng til8ctrl~l
currentS trem externalllClUfCa to e~~tthlJno~ e~ carrollon feetlans C04U lUChu tUauJ1ng Aelnts a
(
barrIer to ~ the flao of couos1a1 ~aboIHnu~r
Dccurrlng anodes cathodes or wlwit gelvWc CCIIII1 ( 19681
flinger 1980) AnUfauling ~ also JJICOqIOrII~ b1oc~~
Tr~ylt1n nuor~ (~) end o~ Oxide (~) (~ flJ1J
- of u blacld1111~
V _Ut Evans 1981) ~ ~
end growth of bipfoullng 0I1I)IIIII1IIIIe Wdd~ ca ~ of
paint rusendmiddotmanemiddot ~(r~ til )
As corrosian ~t1on ~ bICCIIIII -an ~1tlQd ~ of
antiroullnQs 1 conjunction _ith CP have bllClIOr ~
resultIng in reseudt 1rwestllj111tlng trw COIIIPIitlbUlty or VU~ ~t t _~
rOratlation to dlfrert levI ot CP (~Eelf 18110 ~
~t 1980) RelatIvely 1nIestiljlllUans have ~ urtw ~
or CP an thIt anUraullngpert~ of palnta
In thIt preseot 1nlIestioaUon tV~iln111 ronutlon ud
cOIIIHrclallY incanjlntlon wIth CP was ~t4IC~ Iw I sraed current (-966 l1li) arid exposed at the 1 Strait IInQlbullbullbully
~t1ons weremiddot 00 the IIhcIrt bnifects the PprIUd_
had an a) the various COIIIPQI1eIIts of n pe1ntf1l11 end b) l1oft~
developaentbull
bull bull ~
~ ~ - f
bull ~ _T~~-middot~ ~~- ~r _ -$1
(1) ~rlttionor Painted Steel Billets middot_ ~~tmiddot_tlJ middot~hmiddot~--~fmiddot ~ middot~A~-~~middot - -~ ~
Thirty-al ateel blllets (8-10aD2 end l depth) were cut frollbullt ~middot~middot~middotmiddot~middottrSmiddott ~jS-~ It bull gt~ tfl - middot--imiddotmiddot ~ t1dItrd steal plates The dimenSlonsof -thB butits-lim chosento
bull -~~ middotmiddotgt2h ~ ~-Imiddot ~~rt ~- -middoti~~ ni ~~middot1 l middot-~t~~middot _
(a) fecilltate amiddot Ie detrucUve -thxI of prePraUonpdor to ~~_gt gt~ k7i ~_ t~ 1--1- _ ) - +--_~ -_~bull ming electron ~ 4II1CI (iialloWdirectobs8iVatlCnS of the
- i lt-- ~ gtbull -o1e billet 8Uface
gt r~middotmiddotj ~L(~middot~middot rmiddot tmiddot J ~ - ~ - lt _ bull Single non-standard (1hIula~) nickel lIIireSmiddot Spot Welded e1Cing
bull ~1deJ ~ b~i~ ~j~~ tri afl~1b1ipOi~ f ~1t1 ~4middotmiddot f-gt -~ ~S-- ( -- - - _bull resin EactI billet then Undb~ tI1d prtiladlllltha5O __ eyilr
bull ~~_ ~~imiddot ~1iil h~ intlfcili~ ~eonbt1nlyi Tr1butylt1n F~id(18n)~~~O~dettu20i~blcicld8is 1IIfii bull i - ~ ( then 1IPI)l1ed to glvaadry Ulil tIi1c1ltnOss or 1(1)-150bullbull
tbull (2) IipreSSeCi Current Syat
bull A rigid ~tic due 1I8S drWedllliihwrtv-s41 liiIihOieS IIPProX~
bullbull 05-10 cia episrt ~ Pilnted ampUl8t Wsfbed IntaPQSIUon ampy
threadlng the nickel IIIlr8throu11 the dinto a tarproot junction
box Eliltilen of th8~1l1ii (recelving ~t-d curnritlere IIIlred
Into bull block connectOrrecelvlng dc trOll is potentiOstat The
rainJng iontroi blUets Wet8fbedfl11i1iiy lIIithln thejtlytlonbox
i~ityd~c IkiliPly ~~ ~lat1ruaWiode 8ncIacalOliel
refer~ electtodetoiere fllied ~ the per1aetat of the plut1cdlc IIIId
connected to the pot8ritlostat The desl~ of the potentlosbt ailoiolEld
~PlDteItlO1TClJrnnt to be MIllntained at 8prampset levtl of 966 II1II
c
r~
Grid raf 1J5s-t 114iH 563724 dud 0 Juna
Alolecule andbechr1a in general are ~ to
sutface lI1eroleyer (nlUSton) of _ter (~bullbull 191KJ)~To avold
contaJnatlonof ttw painted surf by U- -vents
blllets Were s-nact in a 1 conte1ner or atc1le (02i ) f~~nd
ter Iih1ch then f9ICI in tv 1IIhere the b~
be r-oved undeNeter El~~ bllleta fa Mcb trtai bullbullel
removed over a 1 dtq (336 hr~ expoeure perioda
each being ~~bn uposurnof 1 5 11 iA 168and DlIIauz1r
Arter field axpauHli the billets wan
fUtered _tar to ~ 1_ debtlflMd far 10 aJnuta ~
wv ~~~endattid sdbUiUd(otO
~ ~l f~ nn1ng el~~ --
lOOted onto 1middotCIi ~bullbullWIsend carbonmiddotcoa~nmiddotmiddot~middot f
each billet Ws~ Iis1ng a ccr1dge ~el~~
(5360) IIIlth anergy dJ8perslve X-ray alcroenelre1a (mx)
_lysis the les Wen reNed end coated III1th vold to al]Ql En
detailed sat of the alaquofa1l
(4) Lfiborattux ~
Painted taa~ bUletalllllre pnparad ~
in a Mrlne reclrculet1ng flOlll _ ~
f1Oll the ~ ~a1t~y Tha~~Un bullybullbullbull
10C M1nta1ned ~~t1as of ~t~la protazOllbull~bull10shy
(Entermap) ~ ~~~middotNlt~Eamp1~ __~and ~ r - ~ -
certaJn ~lc 1IQ1~ 1(1 the
the painted
~
thre repl1catft fn
plKed 1nto(022_)
- thIine1J~~ -lt bull
Atter mAX
(1-2) and i-n-t bn collected
14OC
Pain~
-
12 daY~ exposure and prepared as in d)~ - t
~I ~~~-- i~yen 1~ v
Rflltsi ~ _ ~f-~lI ft j bull _
rt~ll0~~~llY~~~~peln~stetsl~11i~ex~sed for
y~t1~J~ ~~~~1~1~~1UII~ya~kooJ~~~lt ~ ~~_~~I1J~ ~~~tomAX -lysis ~era than gold ~ bull bullbullbull c - _ bull lt
~~fllr ~~~ vllUll ~t1ans of the rfee htures and gt Co
~f~ - ~~~ ~f ~ ~aobta1Mdue IjIll811 iI1Flg 1~2 The IIIIdn ~~ ~ lt)ji9 )tmiddotmiddot - ht
- bull
~I rIIplt~ tIt8 -~ cOlllponanta of the pei1t 1bullbull WelIIlItr1x $ilj~t-~~middot - ~ _ -_
-IUch PVCbQed end rap~13y the chlorlna peak the
i~~~-~~-~V~~~-t18 ~~~~ ~~ Th8
ral~~i~ JlrQP(lrUo1sof ch 1Il-ntal paek~ll1d l~t_ Q~7 -gt- ~ _~ -- gt ) gt -_ -lt ~~ the 0 -end _ xprtsntabullTIwsa changes are -r1sed ~t~ lt~ _ ~ 1nT~ IThe8llia$lb npresentthe chengelI 1n~t1V8 p~lons
ti~~~ [ ~gt 3 - - - _ - _ -
Qf nellllllllt within We surface layars of the pe1ntmiddot ru The
~f~~thtJn -d ~rfeli avr the 336 h11cxI ampScanbe
SII4iIl r~ fig 3 al~ tht rta of~ dirfers tJetwaeo thaa ltmiddot~1- 1ft~- ~ _
IJOl appfiai-ed to ~1nlt1elly until ftar day 168 it fall
~~l~ 1h8relllllrelgn1fi~ diff~be~ qrOtactlid and
proieCted (at -966 ) piIIllnts tor lioo COPPer but not folimiddottin
Ibi~tlons Of the Pain -urface 8hoWad _Pil ~ witilt1M
(Plete I to IV)~ 1hePllat1na (~) pelnt(Phta x and b)
Clearl~st-i~~r alots on the SurfllC8 IiII1kh~ter1sed ~- f hlt1 lriitUl tin conCantrat1on~ lllariy slotsre Partially
~OIIe~ ~ tile _tr~tedal middot1lere rs ~an~ densely packed
grerltr~ ~th 1ar~(iIa1niy cotifIIr) 8ntt -ampniBil (lIB1rily 1rori)being
middot vl~l~~~~~ Dtt~i~~~t~ r~ ~f~~ I~t seawatllr for 11 n IIIOre slots appeared and th~ ~~ess _trb cwer
--if -- V~ c~~ f(i ~~~i~
(Plate Ie to It)~ the granules dld not protrude from the~rtace in ~ ~~~~j1~r~-ffJmiddot0~~
suhap~~r Both 0 nd --960 at ~~pa1nts ~ - fr ~~~~-w1-U~ ~~t
sl~llrat thi stag~ (Plates Ic-d II1d xrnspac~IlYh middotmiddotWith middot middotaJ~cmiddot gt incraallO ~alolre t~ difrrqcbullbull middotbatIiI8IImiddot the protected fjd
bull ~ ~ ~t~
IIllrotacbldpaint ~~lIfIt~ftitr~ h alQt~ p~t bull u bullbullbull J ~~i~~ 1~~
lIIith IIIuper lIdguon u 4lrotatad piIJnt (Pl8ti1 II) 1hD1II8S
Ill )1IMdw1lillS gnnJla~on f-r~~ dpert1cl ~ - - - ~~r ~t
Onthe 966 1IIpetnt t-var tIIO typeaofan with diffannt c t
~hIId~(Plate UI a to c) One hIId tIII-1 bull bullbull e _ bull
fUll wJ1hleas abvlaus alats the o1fwl bed C18UU1IlOt ~litUi middotC c ~ ~
less ~ t11 h Gnnulatlan __ but ~lIiIZly def1nlld in
middot~ latter Afte fila bed tiIIrtIId 10 ~are ~~j(
extansll(tlly ~ the~tIId(O at) tIIIIple lIIIdngthe~ - ~
middot orarallet1ai leas claaI (Plate I~) f u II1ao CQ1tJAJ11d to ~Gn - -
the -966 at 1 (Plata III d tor but 1JI8V8n 1nittNdcneabull -
Althatqa -middotslots wrs abvlaus becaI1ng 1 cwampr aiI- the orwWat1an8
f1nailyftar 14 day exposure ~ fUll drMIlapIIIent on all la - ~
are extensive with the 0 lhawlng both alota pore ~
OVIKgrOloln (Plata II c to f) Gnrulatlon 1bl to ~ gt~t~~ i
or the th1cknats of tho ma ~the ~ at ~ (~~~~ themiddot - - _ middott ~ ~ ~ - -1 1 t
middot f11bull IIIIIS IIOre extans1 thIIIn at dIlya but IIIIIIIY bull1Q~ tI st11l ~w -middotr~ 1gt ~1 ~
obvious fIIany ~ loI8re still p~ yen~~~~~~~r
than before Poiu appearedto be f~lf~ila~~c-~~~a (Plate IVf~ label 1) IIIIl1ch broke~i(2)~ ~~~~
t t
bull ~lS (3) ~ rflC~~ ~i~t~~l~~KI~ t~ldiIiiicyul~loIiih ~~Rht ~ )~- -~ - ~middotd 40 ( gt ~gt ~d lt1 t ~ Jt ~remiddot4amp tha paint trix cavity bottom eroded away reVe81~ng ore
t ~~t~ por~~ ~l~ P~]Y p~~8dby di~lutiaA~ ~~~~ti~Jr~gt~ta(l~~Siinll1cl1Ud~f iUiIiI reriatlGllbullt
bull a~~ ba~tid colCriAtloriccic~ GIl ~~~d ri -966 - ib~~~amp catQn1iti~~t~I~~(~t an ( ~
t the ~~~palnts (PIa_taiiI d~~h ~wh ~ n td have ~ CamplluJMitiIVc) ~~middottti o~lgtnalrf tlt of hIght
~ l=~A ~J~rW tYtles~ p~fiiilriQcOCCold to~Ja to jU-t ~jmiddoti~ai dlatClilS ~ir -jlati ~tbullbullent aNit ~ t ~~ th8middotf~ld~ies~~t~on~ ~ -966 prQ~~~(P14ta Iii~)Omiddotiliof~m1hiiHAiiipieat ere~ll-(piitir~ p-iot~iilans(PiA-iII~ end c) nbullbulltiledt dl~by1i~~ end wirinali1sheii ~cMI~bY thI tili
bull In t-~~ ~1i1nt 1oIIIeHfd~ ~m1IS (15OClind a t ri~r-dlatoa ~lor P~t dlataiOlonlt1anGIl ttw-966
t
bull ~
pr~t~ ~is~ih iion obitJauIJ A ~rot -=1 ~ ~ i __
reprasenttild on the 966 painted 1iUrfc1ncluicunljj ~ rwt
Achnanths Colonisation by bacteria 1n the laboretory bsed
bull eliperlMnt tollMct sbiUar pattern to the fild ~
bull bull Dilicueii
bull bull DUt 1Iidnt ~lona Mve dlttllrint ~~ ictian
It ~ JntM l1tenture ttt Iff yen __ JIll the
bull ~ bicins ani 111 ~t INchlnil (PhillIPs 1t73t EWene~ 1981
In ci1ntlHt tJWY ~tit ~Wl1c iIuCh as 1rbiltyl tirI
~ lite wAlII]ltiClr dlsP8tfu~ middotthiirsttilll~intaJn~ a
bull ~
lIIifoflll ampalCentraUon through the filII 10 thoIt biocide rel_ at tha
paint surfce 1fI$Jlts in IIIOre ditfusing ~s to repla~lj -iflJ~it
resulting 111 bullbulltNdy lr leechlng rata M1llat this IIIIY be middottruIi ot
tin ~ it was nOt true ~ ttw middotlBlf fOZWJlatiGn ua ~ bull - i - bull 1(1
ObSfvt1onundr SE clarly ~ tM t1l1 to be present
~ly lIS OIIIdle-llk parUc1~~t tn lUlfiice (Ito
IV) bUt usa ~ the dIpth attt fU-middot ~y~
If one considra the concn~rtlonmiddot chMge 111 1n ~111c COKICIl8rlts fro- the paint tU- i bullbullbull tMWei b~ ~ Si6t arid
tI-~t iron oxidt 1~ can bt (Fl_ 3 IrId TIibl8I) ~t ~ ase cona1derllble d1fftrenC8s ~ a Un decz bcdYMr ttw p1iod IIIIwraS lron~- an unt Ught inCr h ~ incqJper ~tlGn occUrnd _idly at ttw ~~ Gfb ~ period ~ out ttwn fu nPldly ampQII1n ~ umiddotirid II phast This IIIOUld bt tIlq)8Ctad It cantact l-=h1ng we occun1ng
IIltfIOS8d pertlcl of copper would lMch rapldly on t-n1on 1n - shy JIsewtr There would thlll1 ba bull J-a a1iltUy leu avalUba
i1 ~ ~
_tedal becfM uposed Thi tIOUld ttwn IIllCh Thla pulSing efttct
IIIOUld be expctad to -oth OIItlllith U- putic1 __ -lllible II tot 1~ GIl a on r-x _lao Thla teet 01 put1c1e 1tUng
can tit GIl the lIUrlace ClIlC8 it ~ a pan 1amp t
a 11aM bet in Plat I to IV IIaIrIg _ lit la_ II ~ 0- ~s _- PIate IW $llldl ~bull nshyot poRI flllNltttn Al~ tin ~ lou rete thtn I bullbullbull atill Ivwnce of bull al~t pulsed 1Itue rrlict tII1IIWd
elqlCtaG inIl ttwauUr ~ I I I
0
~ i
~ ~
~I J
bullbullt bullbullbullbullt bull bull ~
bull bull
I
bull t
~~
The same seqiHllle of cevity follliltloii (intttecaseofthetestjjltle
slot- shaped) fClliOileciby SecOlld8rYtlolesfotlllingin thll cavity
bottollis releasing further deep8rtox1ils (Plate XV d and g) As
cavities_re ProdICed ~ tiecaaIelnvadedby_ter~ onelilauld
expect 1ncreaa 1n1IOdha and dIladne lMls t the t1met
which the toxlM uw ~ Ths fCMld tCtHi eo (Flvmiddot 3)
~rttno the vi that_tat ~t1on otthe~ ~lY
il1Cl1l8S8S Idth UIiJ
Differences f~in toxin li~~to Ini 966 ill 8811P~ F ~ bOth Un end ~ lcMIr levels afillW
IIIIIlfI ptaInt(~1~~1ntha~~ ~~f~~ cst rklld with tbiIccpper ElJIc~l ta~Uona invalvinv
w pl8cId at a~ht~~middot3lO~lth Un cqJpel bull
(-960)10 ttW ~af this expei1tIIri1ts thIsiorelfJltdt -
that cqlp8r b8IiIId ~ IIiIOUldProc-ln f41ip1dly t thellllgllitiv
patentW _lied () as 1t 11 further f~1t ~ witt Un
III11ch is u-tat aqulllbd Idth
it If ttda 18 eo~ ~
~ )
frOll1 paintscould bl anqnced by an illprd current clithodic
protecUansysteipos1bly ahart_nino te 11fe of the paInts
antifouling ability Bacterial coloni Uon IIIU rtdUCed on the -s66 paint This
could have been due to unflSllOUlllble electrostatic cqrge on th surtece Th1a effect CIIl pley en IIIportant role in the t~of
bacteria to alrraces (lIIarsMll19E1O) As bacteria appear to IhiJw
greater toxicltyto copper (DMpsey 1981) it is Klre likely that
enhanced cqJper leacll1nl On the -966 AIIIPl c8used the recluCed
activity The IDlol8r toxicity ofUn to bacteria (as reported by
~ -
DelIIPsey Hle1gt or its lowes leaching rat was sUpPorted bythl
relatively rapid recolonisation of the slots by lIIicroorganiSlllS ~ftar thE inlt1al aurge of leachi~ Thailllpressecl current dto ~~ the oppoSite _ff~middot ~r on dtta colonstJon l1lI-1- in the
JIbcJrtory~ta larger rUDIIra f~ fit pro~td ~ther than uriprotected Pafnta
Ccnlus1one
1 tiild-a 18 presaIIUd ~ thattJn~ ~~
~llNChng notd1rf~ ~
2 TIleuseofCP~ ~CUIftnt(at) on1h1a ~fca41ng ~- ~
pa1I]t~ to ~_lAIIach1ngn_ at toxiN - -
3 leIJstaPidbectar1al ca1on1aatJon acaampmId an ~bId Pt1ntJ ~
4 JItore larger alcrofauJing (Oiat_ and PzOtoaoana) Gr~_
occurred on the protcbld~
I l
I
bull bull ~
mfEflpoundaS middot~-t9i _jgt- ~ _ _ ~sev M~J~ (1981) (cilonJzatiooof alUf~lng lnts by _rine
Jbull ~teda9g~1ta IIarina XXlV~ 185-191
- - gt 0 Ie 1 _ - _
poundV8I$i I~lf (1~P~h 1II8rin~Ugae and Foul lng AReviw ith
t p~l-i9Jrlr ~iiMdilto5hiP-fQUllfCiH 8ot~ ~r~ ~~~ XXIVbull lIPwl~J11A
Fotteth CN~R ~kk8nGB IWPIR1seJCiiiroaic1lihd ~e ~JIIth CII Offshore St1ucturu ElllS ~tjI~f~1Dt19i4
- - ~
UKC (1976) I~races ill ~W Ecology KIuverd Univbullbull middotraabullf~iddge ~bull C~~jII illt)Ia(t RC (1980) totinllsa Cattod1CPrOtect1cinbull middotmiddot8AtEmiddot~6haOCIlfif~ T~ p No 108 ~ 18 ~ 1IIHf bull
t bull lt ~J~~~ __ ~
Norkr4irulB(198D) Advanculn UcrOfj~lpound~ltiOyVol 4 (AlItxlaquoIder fl Ed) PlerIw Press lcxIdon~ lIP~ 51-85 ~ l~l - bull bull
PJijllt~i At(fm)~mr~~JnI~1fqul1nQ JlalntlJ Riumh ~JnOrpnic toatJngs 2 192middot
)~J gt0 c ~lt -- ~ __ RogerS~ Ut (1968) ina ~(lnt~fbItliP4ili GI COnoslon ~a Technology)bull bull
Sl~iorr~ VP~Ranald~ Rctd~~ (1S80)_ ~ Experllattal Stud1aiRelate middotEffect or Cathod1c Protict1Oi withmiddot terta1nr1c
Coating Syst~ 12th Annualofr T8Id1na1ogy Co1fItHnC8 liauston TIIlt p 9-12
j
fIG1 sPct FromMon IT8TJCuZOI Int Lliu EOAX
FlG~ Fro TlTCu20Ar-4 Dy Field Ellposult I~ir
il
I c
R hi -00amp=7
-I I I I I I I I I f
iii
bull bull
bullbullbull
I J
J
I Figur 3 Thl chan In reltl rqpoIQnlof -ariOtimiddotjOmponents of the paint film with time Thl vtic~1 ampkiS unl till _n ofmiddotmt Illment per unit OJ fe 50all Cl Na
Cu StI Feeshy
PlistinllFe S7~J6 704 3~74 908
f
r e ~ ee~
middotie ~lt - ~ shybullt bull ~
t -~ l Cu~ o ~ f
t --- 3igt - bull 0 J Sn
cc
t ~
~
bullo
~ bull e
bullbullbullbull bull ~
bull ebull a QI 1- bull 0 40 431282 316
101 dmllIII)bullt bullbullbullt
hr 0 ~~bulla 5~16 ~~ -~~ i166 4S i83 8~1B
24tu 6~480 SD~83
111middot 1~1 OS6
~ 5114 6~69 -51 UIamp middot016- lt ~rmiddot
1d o Mea 818 21 021
~Z21 199 1~1Qmiddot ~Jgt2 058
14 d o ~~ 804 Q~B8~~~a~~t~middot 685
866 372amp 8amp4 OS _i~11[ ~~64
Tttile 1 A81UvlL pzQJJortlcw 0 vxl~oI ~~bull Jn 1I1Dtitctel lt-1166) end lqIlDteCt6 (O)middot~af~_~middot_lodEDf~ middotmiddotmiddotmiddotmiddotmiddotmiddotmiddotmiddotc
~ ~
_----I~
J
I bull
I
J bull tmiddot
7o CONQRESOIMTERNACIONAL
I D~CORRO$IOr(IAAINAEmiddot~USTACIONES~v c)
t 7thflfERHATlONAL CONGRESS
ON IARINE CORROSION AND fO(JUNG
~CONGRE$INtERNATIONAL DE~CORR~ON ~E ET QES~SAUSSURE$
i t
~ ~
bullt
t t
UNIVERSIDAD PourtcNlCAVollncio7J1 NiwiemIif~ 1988
(SPAN
SECcl6lfn
BiOIogro marino Morine biologY ~1i1orine
Theunpub1ishedJllBIlu8criptEl have been reprod~d asr~ielved from the
authors The SCientiUc COlIIIBittee take not respoi~ulbll~~Y for any shy
error or omssi9n
Laa ponenciasno han side publicadas y estanreproducidas tal y como
sellan recibido demiddot BUSmiddot autorea pound1 Conte Cientfico middotno se responsemiddot
bil1za de cualquier error ~ om1si~n
bullbull _ bullbull_ bullbull ___ bull $_ _ L _ - _ --- -__
SESSION II Marine biology
Friday 11th November
SURFACE PROPERTIES ON NONmiddotTOXIC ANTIFOULING PAINT FILM
K Nanishi M Murase Y Yonehara M Kishihara and T Hirama
MICROFOULING ON ALUMINIUM PANELS PLACED IN THE TROPICAL WATERS
N B Bhosle K Venkat and A B Wagh
EFFECT OF TRIBUTYLTIN OXIDE (TBTO) ON THE OVARIAN MATURATION OF THE PRAWN CARIDINA WEBERI
R Nagabhushanam B Indira R Sarojini and Mary Frances Thompson
EFFECT OF BIOACTIVE MATERIALS FROM SOFT CORALS ON BACTERIA ASSOCIATED WITH COMMON FOULER
Vitalina Mary Avelin Mary R Sarojini and R Nagabhushanam
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
gi II cEi ~ I I
1 t rW
D
tR bull
bull t 10
11
r
i
I ~
j
~
I
~bullbullbullbullbullbullbullbullbullbullbull
LV
FigS pH and p02 profiles into macrofoltling eomlllllllitilS dominated by Balm trenatU$ on steel plate 11 two pH profiles Ib) two lIOai profiles leI pH and P02 profiles into dead barllllCle shiella withdetri~Us I I
itIbl r p02 ~ I
nvnHg 11 70
j ifJi eo 1amp0
laquof it 30 s
I 20 N
ca 11 10 a o -- shy i bull
11
80
19
781 o 2 6 8 10 12 0 2 6 amiddot 10 12 dIpdI Qf ~iftto fouling In HighIst numbIII pIatIi
8deg~~
~i c5 iioo5 fiil
pH 8 bull_ ~
~~ ~
_ bullbullbull- pH
-pOz
(cJ
e-Clf ( j~JJlaquo1- --~I ~a ilil - _ I i ]1
~ ri en i ~ oJt ~iJ id tUI
2u1U I shyI t
shy1 2 3 6 6 7 8 9 10 11 12 i shyDepth 9f electrode (mm) into fouling
1 i
1
~----------~-------
~
~-~ lt1~
middotmiddotPmiddot ~
I
V
~
ill
1 Tbellitt~~ of cathodiC protection OO~~lIic~tlO aridrfOflllUlc
ofi AA~Uoullr14l ~1~I
I
ael~lner EG middotIUtcb8l1AJ Dept lt4 lInY1roMe1lal Bl01011
=~~venilt7 IlariCheW 1f139PL $ ~11al1df
r
I
Ablitrict ~~~ e~l billttI ~ted with ~pert1n ant11_1111ampp paint llere itpoii~1n _vatI- In the tl~ld iUIit 1bOioatbrt~ScM Were
1IUbjll~ to qtbocJ~C ~teltt1on byen _ current (~)SJi inci EDAX Iieamp~ta IJdlctecI ~ lachln teoc copper OIl tbePlCtec~ pa1n~ atter14 dQ1a~ protected Pllnt abo ebowed
) reduCed bacteiial activIty butmiddot ~e4 dltoe Iu1d protOzoanmiddot co~~l~tlont~1l IlU l~byenCOCltact not byen dUfwIl00 ~r _baRl lrillolvedln tOXln nil ereio noted
l ~~
1
I
Introduction
Applications of protective coatings and cathodic protect1on(cp~l
are recognised corrosion control methods for IIIBrine steel Itructureso Cathod~c protectlOll preventl conCIsion by Int~lng til8ctrl~l
currentS trem externalllClUfCa to e~~tthlJno~ e~ carrollon feetlans C04U lUChu tUauJ1ng Aelnts a
(
barrIer to ~ the flao of couos1a1 ~aboIHnu~r
Dccurrlng anodes cathodes or wlwit gelvWc CCIIII1 ( 19681
flinger 1980) AnUfauling ~ also JJICOqIOrII~ b1oc~~
Tr~ylt1n nuor~ (~) end o~ Oxide (~) (~ flJ1J
- of u blacld1111~
V _Ut Evans 1981) ~ ~
end growth of bipfoullng 0I1I)IIIII1IIIIe Wdd~ ca ~ of
paint rusendmiddotmanemiddot ~(r~ til )
As corrosian ~t1on ~ bICCIIIII -an ~1tlQd ~ of
antiroullnQs 1 conjunction _ith CP have bllClIOr ~
resultIng in reseudt 1rwestllj111tlng trw COIIIPIitlbUlty or VU~ ~t t _~
rOratlation to dlfrert levI ot CP (~Eelf 18110 ~
~t 1980) RelatIvely 1nIestiljlllUans have ~ urtw ~
or CP an thIt anUraullngpert~ of palnta
In thIt preseot 1nlIestioaUon tV~iln111 ronutlon ud
cOIIIHrclallY incanjlntlon wIth CP was ~t4IC~ Iw I sraed current (-966 l1li) arid exposed at the 1 Strait IInQlbullbullbully
~t1ons weremiddot 00 the IIhcIrt bnifects the PprIUd_
had an a) the various COIIIPQI1eIIts of n pe1ntf1l11 end b) l1oft~
developaentbull
bull bull ~
~ ~ - f
bull ~ _T~~-middot~ ~~- ~r _ -$1
(1) ~rlttionor Painted Steel Billets middot_ ~~tmiddot_tlJ middot~hmiddot~--~fmiddot ~ middot~A~-~~middot - -~ ~
Thirty-al ateel blllets (8-10aD2 end l depth) were cut frollbullt ~middot~middot~middotmiddot~middottrSmiddott ~jS-~ It bull gt~ tfl - middot--imiddotmiddot ~ t1dItrd steal plates The dimenSlonsof -thB butits-lim chosento
bull -~~ middotmiddotgt2h ~ ~-Imiddot ~~rt ~- -middoti~~ ni ~~middot1 l middot-~t~~middot _
(a) fecilltate amiddot Ie detrucUve -thxI of prePraUonpdor to ~~_gt gt~ k7i ~_ t~ 1--1- _ ) - +--_~ -_~bull ming electron ~ 4II1CI (iialloWdirectobs8iVatlCnS of the
- i lt-- ~ gtbull -o1e billet 8Uface
gt r~middotmiddotj ~L(~middot~middot rmiddot tmiddot J ~ - ~ - lt _ bull Single non-standard (1hIula~) nickel lIIireSmiddot Spot Welded e1Cing
bull ~1deJ ~ b~i~ ~j~~ tri afl~1b1ipOi~ f ~1t1 ~4middotmiddot f-gt -~ ~S-- ( -- - - _bull resin EactI billet then Undb~ tI1d prtiladlllltha5O __ eyilr
bull ~~_ ~~imiddot ~1iil h~ intlfcili~ ~eonbt1nlyi Tr1butylt1n F~id(18n)~~~O~dettu20i~blcicld8is 1IIfii bull i - ~ ( then 1IPI)l1ed to glvaadry Ulil tIi1c1ltnOss or 1(1)-150bullbull
tbull (2) IipreSSeCi Current Syat
bull A rigid ~tic due 1I8S drWedllliihwrtv-s41 liiIihOieS IIPProX~
bullbull 05-10 cia episrt ~ Pilnted ampUl8t Wsfbed IntaPQSIUon ampy
threadlng the nickel IIIlr8throu11 the dinto a tarproot junction
box Eliltilen of th8~1l1ii (recelving ~t-d curnritlere IIIlred
Into bull block connectOrrecelvlng dc trOll is potentiOstat The
rainJng iontroi blUets Wet8fbedfl11i1iiy lIIithln thejtlytlonbox
i~ityd~c IkiliPly ~~ ~lat1ruaWiode 8ncIacalOliel
refer~ electtodetoiere fllied ~ the per1aetat of the plut1cdlc IIIId
connected to the pot8ritlostat The desl~ of the potentlosbt ailoiolEld
~PlDteItlO1TClJrnnt to be MIllntained at 8prampset levtl of 966 II1II
c
r~
Grid raf 1J5s-t 114iH 563724 dud 0 Juna
Alolecule andbechr1a in general are ~ to
sutface lI1eroleyer (nlUSton) of _ter (~bullbull 191KJ)~To avold
contaJnatlonof ttw painted surf by U- -vents
blllets Were s-nact in a 1 conte1ner or atc1le (02i ) f~~nd
ter Iih1ch then f9ICI in tv 1IIhere the b~
be r-oved undeNeter El~~ bllleta fa Mcb trtai bullbullel
removed over a 1 dtq (336 hr~ expoeure perioda
each being ~~bn uposurnof 1 5 11 iA 168and DlIIauz1r
Arter field axpauHli the billets wan
fUtered _tar to ~ 1_ debtlflMd far 10 aJnuta ~
wv ~~~endattid sdbUiUd(otO
~ ~l f~ nn1ng el~~ --
lOOted onto 1middotCIi ~bullbullWIsend carbonmiddotcoa~nmiddotmiddot~middot f
each billet Ws~ Iis1ng a ccr1dge ~el~~
(5360) IIIlth anergy dJ8perslve X-ray alcroenelre1a (mx)
_lysis the les Wen reNed end coated III1th vold to al]Ql En
detailed sat of the alaquofa1l
(4) Lfiborattux ~
Painted taa~ bUletalllllre pnparad ~
in a Mrlne reclrculet1ng flOlll _ ~
f1Oll the ~ ~a1t~y Tha~~Un bullybullbullbull
10C M1nta1ned ~~t1as of ~t~la protazOllbull~bull10shy
(Entermap) ~ ~~~middotNlt~Eamp1~ __~and ~ r - ~ -
certaJn ~lc 1IQ1~ 1(1 the
the painted
~
thre repl1catft fn
plKed 1nto(022_)
- thIine1J~~ -lt bull
Atter mAX
(1-2) and i-n-t bn collected
14OC
Pain~
-
12 daY~ exposure and prepared as in d)~ - t
~I ~~~-- i~yen 1~ v
Rflltsi ~ _ ~f-~lI ft j bull _
rt~ll0~~~llY~~~~peln~stetsl~11i~ex~sed for
y~t1~J~ ~~~~1~1~~1UII~ya~kooJ~~~lt ~ ~~_~~I1J~ ~~~tomAX -lysis ~era than gold ~ bull bullbullbull c - _ bull lt
~~fllr ~~~ vllUll ~t1ans of the rfee htures and gt Co
~f~ - ~~~ ~f ~ ~aobta1Mdue IjIll811 iI1Flg 1~2 The IIIIdn ~~ ~ lt)ji9 )tmiddotmiddot - ht
- bull
~I rIIplt~ tIt8 -~ cOlllponanta of the pei1t 1bullbull WelIIlItr1x $ilj~t-~~middot - ~ _ -_
-IUch PVCbQed end rap~13y the chlorlna peak the
i~~~-~~-~V~~~-t18 ~~~~ ~~ Th8
ral~~i~ JlrQP(lrUo1sof ch 1Il-ntal paek~ll1d l~t_ Q~7 -gt- ~ _~ -- gt ) gt -_ -lt ~~ the 0 -end _ xprtsntabullTIwsa changes are -r1sed ~t~ lt~ _ ~ 1nT~ IThe8llia$lb npresentthe chengelI 1n~t1V8 p~lons
ti~~~ [ ~gt 3 - - - _ - _ -
Qf nellllllllt within We surface layars of the pe1ntmiddot ru The
~f~~thtJn -d ~rfeli avr the 336 h11cxI ampScanbe
SII4iIl r~ fig 3 al~ tht rta of~ dirfers tJetwaeo thaa ltmiddot~1- 1ft~- ~ _
IJOl appfiai-ed to ~1nlt1elly until ftar day 168 it fall
~~l~ 1h8relllllrelgn1fi~ diff~be~ qrOtactlid and
proieCted (at -966 ) piIIllnts tor lioo COPPer but not folimiddottin
Ibi~tlons Of the Pain -urface 8hoWad _Pil ~ witilt1M
(Plete I to IV)~ 1hePllat1na (~) pelnt(Phta x and b)
Clearl~st-i~~r alots on the SurfllC8 IiII1kh~ter1sed ~- f hlt1 lriitUl tin conCantrat1on~ lllariy slotsre Partially
~OIIe~ ~ tile _tr~tedal middot1lere rs ~an~ densely packed
grerltr~ ~th 1ar~(iIa1niy cotifIIr) 8ntt -ampniBil (lIB1rily 1rori)being
middot vl~l~~~~~ Dtt~i~~~t~ r~ ~f~~ I~t seawatllr for 11 n IIIOre slots appeared and th~ ~~ess _trb cwer
--if -- V~ c~~ f(i ~~~i~
(Plate Ie to It)~ the granules dld not protrude from the~rtace in ~ ~~~~j1~r~-ffJmiddot0~~
suhap~~r Both 0 nd --960 at ~~pa1nts ~ - fr ~~~~-w1-U~ ~~t
sl~llrat thi stag~ (Plates Ic-d II1d xrnspac~IlYh middotmiddotWith middot middotaJ~cmiddot gt incraallO ~alolre t~ difrrqcbullbull middotbatIiI8IImiddot the protected fjd
bull ~ ~ ~t~
IIllrotacbldpaint ~~lIfIt~ftitr~ h alQt~ p~t bull u bullbullbull J ~~i~~ 1~~
lIIith IIIuper lIdguon u 4lrotatad piIJnt (Pl8ti1 II) 1hD1II8S
Ill )1IMdw1lillS gnnJla~on f-r~~ dpert1cl ~ - - - ~~r ~t
Onthe 966 1IIpetnt t-var tIIO typeaofan with diffannt c t
~hIId~(Plate UI a to c) One hIId tIII-1 bull bullbull e _ bull
fUll wJ1hleas abvlaus alats the o1fwl bed C18UU1IlOt ~litUi middotC c ~ ~
less ~ t11 h Gnnulatlan __ but ~lIiIZly def1nlld in
middot~ latter Afte fila bed tiIIrtIId 10 ~are ~~j(
extansll(tlly ~ the~tIId(O at) tIIIIple lIIIdngthe~ - ~
middot orarallet1ai leas claaI (Plate I~) f u II1ao CQ1tJAJ11d to ~Gn - -
the -966 at 1 (Plata III d tor but 1JI8V8n 1nittNdcneabull -
Althatqa -middotslots wrs abvlaus becaI1ng 1 cwampr aiI- the orwWat1an8
f1nailyftar 14 day exposure ~ fUll drMIlapIIIent on all la - ~
are extensive with the 0 lhawlng both alota pore ~
OVIKgrOloln (Plata II c to f) Gnrulatlon 1bl to ~ gt~t~~ i
or the th1cknats of tho ma ~the ~ at ~ (~~~~ themiddot - - _ middott ~ ~ ~ - -1 1 t
middot f11bull IIIIIS IIOre extans1 thIIIn at dIlya but IIIIIIIY bull1Q~ tI st11l ~w -middotr~ 1gt ~1 ~
obvious fIIany ~ loI8re still p~ yen~~~~~~~r
than before Poiu appearedto be f~lf~ila~~c-~~~a (Plate IVf~ label 1) IIIIl1ch broke~i(2)~ ~~~~
t t
bull ~lS (3) ~ rflC~~ ~i~t~~l~~KI~ t~ldiIiiicyul~loIiih ~~Rht ~ )~- -~ - ~middotd 40 ( gt ~gt ~d lt1 t ~ Jt ~remiddot4amp tha paint trix cavity bottom eroded away reVe81~ng ore
t ~~t~ por~~ ~l~ P~]Y p~~8dby di~lutiaA~ ~~~~ti~Jr~gt~ta(l~~Siinll1cl1Ud~f iUiIiI reriatlGllbullt
bull a~~ ba~tid colCriAtloriccic~ GIl ~~~d ri -966 - ib~~~amp catQn1iti~~t~I~~(~t an ( ~
t the ~~~palnts (PIa_taiiI d~~h ~wh ~ n td have ~ CamplluJMitiIVc) ~~middottti o~lgtnalrf tlt of hIght
~ l=~A ~J~rW tYtles~ p~fiiilriQcOCCold to~Ja to jU-t ~jmiddoti~ai dlatClilS ~ir -jlati ~tbullbullent aNit ~ t ~~ th8middotf~ld~ies~~t~on~ ~ -966 prQ~~~(P14ta Iii~)Omiddotiliof~m1hiiHAiiipieat ere~ll-(piitir~ p-iot~iilans(PiA-iII~ end c) nbullbulltiledt dl~by1i~~ end wirinali1sheii ~cMI~bY thI tili
bull In t-~~ ~1i1nt 1oIIIeHfd~ ~m1IS (15OClind a t ri~r-dlatoa ~lor P~t dlataiOlonlt1anGIl ttw-966
t
bull ~
pr~t~ ~is~ih iion obitJauIJ A ~rot -=1 ~ ~ i __
reprasenttild on the 966 painted 1iUrfc1ncluicunljj ~ rwt
Achnanths Colonisation by bacteria 1n the laboretory bsed
bull eliperlMnt tollMct sbiUar pattern to the fild ~
bull bull Dilicueii
bull bull DUt 1Iidnt ~lona Mve dlttllrint ~~ ictian
It ~ JntM l1tenture ttt Iff yen __ JIll the
bull ~ bicins ani 111 ~t INchlnil (PhillIPs 1t73t EWene~ 1981
In ci1ntlHt tJWY ~tit ~Wl1c iIuCh as 1rbiltyl tirI
~ lite wAlII]ltiClr dlsP8tfu~ middotthiirsttilll~intaJn~ a
bull ~
lIIifoflll ampalCentraUon through the filII 10 thoIt biocide rel_ at tha
paint surfce 1fI$Jlts in IIIOre ditfusing ~s to repla~lj -iflJ~it
resulting 111 bullbulltNdy lr leechlng rata M1llat this IIIIY be middottruIi ot
tin ~ it was nOt true ~ ttw middotlBlf fOZWJlatiGn ua ~ bull - i - bull 1(1
ObSfvt1onundr SE clarly ~ tM t1l1 to be present
~ly lIS OIIIdle-llk parUc1~~t tn lUlfiice (Ito
IV) bUt usa ~ the dIpth attt fU-middot ~y~
If one considra the concn~rtlonmiddot chMge 111 1n ~111c COKICIl8rlts fro- the paint tU- i bullbullbull tMWei b~ ~ Si6t arid
tI-~t iron oxidt 1~ can bt (Fl_ 3 IrId TIibl8I) ~t ~ ase cona1derllble d1fftrenC8s ~ a Un decz bcdYMr ttw p1iod IIIIwraS lron~- an unt Ught inCr h ~ incqJper ~tlGn occUrnd _idly at ttw ~~ Gfb ~ period ~ out ttwn fu nPldly ampQII1n ~ umiddotirid II phast This IIIOUld bt tIlq)8Ctad It cantact l-=h1ng we occun1ng
IIltfIOS8d pertlcl of copper would lMch rapldly on t-n1on 1n - shy JIsewtr There would thlll1 ba bull J-a a1iltUy leu avalUba
i1 ~ ~
_tedal becfM uposed Thi tIOUld ttwn IIllCh Thla pulSing efttct
IIIOUld be expctad to -oth OIItlllith U- putic1 __ -lllible II tot 1~ GIl a on r-x _lao Thla teet 01 put1c1e 1tUng
can tit GIl the lIUrlace ClIlC8 it ~ a pan 1amp t
a 11aM bet in Plat I to IV IIaIrIg _ lit la_ II ~ 0- ~s _- PIate IW $llldl ~bull nshyot poRI flllNltttn Al~ tin ~ lou rete thtn I bullbullbull atill Ivwnce of bull al~t pulsed 1Itue rrlict tII1IIWd
elqlCtaG inIl ttwauUr ~ I I I
0
~ i
~ ~
~I J
bullbullt bullbullbullbullt bull bull ~
bull bull
I
bull t
~~
The same seqiHllle of cevity follliltloii (intttecaseofthetestjjltle
slot- shaped) fClliOileciby SecOlld8rYtlolesfotlllingin thll cavity
bottollis releasing further deep8rtox1ils (Plate XV d and g) As
cavities_re ProdICed ~ tiecaaIelnvadedby_ter~ onelilauld
expect 1ncreaa 1n1IOdha and dIladne lMls t the t1met
which the toxlM uw ~ Ths fCMld tCtHi eo (Flvmiddot 3)
~rttno the vi that_tat ~t1on otthe~ ~lY
il1Cl1l8S8S Idth UIiJ
Differences f~in toxin li~~to Ini 966 ill 8811P~ F ~ bOth Un end ~ lcMIr levels afillW
IIIIIlfI ptaInt(~1~~1ntha~~ ~~f~~ cst rklld with tbiIccpper ElJIc~l ta~Uona invalvinv
w pl8cId at a~ht~~middot3lO~lth Un cqJpel bull
(-960)10 ttW ~af this expei1tIIri1ts thIsiorelfJltdt -
that cqlp8r b8IiIId ~ IIiIOUldProc-ln f41ip1dly t thellllgllitiv
patentW _lied () as 1t 11 further f~1t ~ witt Un
III11ch is u-tat aqulllbd Idth
it If ttda 18 eo~ ~
~ )
frOll1 paintscould bl anqnced by an illprd current clithodic
protecUansysteipos1bly ahart_nino te 11fe of the paInts
antifouling ability Bacterial coloni Uon IIIU rtdUCed on the -s66 paint This
could have been due to unflSllOUlllble electrostatic cqrge on th surtece Th1a effect CIIl pley en IIIportant role in the t~of
bacteria to alrraces (lIIarsMll19E1O) As bacteria appear to IhiJw
greater toxicltyto copper (DMpsey 1981) it is Klre likely that
enhanced cqJper leacll1nl On the -966 AIIIPl c8used the recluCed
activity The IDlol8r toxicity ofUn to bacteria (as reported by
~ -
DelIIPsey Hle1gt or its lowes leaching rat was sUpPorted bythl
relatively rapid recolonisation of the slots by lIIicroorganiSlllS ~ftar thE inlt1al aurge of leachi~ Thailllpressecl current dto ~~ the oppoSite _ff~middot ~r on dtta colonstJon l1lI-1- in the
JIbcJrtory~ta larger rUDIIra f~ fit pro~td ~ther than uriprotected Pafnta
Ccnlus1one
1 tiild-a 18 presaIIUd ~ thattJn~ ~~
~llNChng notd1rf~ ~
2 TIleuseofCP~ ~CUIftnt(at) on1h1a ~fca41ng ~- ~
pa1I]t~ to ~_lAIIach1ngn_ at toxiN - -
3 leIJstaPidbectar1al ca1on1aatJon acaampmId an ~bId Pt1ntJ ~
4 JItore larger alcrofauJing (Oiat_ and PzOtoaoana) Gr~_
occurred on the protcbld~
I l
I
bull bull ~
mfEflpoundaS middot~-t9i _jgt- ~ _ _ ~sev M~J~ (1981) (cilonJzatiooof alUf~lng lnts by _rine
Jbull ~teda9g~1ta IIarina XXlV~ 185-191
- - gt 0 Ie 1 _ - _
poundV8I$i I~lf (1~P~h 1II8rin~Ugae and Foul lng AReviw ith
t p~l-i9Jrlr ~iiMdilto5hiP-fQUllfCiH 8ot~ ~r~ ~~~ XXIVbull lIPwl~J11A
Fotteth CN~R ~kk8nGB IWPIR1seJCiiiroaic1lihd ~e ~JIIth CII Offshore St1ucturu ElllS ~tjI~f~1Dt19i4
- - ~
UKC (1976) I~races ill ~W Ecology KIuverd Univbullbull middotraabullf~iddge ~bull C~~jII illt)Ia(t RC (1980) totinllsa Cattod1CPrOtect1cinbull middotmiddot8AtEmiddot~6haOCIlfif~ T~ p No 108 ~ 18 ~ 1IIHf bull
t bull lt ~J~~~ __ ~
Norkr4irulB(198D) Advanculn UcrOfj~lpound~ltiOyVol 4 (AlItxlaquoIder fl Ed) PlerIw Press lcxIdon~ lIP~ 51-85 ~ l~l - bull bull
PJijllt~i At(fm)~mr~~JnI~1fqul1nQ JlalntlJ Riumh ~JnOrpnic toatJngs 2 192middot
)~J gt0 c ~lt -- ~ __ RogerS~ Ut (1968) ina ~(lnt~fbItliP4ili GI COnoslon ~a Technology)bull bull
Sl~iorr~ VP~Ranald~ Rctd~~ (1S80)_ ~ Experllattal Stud1aiRelate middotEffect or Cathod1c Protict1Oi withmiddot terta1nr1c
Coating Syst~ 12th Annualofr T8Id1na1ogy Co1fItHnC8 liauston TIIlt p 9-12
j
fIG1 sPct FromMon IT8TJCuZOI Int Lliu EOAX
FlG~ Fro TlTCu20Ar-4 Dy Field Ellposult I~ir
il
I c
R hi -00amp=7
-I I I I I I I I I f
iii
bull bull
bullbullbull
I J
J
I Figur 3 Thl chan In reltl rqpoIQnlof -ariOtimiddotjOmponents of the paint film with time Thl vtic~1 ampkiS unl till _n ofmiddotmt Illment per unit OJ fe 50all Cl Na
Cu StI Feeshy
PlistinllFe S7~J6 704 3~74 908
f
r e ~ ee~
middotie ~lt - ~ shybullt bull ~
t -~ l Cu~ o ~ f
t --- 3igt - bull 0 J Sn
cc
t ~
~
bullo
~ bull e
bullbullbullbull bull ~
bull ebull a QI 1- bull 0 40 431282 316
101 dmllIII)bullt bullbullbullt
hr 0 ~~bulla 5~16 ~~ -~~ i166 4S i83 8~1B
24tu 6~480 SD~83
111middot 1~1 OS6
~ 5114 6~69 -51 UIamp middot016- lt ~rmiddot
1d o Mea 818 21 021
~Z21 199 1~1Qmiddot ~Jgt2 058
14 d o ~~ 804 Q~B8~~~a~~t~middot 685
866 372amp 8amp4 OS _i~11[ ~~64
Tttile 1 A81UvlL pzQJJortlcw 0 vxl~oI ~~bull Jn 1I1Dtitctel lt-1166) end lqIlDteCt6 (O)middot~af~_~middot_lodEDf~ middotmiddotmiddotmiddotmiddotmiddotmiddotmiddotmiddotc
~ ~
_----I~
J
I bull
I
J bull tmiddot
7o CONQRESOIMTERNACIONAL
I D~CORRO$IOr(IAAINAEmiddot~USTACIONES~v c)
t 7thflfERHATlONAL CONGRESS
ON IARINE CORROSION AND fO(JUNG
~CONGRE$INtERNATIONAL DE~CORR~ON ~E ET QES~SAUSSURE$
i t
~ ~
bullt
t t
UNIVERSIDAD PourtcNlCAVollncio7J1 NiwiemIif~ 1988
(SPAN
SECcl6lfn
BiOIogro marino Morine biologY ~1i1orine
Theunpub1ishedJllBIlu8criptEl have been reprod~d asr~ielved from the
authors The SCientiUc COlIIIBittee take not respoi~ulbll~~Y for any shy
error or omssi9n
Laa ponenciasno han side publicadas y estanreproducidas tal y como
sellan recibido demiddot BUSmiddot autorea pound1 Conte Cientfico middotno se responsemiddot
bil1za de cualquier error ~ om1si~n
bullbull _ bullbull_ bullbull ___ bull $_ _ L _ - _ --- -__
SESSION II Marine biology
Friday 11th November
SURFACE PROPERTIES ON NONmiddotTOXIC ANTIFOULING PAINT FILM
K Nanishi M Murase Y Yonehara M Kishihara and T Hirama
MICROFOULING ON ALUMINIUM PANELS PLACED IN THE TROPICAL WATERS
N B Bhosle K Venkat and A B Wagh
EFFECT OF TRIBUTYLTIN OXIDE (TBTO) ON THE OVARIAN MATURATION OF THE PRAWN CARIDINA WEBERI
R Nagabhushanam B Indira R Sarojini and Mary Frances Thompson
EFFECT OF BIOACTIVE MATERIALS FROM SOFT CORALS ON BACTERIA ASSOCIATED WITH COMMON FOULER
Vitalina Mary Avelin Mary R Sarojini and R Nagabhushanam
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
LV
FigS pH and p02 profiles into macrofoltling eomlllllllitilS dominated by Balm trenatU$ on steel plate 11 two pH profiles Ib) two lIOai profiles leI pH and P02 profiles into dead barllllCle shiella withdetri~Us I I
itIbl r p02 ~ I
nvnHg 11 70
j ifJi eo 1amp0
laquof it 30 s
I 20 N
ca 11 10 a o -- shy i bull
11
80
19
781 o 2 6 8 10 12 0 2 6 amiddot 10 12 dIpdI Qf ~iftto fouling In HighIst numbIII pIatIi
8deg~~
~i c5 iioo5 fiil
pH 8 bull_ ~
~~ ~
_ bullbullbull- pH
-pOz
(cJ
e-Clf ( j~JJlaquo1- --~I ~a ilil - _ I i ]1
~ ri en i ~ oJt ~iJ id tUI
2u1U I shyI t
shy1 2 3 6 6 7 8 9 10 11 12 i shyDepth 9f electrode (mm) into fouling
1 i
1
~----------~-------
~
~-~ lt1~
middotmiddotPmiddot ~
I
V
~
ill
1 Tbellitt~~ of cathodiC protection OO~~lIic~tlO aridrfOflllUlc
ofi AA~Uoullr14l ~1~I
I
ael~lner EG middotIUtcb8l1AJ Dept lt4 lInY1roMe1lal Bl01011
=~~venilt7 IlariCheW 1f139PL $ ~11al1df
r
I
Ablitrict ~~~ e~l billttI ~ted with ~pert1n ant11_1111ampp paint llere itpoii~1n _vatI- In the tl~ld iUIit 1bOioatbrt~ScM Were
1IUbjll~ to qtbocJ~C ~teltt1on byen _ current (~)SJi inci EDAX Iieamp~ta IJdlctecI ~ lachln teoc copper OIl tbePlCtec~ pa1n~ atter14 dQ1a~ protected Pllnt abo ebowed
) reduCed bacteiial activIty butmiddot ~e4 dltoe Iu1d protOzoanmiddot co~~l~tlont~1l IlU l~byenCOCltact not byen dUfwIl00 ~r _baRl lrillolvedln tOXln nil ereio noted
l ~~
1
I
Introduction
Applications of protective coatings and cathodic protect1on(cp~l
are recognised corrosion control methods for IIIBrine steel Itructureso Cathod~c protectlOll preventl conCIsion by Int~lng til8ctrl~l
currentS trem externalllClUfCa to e~~tthlJno~ e~ carrollon feetlans C04U lUChu tUauJ1ng Aelnts a
(
barrIer to ~ the flao of couos1a1 ~aboIHnu~r
Dccurrlng anodes cathodes or wlwit gelvWc CCIIII1 ( 19681
flinger 1980) AnUfauling ~ also JJICOqIOrII~ b1oc~~
Tr~ylt1n nuor~ (~) end o~ Oxide (~) (~ flJ1J
- of u blacld1111~
V _Ut Evans 1981) ~ ~
end growth of bipfoullng 0I1I)IIIII1IIIIe Wdd~ ca ~ of
paint rusendmiddotmanemiddot ~(r~ til )
As corrosian ~t1on ~ bICCIIIII -an ~1tlQd ~ of
antiroullnQs 1 conjunction _ith CP have bllClIOr ~
resultIng in reseudt 1rwestllj111tlng trw COIIIPIitlbUlty or VU~ ~t t _~
rOratlation to dlfrert levI ot CP (~Eelf 18110 ~
~t 1980) RelatIvely 1nIestiljlllUans have ~ urtw ~
or CP an thIt anUraullngpert~ of palnta
In thIt preseot 1nlIestioaUon tV~iln111 ronutlon ud
cOIIIHrclallY incanjlntlon wIth CP was ~t4IC~ Iw I sraed current (-966 l1li) arid exposed at the 1 Strait IInQlbullbullbully
~t1ons weremiddot 00 the IIhcIrt bnifects the PprIUd_
had an a) the various COIIIPQI1eIIts of n pe1ntf1l11 end b) l1oft~
developaentbull
bull bull ~
~ ~ - f
bull ~ _T~~-middot~ ~~- ~r _ -$1
(1) ~rlttionor Painted Steel Billets middot_ ~~tmiddot_tlJ middot~hmiddot~--~fmiddot ~ middot~A~-~~middot - -~ ~
Thirty-al ateel blllets (8-10aD2 end l depth) were cut frollbullt ~middot~middot~middotmiddot~middottrSmiddott ~jS-~ It bull gt~ tfl - middot--imiddotmiddot ~ t1dItrd steal plates The dimenSlonsof -thB butits-lim chosento
bull -~~ middotmiddotgt2h ~ ~-Imiddot ~~rt ~- -middoti~~ ni ~~middot1 l middot-~t~~middot _
(a) fecilltate amiddot Ie detrucUve -thxI of prePraUonpdor to ~~_gt gt~ k7i ~_ t~ 1--1- _ ) - +--_~ -_~bull ming electron ~ 4II1CI (iialloWdirectobs8iVatlCnS of the
- i lt-- ~ gtbull -o1e billet 8Uface
gt r~middotmiddotj ~L(~middot~middot rmiddot tmiddot J ~ - ~ - lt _ bull Single non-standard (1hIula~) nickel lIIireSmiddot Spot Welded e1Cing
bull ~1deJ ~ b~i~ ~j~~ tri afl~1b1ipOi~ f ~1t1 ~4middotmiddot f-gt -~ ~S-- ( -- - - _bull resin EactI billet then Undb~ tI1d prtiladlllltha5O __ eyilr
bull ~~_ ~~imiddot ~1iil h~ intlfcili~ ~eonbt1nlyi Tr1butylt1n F~id(18n)~~~O~dettu20i~blcicld8is 1IIfii bull i - ~ ( then 1IPI)l1ed to glvaadry Ulil tIi1c1ltnOss or 1(1)-150bullbull
tbull (2) IipreSSeCi Current Syat
bull A rigid ~tic due 1I8S drWedllliihwrtv-s41 liiIihOieS IIPProX~
bullbull 05-10 cia episrt ~ Pilnted ampUl8t Wsfbed IntaPQSIUon ampy
threadlng the nickel IIIlr8throu11 the dinto a tarproot junction
box Eliltilen of th8~1l1ii (recelving ~t-d curnritlere IIIlred
Into bull block connectOrrecelvlng dc trOll is potentiOstat The
rainJng iontroi blUets Wet8fbedfl11i1iiy lIIithln thejtlytlonbox
i~ityd~c IkiliPly ~~ ~lat1ruaWiode 8ncIacalOliel
refer~ electtodetoiere fllied ~ the per1aetat of the plut1cdlc IIIId
connected to the pot8ritlostat The desl~ of the potentlosbt ailoiolEld
~PlDteItlO1TClJrnnt to be MIllntained at 8prampset levtl of 966 II1II
c
r~
Grid raf 1J5s-t 114iH 563724 dud 0 Juna
Alolecule andbechr1a in general are ~ to
sutface lI1eroleyer (nlUSton) of _ter (~bullbull 191KJ)~To avold
contaJnatlonof ttw painted surf by U- -vents
blllets Were s-nact in a 1 conte1ner or atc1le (02i ) f~~nd
ter Iih1ch then f9ICI in tv 1IIhere the b~
be r-oved undeNeter El~~ bllleta fa Mcb trtai bullbullel
removed over a 1 dtq (336 hr~ expoeure perioda
each being ~~bn uposurnof 1 5 11 iA 168and DlIIauz1r
Arter field axpauHli the billets wan
fUtered _tar to ~ 1_ debtlflMd far 10 aJnuta ~
wv ~~~endattid sdbUiUd(otO
~ ~l f~ nn1ng el~~ --
lOOted onto 1middotCIi ~bullbullWIsend carbonmiddotcoa~nmiddotmiddot~middot f
each billet Ws~ Iis1ng a ccr1dge ~el~~
(5360) IIIlth anergy dJ8perslve X-ray alcroenelre1a (mx)
_lysis the les Wen reNed end coated III1th vold to al]Ql En
detailed sat of the alaquofa1l
(4) Lfiborattux ~
Painted taa~ bUletalllllre pnparad ~
in a Mrlne reclrculet1ng flOlll _ ~
f1Oll the ~ ~a1t~y Tha~~Un bullybullbullbull
10C M1nta1ned ~~t1as of ~t~la protazOllbull~bull10shy
(Entermap) ~ ~~~middotNlt~Eamp1~ __~and ~ r - ~ -
certaJn ~lc 1IQ1~ 1(1 the
the painted
~
thre repl1catft fn
plKed 1nto(022_)
- thIine1J~~ -lt bull
Atter mAX
(1-2) and i-n-t bn collected
14OC
Pain~
-
12 daY~ exposure and prepared as in d)~ - t
~I ~~~-- i~yen 1~ v
Rflltsi ~ _ ~f-~lI ft j bull _
rt~ll0~~~llY~~~~peln~stetsl~11i~ex~sed for
y~t1~J~ ~~~~1~1~~1UII~ya~kooJ~~~lt ~ ~~_~~I1J~ ~~~tomAX -lysis ~era than gold ~ bull bullbullbull c - _ bull lt
~~fllr ~~~ vllUll ~t1ans of the rfee htures and gt Co
~f~ - ~~~ ~f ~ ~aobta1Mdue IjIll811 iI1Flg 1~2 The IIIIdn ~~ ~ lt)ji9 )tmiddotmiddot - ht
- bull
~I rIIplt~ tIt8 -~ cOlllponanta of the pei1t 1bullbull WelIIlItr1x $ilj~t-~~middot - ~ _ -_
-IUch PVCbQed end rap~13y the chlorlna peak the
i~~~-~~-~V~~~-t18 ~~~~ ~~ Th8
ral~~i~ JlrQP(lrUo1sof ch 1Il-ntal paek~ll1d l~t_ Q~7 -gt- ~ _~ -- gt ) gt -_ -lt ~~ the 0 -end _ xprtsntabullTIwsa changes are -r1sed ~t~ lt~ _ ~ 1nT~ IThe8llia$lb npresentthe chengelI 1n~t1V8 p~lons
ti~~~ [ ~gt 3 - - - _ - _ -
Qf nellllllllt within We surface layars of the pe1ntmiddot ru The
~f~~thtJn -d ~rfeli avr the 336 h11cxI ampScanbe
SII4iIl r~ fig 3 al~ tht rta of~ dirfers tJetwaeo thaa ltmiddot~1- 1ft~- ~ _
IJOl appfiai-ed to ~1nlt1elly until ftar day 168 it fall
~~l~ 1h8relllllrelgn1fi~ diff~be~ qrOtactlid and
proieCted (at -966 ) piIIllnts tor lioo COPPer but not folimiddottin
Ibi~tlons Of the Pain -urface 8hoWad _Pil ~ witilt1M
(Plete I to IV)~ 1hePllat1na (~) pelnt(Phta x and b)
Clearl~st-i~~r alots on the SurfllC8 IiII1kh~ter1sed ~- f hlt1 lriitUl tin conCantrat1on~ lllariy slotsre Partially
~OIIe~ ~ tile _tr~tedal middot1lere rs ~an~ densely packed
grerltr~ ~th 1ar~(iIa1niy cotifIIr) 8ntt -ampniBil (lIB1rily 1rori)being
middot vl~l~~~~~ Dtt~i~~~t~ r~ ~f~~ I~t seawatllr for 11 n IIIOre slots appeared and th~ ~~ess _trb cwer
--if -- V~ c~~ f(i ~~~i~
(Plate Ie to It)~ the granules dld not protrude from the~rtace in ~ ~~~~j1~r~-ffJmiddot0~~
suhap~~r Both 0 nd --960 at ~~pa1nts ~ - fr ~~~~-w1-U~ ~~t
sl~llrat thi stag~ (Plates Ic-d II1d xrnspac~IlYh middotmiddotWith middot middotaJ~cmiddot gt incraallO ~alolre t~ difrrqcbullbull middotbatIiI8IImiddot the protected fjd
bull ~ ~ ~t~
IIllrotacbldpaint ~~lIfIt~ftitr~ h alQt~ p~t bull u bullbullbull J ~~i~~ 1~~
lIIith IIIuper lIdguon u 4lrotatad piIJnt (Pl8ti1 II) 1hD1II8S
Ill )1IMdw1lillS gnnJla~on f-r~~ dpert1cl ~ - - - ~~r ~t
Onthe 966 1IIpetnt t-var tIIO typeaofan with diffannt c t
~hIId~(Plate UI a to c) One hIId tIII-1 bull bullbull e _ bull
fUll wJ1hleas abvlaus alats the o1fwl bed C18UU1IlOt ~litUi middotC c ~ ~
less ~ t11 h Gnnulatlan __ but ~lIiIZly def1nlld in
middot~ latter Afte fila bed tiIIrtIId 10 ~are ~~j(
extansll(tlly ~ the~tIId(O at) tIIIIple lIIIdngthe~ - ~
middot orarallet1ai leas claaI (Plate I~) f u II1ao CQ1tJAJ11d to ~Gn - -
the -966 at 1 (Plata III d tor but 1JI8V8n 1nittNdcneabull -
Althatqa -middotslots wrs abvlaus becaI1ng 1 cwampr aiI- the orwWat1an8
f1nailyftar 14 day exposure ~ fUll drMIlapIIIent on all la - ~
are extensive with the 0 lhawlng both alota pore ~
OVIKgrOloln (Plata II c to f) Gnrulatlon 1bl to ~ gt~t~~ i
or the th1cknats of tho ma ~the ~ at ~ (~~~~ themiddot - - _ middott ~ ~ ~ - -1 1 t
middot f11bull IIIIIS IIOre extans1 thIIIn at dIlya but IIIIIIIY bull1Q~ tI st11l ~w -middotr~ 1gt ~1 ~
obvious fIIany ~ loI8re still p~ yen~~~~~~~r
than before Poiu appearedto be f~lf~ila~~c-~~~a (Plate IVf~ label 1) IIIIl1ch broke~i(2)~ ~~~~
t t
bull ~lS (3) ~ rflC~~ ~i~t~~l~~KI~ t~ldiIiiicyul~loIiih ~~Rht ~ )~- -~ - ~middotd 40 ( gt ~gt ~d lt1 t ~ Jt ~remiddot4amp tha paint trix cavity bottom eroded away reVe81~ng ore
t ~~t~ por~~ ~l~ P~]Y p~~8dby di~lutiaA~ ~~~~ti~Jr~gt~ta(l~~Siinll1cl1Ud~f iUiIiI reriatlGllbullt
bull a~~ ba~tid colCriAtloriccic~ GIl ~~~d ri -966 - ib~~~amp catQn1iti~~t~I~~(~t an ( ~
t the ~~~palnts (PIa_taiiI d~~h ~wh ~ n td have ~ CamplluJMitiIVc) ~~middottti o~lgtnalrf tlt of hIght
~ l=~A ~J~rW tYtles~ p~fiiilriQcOCCold to~Ja to jU-t ~jmiddoti~ai dlatClilS ~ir -jlati ~tbullbullent aNit ~ t ~~ th8middotf~ld~ies~~t~on~ ~ -966 prQ~~~(P14ta Iii~)Omiddotiliof~m1hiiHAiiipieat ere~ll-(piitir~ p-iot~iilans(PiA-iII~ end c) nbullbulltiledt dl~by1i~~ end wirinali1sheii ~cMI~bY thI tili
bull In t-~~ ~1i1nt 1oIIIeHfd~ ~m1IS (15OClind a t ri~r-dlatoa ~lor P~t dlataiOlonlt1anGIl ttw-966
t
bull ~
pr~t~ ~is~ih iion obitJauIJ A ~rot -=1 ~ ~ i __
reprasenttild on the 966 painted 1iUrfc1ncluicunljj ~ rwt
Achnanths Colonisation by bacteria 1n the laboretory bsed
bull eliperlMnt tollMct sbiUar pattern to the fild ~
bull bull Dilicueii
bull bull DUt 1Iidnt ~lona Mve dlttllrint ~~ ictian
It ~ JntM l1tenture ttt Iff yen __ JIll the
bull ~ bicins ani 111 ~t INchlnil (PhillIPs 1t73t EWene~ 1981
In ci1ntlHt tJWY ~tit ~Wl1c iIuCh as 1rbiltyl tirI
~ lite wAlII]ltiClr dlsP8tfu~ middotthiirsttilll~intaJn~ a
bull ~
lIIifoflll ampalCentraUon through the filII 10 thoIt biocide rel_ at tha
paint surfce 1fI$Jlts in IIIOre ditfusing ~s to repla~lj -iflJ~it
resulting 111 bullbulltNdy lr leechlng rata M1llat this IIIIY be middottruIi ot
tin ~ it was nOt true ~ ttw middotlBlf fOZWJlatiGn ua ~ bull - i - bull 1(1
ObSfvt1onundr SE clarly ~ tM t1l1 to be present
~ly lIS OIIIdle-llk parUc1~~t tn lUlfiice (Ito
IV) bUt usa ~ the dIpth attt fU-middot ~y~
If one considra the concn~rtlonmiddot chMge 111 1n ~111c COKICIl8rlts fro- the paint tU- i bullbullbull tMWei b~ ~ Si6t arid
tI-~t iron oxidt 1~ can bt (Fl_ 3 IrId TIibl8I) ~t ~ ase cona1derllble d1fftrenC8s ~ a Un decz bcdYMr ttw p1iod IIIIwraS lron~- an unt Ught inCr h ~ incqJper ~tlGn occUrnd _idly at ttw ~~ Gfb ~ period ~ out ttwn fu nPldly ampQII1n ~ umiddotirid II phast This IIIOUld bt tIlq)8Ctad It cantact l-=h1ng we occun1ng
IIltfIOS8d pertlcl of copper would lMch rapldly on t-n1on 1n - shy JIsewtr There would thlll1 ba bull J-a a1iltUy leu avalUba
i1 ~ ~
_tedal becfM uposed Thi tIOUld ttwn IIllCh Thla pulSing efttct
IIIOUld be expctad to -oth OIItlllith U- putic1 __ -lllible II tot 1~ GIl a on r-x _lao Thla teet 01 put1c1e 1tUng
can tit GIl the lIUrlace ClIlC8 it ~ a pan 1amp t
a 11aM bet in Plat I to IV IIaIrIg _ lit la_ II ~ 0- ~s _- PIate IW $llldl ~bull nshyot poRI flllNltttn Al~ tin ~ lou rete thtn I bullbullbull atill Ivwnce of bull al~t pulsed 1Itue rrlict tII1IIWd
elqlCtaG inIl ttwauUr ~ I I I
0
~ i
~ ~
~I J
bullbullt bullbullbullbullt bull bull ~
bull bull
I
bull t
~~
The same seqiHllle of cevity follliltloii (intttecaseofthetestjjltle
slot- shaped) fClliOileciby SecOlld8rYtlolesfotlllingin thll cavity
bottollis releasing further deep8rtox1ils (Plate XV d and g) As
cavities_re ProdICed ~ tiecaaIelnvadedby_ter~ onelilauld
expect 1ncreaa 1n1IOdha and dIladne lMls t the t1met
which the toxlM uw ~ Ths fCMld tCtHi eo (Flvmiddot 3)
~rttno the vi that_tat ~t1on otthe~ ~lY
il1Cl1l8S8S Idth UIiJ
Differences f~in toxin li~~to Ini 966 ill 8811P~ F ~ bOth Un end ~ lcMIr levels afillW
IIIIIlfI ptaInt(~1~~1ntha~~ ~~f~~ cst rklld with tbiIccpper ElJIc~l ta~Uona invalvinv
w pl8cId at a~ht~~middot3lO~lth Un cqJpel bull
(-960)10 ttW ~af this expei1tIIri1ts thIsiorelfJltdt -
that cqlp8r b8IiIId ~ IIiIOUldProc-ln f41ip1dly t thellllgllitiv
patentW _lied () as 1t 11 further f~1t ~ witt Un
III11ch is u-tat aqulllbd Idth
it If ttda 18 eo~ ~
~ )
frOll1 paintscould bl anqnced by an illprd current clithodic
protecUansysteipos1bly ahart_nino te 11fe of the paInts
antifouling ability Bacterial coloni Uon IIIU rtdUCed on the -s66 paint This
could have been due to unflSllOUlllble electrostatic cqrge on th surtece Th1a effect CIIl pley en IIIportant role in the t~of
bacteria to alrraces (lIIarsMll19E1O) As bacteria appear to IhiJw
greater toxicltyto copper (DMpsey 1981) it is Klre likely that
enhanced cqJper leacll1nl On the -966 AIIIPl c8used the recluCed
activity The IDlol8r toxicity ofUn to bacteria (as reported by
~ -
DelIIPsey Hle1gt or its lowes leaching rat was sUpPorted bythl
relatively rapid recolonisation of the slots by lIIicroorganiSlllS ~ftar thE inlt1al aurge of leachi~ Thailllpressecl current dto ~~ the oppoSite _ff~middot ~r on dtta colonstJon l1lI-1- in the
JIbcJrtory~ta larger rUDIIra f~ fit pro~td ~ther than uriprotected Pafnta
Ccnlus1one
1 tiild-a 18 presaIIUd ~ thattJn~ ~~
~llNChng notd1rf~ ~
2 TIleuseofCP~ ~CUIftnt(at) on1h1a ~fca41ng ~- ~
pa1I]t~ to ~_lAIIach1ngn_ at toxiN - -
3 leIJstaPidbectar1al ca1on1aatJon acaampmId an ~bId Pt1ntJ ~
4 JItore larger alcrofauJing (Oiat_ and PzOtoaoana) Gr~_
occurred on the protcbld~
I l
I
bull bull ~
mfEflpoundaS middot~-t9i _jgt- ~ _ _ ~sev M~J~ (1981) (cilonJzatiooof alUf~lng lnts by _rine
Jbull ~teda9g~1ta IIarina XXlV~ 185-191
- - gt 0 Ie 1 _ - _
poundV8I$i I~lf (1~P~h 1II8rin~Ugae and Foul lng AReviw ith
t p~l-i9Jrlr ~iiMdilto5hiP-fQUllfCiH 8ot~ ~r~ ~~~ XXIVbull lIPwl~J11A
Fotteth CN~R ~kk8nGB IWPIR1seJCiiiroaic1lihd ~e ~JIIth CII Offshore St1ucturu ElllS ~tjI~f~1Dt19i4
- - ~
UKC (1976) I~races ill ~W Ecology KIuverd Univbullbull middotraabullf~iddge ~bull C~~jII illt)Ia(t RC (1980) totinllsa Cattod1CPrOtect1cinbull middotmiddot8AtEmiddot~6haOCIlfif~ T~ p No 108 ~ 18 ~ 1IIHf bull
t bull lt ~J~~~ __ ~
Norkr4irulB(198D) Advanculn UcrOfj~lpound~ltiOyVol 4 (AlItxlaquoIder fl Ed) PlerIw Press lcxIdon~ lIP~ 51-85 ~ l~l - bull bull
PJijllt~i At(fm)~mr~~JnI~1fqul1nQ JlalntlJ Riumh ~JnOrpnic toatJngs 2 192middot
)~J gt0 c ~lt -- ~ __ RogerS~ Ut (1968) ina ~(lnt~fbItliP4ili GI COnoslon ~a Technology)bull bull
Sl~iorr~ VP~Ranald~ Rctd~~ (1S80)_ ~ Experllattal Stud1aiRelate middotEffect or Cathod1c Protict1Oi withmiddot terta1nr1c
Coating Syst~ 12th Annualofr T8Id1na1ogy Co1fItHnC8 liauston TIIlt p 9-12
j
fIG1 sPct FromMon IT8TJCuZOI Int Lliu EOAX
FlG~ Fro TlTCu20Ar-4 Dy Field Ellposult I~ir
il
I c
R hi -00amp=7
-I I I I I I I I I f
iii
bull bull
bullbullbull
I J
J
I Figur 3 Thl chan In reltl rqpoIQnlof -ariOtimiddotjOmponents of the paint film with time Thl vtic~1 ampkiS unl till _n ofmiddotmt Illment per unit OJ fe 50all Cl Na
Cu StI Feeshy
PlistinllFe S7~J6 704 3~74 908
f
r e ~ ee~
middotie ~lt - ~ shybullt bull ~
t -~ l Cu~ o ~ f
t --- 3igt - bull 0 J Sn
cc
t ~
~
bullo
~ bull e
bullbullbullbull bull ~
bull ebull a QI 1- bull 0 40 431282 316
101 dmllIII)bullt bullbullbullt
hr 0 ~~bulla 5~16 ~~ -~~ i166 4S i83 8~1B
24tu 6~480 SD~83
111middot 1~1 OS6
~ 5114 6~69 -51 UIamp middot016- lt ~rmiddot
1d o Mea 818 21 021
~Z21 199 1~1Qmiddot ~Jgt2 058
14 d o ~~ 804 Q~B8~~~a~~t~middot 685
866 372amp 8amp4 OS _i~11[ ~~64
Tttile 1 A81UvlL pzQJJortlcw 0 vxl~oI ~~bull Jn 1I1Dtitctel lt-1166) end lqIlDteCt6 (O)middot~af~_~middot_lodEDf~ middotmiddotmiddotmiddotmiddotmiddotmiddotmiddotmiddotc
~ ~
_----I~
J
I bull
I
J bull tmiddot
7o CONQRESOIMTERNACIONAL
I D~CORRO$IOr(IAAINAEmiddot~USTACIONES~v c)
t 7thflfERHATlONAL CONGRESS
ON IARINE CORROSION AND fO(JUNG
~CONGRE$INtERNATIONAL DE~CORR~ON ~E ET QES~SAUSSURE$
i t
~ ~
bullt
t t
UNIVERSIDAD PourtcNlCAVollncio7J1 NiwiemIif~ 1988
(SPAN
SECcl6lfn
BiOIogro marino Morine biologY ~1i1orine
Theunpub1ishedJllBIlu8criptEl have been reprod~d asr~ielved from the
authors The SCientiUc COlIIIBittee take not respoi~ulbll~~Y for any shy
error or omssi9n
Laa ponenciasno han side publicadas y estanreproducidas tal y como
sellan recibido demiddot BUSmiddot autorea pound1 Conte Cientfico middotno se responsemiddot
bil1za de cualquier error ~ om1si~n
bullbull _ bullbull_ bullbull ___ bull $_ _ L _ - _ --- -__
SESSION II Marine biology
Friday 11th November
SURFACE PROPERTIES ON NONmiddotTOXIC ANTIFOULING PAINT FILM
K Nanishi M Murase Y Yonehara M Kishihara and T Hirama
MICROFOULING ON ALUMINIUM PANELS PLACED IN THE TROPICAL WATERS
N B Bhosle K Venkat and A B Wagh
EFFECT OF TRIBUTYLTIN OXIDE (TBTO) ON THE OVARIAN MATURATION OF THE PRAWN CARIDINA WEBERI
R Nagabhushanam B Indira R Sarojini and Mary Frances Thompson
EFFECT OF BIOACTIVE MATERIALS FROM SOFT CORALS ON BACTERIA ASSOCIATED WITH COMMON FOULER
Vitalina Mary Avelin Mary R Sarojini and R Nagabhushanam
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
~----------~-------
~
~-~ lt1~
middotmiddotPmiddot ~
I
V
~
ill
1 Tbellitt~~ of cathodiC protection OO~~lIic~tlO aridrfOflllUlc
ofi AA~Uoullr14l ~1~I
I
ael~lner EG middotIUtcb8l1AJ Dept lt4 lInY1roMe1lal Bl01011
=~~venilt7 IlariCheW 1f139PL $ ~11al1df
r
I
Ablitrict ~~~ e~l billttI ~ted with ~pert1n ant11_1111ampp paint llere itpoii~1n _vatI- In the tl~ld iUIit 1bOioatbrt~ScM Were
1IUbjll~ to qtbocJ~C ~teltt1on byen _ current (~)SJi inci EDAX Iieamp~ta IJdlctecI ~ lachln teoc copper OIl tbePlCtec~ pa1n~ atter14 dQ1a~ protected Pllnt abo ebowed
) reduCed bacteiial activIty butmiddot ~e4 dltoe Iu1d protOzoanmiddot co~~l~tlont~1l IlU l~byenCOCltact not byen dUfwIl00 ~r _baRl lrillolvedln tOXln nil ereio noted
l ~~
1
I
Introduction
Applications of protective coatings and cathodic protect1on(cp~l
are recognised corrosion control methods for IIIBrine steel Itructureso Cathod~c protectlOll preventl conCIsion by Int~lng til8ctrl~l
currentS trem externalllClUfCa to e~~tthlJno~ e~ carrollon feetlans C04U lUChu tUauJ1ng Aelnts a
(
barrIer to ~ the flao of couos1a1 ~aboIHnu~r
Dccurrlng anodes cathodes or wlwit gelvWc CCIIII1 ( 19681
flinger 1980) AnUfauling ~ also JJICOqIOrII~ b1oc~~
Tr~ylt1n nuor~ (~) end o~ Oxide (~) (~ flJ1J
- of u blacld1111~
V _Ut Evans 1981) ~ ~
end growth of bipfoullng 0I1I)IIIII1IIIIe Wdd~ ca ~ of
paint rusendmiddotmanemiddot ~(r~ til )
As corrosian ~t1on ~ bICCIIIII -an ~1tlQd ~ of
antiroullnQs 1 conjunction _ith CP have bllClIOr ~
resultIng in reseudt 1rwestllj111tlng trw COIIIPIitlbUlty or VU~ ~t t _~
rOratlation to dlfrert levI ot CP (~Eelf 18110 ~
~t 1980) RelatIvely 1nIestiljlllUans have ~ urtw ~
or CP an thIt anUraullngpert~ of palnta
In thIt preseot 1nlIestioaUon tV~iln111 ronutlon ud
cOIIIHrclallY incanjlntlon wIth CP was ~t4IC~ Iw I sraed current (-966 l1li) arid exposed at the 1 Strait IInQlbullbullbully
~t1ons weremiddot 00 the IIhcIrt bnifects the PprIUd_
had an a) the various COIIIPQI1eIIts of n pe1ntf1l11 end b) l1oft~
developaentbull
bull bull ~
~ ~ - f
bull ~ _T~~-middot~ ~~- ~r _ -$1
(1) ~rlttionor Painted Steel Billets middot_ ~~tmiddot_tlJ middot~hmiddot~--~fmiddot ~ middot~A~-~~middot - -~ ~
Thirty-al ateel blllets (8-10aD2 end l depth) were cut frollbullt ~middot~middot~middotmiddot~middottrSmiddott ~jS-~ It bull gt~ tfl - middot--imiddotmiddot ~ t1dItrd steal plates The dimenSlonsof -thB butits-lim chosento
bull -~~ middotmiddotgt2h ~ ~-Imiddot ~~rt ~- -middoti~~ ni ~~middot1 l middot-~t~~middot _
(a) fecilltate amiddot Ie detrucUve -thxI of prePraUonpdor to ~~_gt gt~ k7i ~_ t~ 1--1- _ ) - +--_~ -_~bull ming electron ~ 4II1CI (iialloWdirectobs8iVatlCnS of the
- i lt-- ~ gtbull -o1e billet 8Uface
gt r~middotmiddotj ~L(~middot~middot rmiddot tmiddot J ~ - ~ - lt _ bull Single non-standard (1hIula~) nickel lIIireSmiddot Spot Welded e1Cing
bull ~1deJ ~ b~i~ ~j~~ tri afl~1b1ipOi~ f ~1t1 ~4middotmiddot f-gt -~ ~S-- ( -- - - _bull resin EactI billet then Undb~ tI1d prtiladlllltha5O __ eyilr
bull ~~_ ~~imiddot ~1iil h~ intlfcili~ ~eonbt1nlyi Tr1butylt1n F~id(18n)~~~O~dettu20i~blcicld8is 1IIfii bull i - ~ ( then 1IPI)l1ed to glvaadry Ulil tIi1c1ltnOss or 1(1)-150bullbull
tbull (2) IipreSSeCi Current Syat
bull A rigid ~tic due 1I8S drWedllliihwrtv-s41 liiIihOieS IIPProX~
bullbull 05-10 cia episrt ~ Pilnted ampUl8t Wsfbed IntaPQSIUon ampy
threadlng the nickel IIIlr8throu11 the dinto a tarproot junction
box Eliltilen of th8~1l1ii (recelving ~t-d curnritlere IIIlred
Into bull block connectOrrecelvlng dc trOll is potentiOstat The
rainJng iontroi blUets Wet8fbedfl11i1iiy lIIithln thejtlytlonbox
i~ityd~c IkiliPly ~~ ~lat1ruaWiode 8ncIacalOliel
refer~ electtodetoiere fllied ~ the per1aetat of the plut1cdlc IIIId
connected to the pot8ritlostat The desl~ of the potentlosbt ailoiolEld
~PlDteItlO1TClJrnnt to be MIllntained at 8prampset levtl of 966 II1II
c
r~
Grid raf 1J5s-t 114iH 563724 dud 0 Juna
Alolecule andbechr1a in general are ~ to
sutface lI1eroleyer (nlUSton) of _ter (~bullbull 191KJ)~To avold
contaJnatlonof ttw painted surf by U- -vents
blllets Were s-nact in a 1 conte1ner or atc1le (02i ) f~~nd
ter Iih1ch then f9ICI in tv 1IIhere the b~
be r-oved undeNeter El~~ bllleta fa Mcb trtai bullbullel
removed over a 1 dtq (336 hr~ expoeure perioda
each being ~~bn uposurnof 1 5 11 iA 168and DlIIauz1r
Arter field axpauHli the billets wan
fUtered _tar to ~ 1_ debtlflMd far 10 aJnuta ~
wv ~~~endattid sdbUiUd(otO
~ ~l f~ nn1ng el~~ --
lOOted onto 1middotCIi ~bullbullWIsend carbonmiddotcoa~nmiddotmiddot~middot f
each billet Ws~ Iis1ng a ccr1dge ~el~~
(5360) IIIlth anergy dJ8perslve X-ray alcroenelre1a (mx)
_lysis the les Wen reNed end coated III1th vold to al]Ql En
detailed sat of the alaquofa1l
(4) Lfiborattux ~
Painted taa~ bUletalllllre pnparad ~
in a Mrlne reclrculet1ng flOlll _ ~
f1Oll the ~ ~a1t~y Tha~~Un bullybullbullbull
10C M1nta1ned ~~t1as of ~t~la protazOllbull~bull10shy
(Entermap) ~ ~~~middotNlt~Eamp1~ __~and ~ r - ~ -
certaJn ~lc 1IQ1~ 1(1 the
the painted
~
thre repl1catft fn
plKed 1nto(022_)
- thIine1J~~ -lt bull
Atter mAX
(1-2) and i-n-t bn collected
14OC
Pain~
-
12 daY~ exposure and prepared as in d)~ - t
~I ~~~-- i~yen 1~ v
Rflltsi ~ _ ~f-~lI ft j bull _
rt~ll0~~~llY~~~~peln~stetsl~11i~ex~sed for
y~t1~J~ ~~~~1~1~~1UII~ya~kooJ~~~lt ~ ~~_~~I1J~ ~~~tomAX -lysis ~era than gold ~ bull bullbullbull c - _ bull lt
~~fllr ~~~ vllUll ~t1ans of the rfee htures and gt Co
~f~ - ~~~ ~f ~ ~aobta1Mdue IjIll811 iI1Flg 1~2 The IIIIdn ~~ ~ lt)ji9 )tmiddotmiddot - ht
- bull
~I rIIplt~ tIt8 -~ cOlllponanta of the pei1t 1bullbull WelIIlItr1x $ilj~t-~~middot - ~ _ -_
-IUch PVCbQed end rap~13y the chlorlna peak the
i~~~-~~-~V~~~-t18 ~~~~ ~~ Th8
ral~~i~ JlrQP(lrUo1sof ch 1Il-ntal paek~ll1d l~t_ Q~7 -gt- ~ _~ -- gt ) gt -_ -lt ~~ the 0 -end _ xprtsntabullTIwsa changes are -r1sed ~t~ lt~ _ ~ 1nT~ IThe8llia$lb npresentthe chengelI 1n~t1V8 p~lons
ti~~~ [ ~gt 3 - - - _ - _ -
Qf nellllllllt within We surface layars of the pe1ntmiddot ru The
~f~~thtJn -d ~rfeli avr the 336 h11cxI ampScanbe
SII4iIl r~ fig 3 al~ tht rta of~ dirfers tJetwaeo thaa ltmiddot~1- 1ft~- ~ _
IJOl appfiai-ed to ~1nlt1elly until ftar day 168 it fall
~~l~ 1h8relllllrelgn1fi~ diff~be~ qrOtactlid and
proieCted (at -966 ) piIIllnts tor lioo COPPer but not folimiddottin
Ibi~tlons Of the Pain -urface 8hoWad _Pil ~ witilt1M
(Plete I to IV)~ 1hePllat1na (~) pelnt(Phta x and b)
Clearl~st-i~~r alots on the SurfllC8 IiII1kh~ter1sed ~- f hlt1 lriitUl tin conCantrat1on~ lllariy slotsre Partially
~OIIe~ ~ tile _tr~tedal middot1lere rs ~an~ densely packed
grerltr~ ~th 1ar~(iIa1niy cotifIIr) 8ntt -ampniBil (lIB1rily 1rori)being
middot vl~l~~~~~ Dtt~i~~~t~ r~ ~f~~ I~t seawatllr for 11 n IIIOre slots appeared and th~ ~~ess _trb cwer
--if -- V~ c~~ f(i ~~~i~
(Plate Ie to It)~ the granules dld not protrude from the~rtace in ~ ~~~~j1~r~-ffJmiddot0~~
suhap~~r Both 0 nd --960 at ~~pa1nts ~ - fr ~~~~-w1-U~ ~~t
sl~llrat thi stag~ (Plates Ic-d II1d xrnspac~IlYh middotmiddotWith middot middotaJ~cmiddot gt incraallO ~alolre t~ difrrqcbullbull middotbatIiI8IImiddot the protected fjd
bull ~ ~ ~t~
IIllrotacbldpaint ~~lIfIt~ftitr~ h alQt~ p~t bull u bullbullbull J ~~i~~ 1~~
lIIith IIIuper lIdguon u 4lrotatad piIJnt (Pl8ti1 II) 1hD1II8S
Ill )1IMdw1lillS gnnJla~on f-r~~ dpert1cl ~ - - - ~~r ~t
Onthe 966 1IIpetnt t-var tIIO typeaofan with diffannt c t
~hIId~(Plate UI a to c) One hIId tIII-1 bull bullbull e _ bull
fUll wJ1hleas abvlaus alats the o1fwl bed C18UU1IlOt ~litUi middotC c ~ ~
less ~ t11 h Gnnulatlan __ but ~lIiIZly def1nlld in
middot~ latter Afte fila bed tiIIrtIId 10 ~are ~~j(
extansll(tlly ~ the~tIId(O at) tIIIIple lIIIdngthe~ - ~
middot orarallet1ai leas claaI (Plate I~) f u II1ao CQ1tJAJ11d to ~Gn - -
the -966 at 1 (Plata III d tor but 1JI8V8n 1nittNdcneabull -
Althatqa -middotslots wrs abvlaus becaI1ng 1 cwampr aiI- the orwWat1an8
f1nailyftar 14 day exposure ~ fUll drMIlapIIIent on all la - ~
are extensive with the 0 lhawlng both alota pore ~
OVIKgrOloln (Plata II c to f) Gnrulatlon 1bl to ~ gt~t~~ i
or the th1cknats of tho ma ~the ~ at ~ (~~~~ themiddot - - _ middott ~ ~ ~ - -1 1 t
middot f11bull IIIIIS IIOre extans1 thIIIn at dIlya but IIIIIIIY bull1Q~ tI st11l ~w -middotr~ 1gt ~1 ~
obvious fIIany ~ loI8re still p~ yen~~~~~~~r
than before Poiu appearedto be f~lf~ila~~c-~~~a (Plate IVf~ label 1) IIIIl1ch broke~i(2)~ ~~~~
t t
bull ~lS (3) ~ rflC~~ ~i~t~~l~~KI~ t~ldiIiiicyul~loIiih ~~Rht ~ )~- -~ - ~middotd 40 ( gt ~gt ~d lt1 t ~ Jt ~remiddot4amp tha paint trix cavity bottom eroded away reVe81~ng ore
t ~~t~ por~~ ~l~ P~]Y p~~8dby di~lutiaA~ ~~~~ti~Jr~gt~ta(l~~Siinll1cl1Ud~f iUiIiI reriatlGllbullt
bull a~~ ba~tid colCriAtloriccic~ GIl ~~~d ri -966 - ib~~~amp catQn1iti~~t~I~~(~t an ( ~
t the ~~~palnts (PIa_taiiI d~~h ~wh ~ n td have ~ CamplluJMitiIVc) ~~middottti o~lgtnalrf tlt of hIght
~ l=~A ~J~rW tYtles~ p~fiiilriQcOCCold to~Ja to jU-t ~jmiddoti~ai dlatClilS ~ir -jlati ~tbullbullent aNit ~ t ~~ th8middotf~ld~ies~~t~on~ ~ -966 prQ~~~(P14ta Iii~)Omiddotiliof~m1hiiHAiiipieat ere~ll-(piitir~ p-iot~iilans(PiA-iII~ end c) nbullbulltiledt dl~by1i~~ end wirinali1sheii ~cMI~bY thI tili
bull In t-~~ ~1i1nt 1oIIIeHfd~ ~m1IS (15OClind a t ri~r-dlatoa ~lor P~t dlataiOlonlt1anGIl ttw-966
t
bull ~
pr~t~ ~is~ih iion obitJauIJ A ~rot -=1 ~ ~ i __
reprasenttild on the 966 painted 1iUrfc1ncluicunljj ~ rwt
Achnanths Colonisation by bacteria 1n the laboretory bsed
bull eliperlMnt tollMct sbiUar pattern to the fild ~
bull bull Dilicueii
bull bull DUt 1Iidnt ~lona Mve dlttllrint ~~ ictian
It ~ JntM l1tenture ttt Iff yen __ JIll the
bull ~ bicins ani 111 ~t INchlnil (PhillIPs 1t73t EWene~ 1981
In ci1ntlHt tJWY ~tit ~Wl1c iIuCh as 1rbiltyl tirI
~ lite wAlII]ltiClr dlsP8tfu~ middotthiirsttilll~intaJn~ a
bull ~
lIIifoflll ampalCentraUon through the filII 10 thoIt biocide rel_ at tha
paint surfce 1fI$Jlts in IIIOre ditfusing ~s to repla~lj -iflJ~it
resulting 111 bullbulltNdy lr leechlng rata M1llat this IIIIY be middottruIi ot
tin ~ it was nOt true ~ ttw middotlBlf fOZWJlatiGn ua ~ bull - i - bull 1(1
ObSfvt1onundr SE clarly ~ tM t1l1 to be present
~ly lIS OIIIdle-llk parUc1~~t tn lUlfiice (Ito
IV) bUt usa ~ the dIpth attt fU-middot ~y~
If one considra the concn~rtlonmiddot chMge 111 1n ~111c COKICIl8rlts fro- the paint tU- i bullbullbull tMWei b~ ~ Si6t arid
tI-~t iron oxidt 1~ can bt (Fl_ 3 IrId TIibl8I) ~t ~ ase cona1derllble d1fftrenC8s ~ a Un decz bcdYMr ttw p1iod IIIIwraS lron~- an unt Ught inCr h ~ incqJper ~tlGn occUrnd _idly at ttw ~~ Gfb ~ period ~ out ttwn fu nPldly ampQII1n ~ umiddotirid II phast This IIIOUld bt tIlq)8Ctad It cantact l-=h1ng we occun1ng
IIltfIOS8d pertlcl of copper would lMch rapldly on t-n1on 1n - shy JIsewtr There would thlll1 ba bull J-a a1iltUy leu avalUba
i1 ~ ~
_tedal becfM uposed Thi tIOUld ttwn IIllCh Thla pulSing efttct
IIIOUld be expctad to -oth OIItlllith U- putic1 __ -lllible II tot 1~ GIl a on r-x _lao Thla teet 01 put1c1e 1tUng
can tit GIl the lIUrlace ClIlC8 it ~ a pan 1amp t
a 11aM bet in Plat I to IV IIaIrIg _ lit la_ II ~ 0- ~s _- PIate IW $llldl ~bull nshyot poRI flllNltttn Al~ tin ~ lou rete thtn I bullbullbull atill Ivwnce of bull al~t pulsed 1Itue rrlict tII1IIWd
elqlCtaG inIl ttwauUr ~ I I I
0
~ i
~ ~
~I J
bullbullt bullbullbullbullt bull bull ~
bull bull
I
bull t
~~
The same seqiHllle of cevity follliltloii (intttecaseofthetestjjltle
slot- shaped) fClliOileciby SecOlld8rYtlolesfotlllingin thll cavity
bottollis releasing further deep8rtox1ils (Plate XV d and g) As
cavities_re ProdICed ~ tiecaaIelnvadedby_ter~ onelilauld
expect 1ncreaa 1n1IOdha and dIladne lMls t the t1met
which the toxlM uw ~ Ths fCMld tCtHi eo (Flvmiddot 3)
~rttno the vi that_tat ~t1on otthe~ ~lY
il1Cl1l8S8S Idth UIiJ
Differences f~in toxin li~~to Ini 966 ill 8811P~ F ~ bOth Un end ~ lcMIr levels afillW
IIIIIlfI ptaInt(~1~~1ntha~~ ~~f~~ cst rklld with tbiIccpper ElJIc~l ta~Uona invalvinv
w pl8cId at a~ht~~middot3lO~lth Un cqJpel bull
(-960)10 ttW ~af this expei1tIIri1ts thIsiorelfJltdt -
that cqlp8r b8IiIId ~ IIiIOUldProc-ln f41ip1dly t thellllgllitiv
patentW _lied () as 1t 11 further f~1t ~ witt Un
III11ch is u-tat aqulllbd Idth
it If ttda 18 eo~ ~
~ )
frOll1 paintscould bl anqnced by an illprd current clithodic
protecUansysteipos1bly ahart_nino te 11fe of the paInts
antifouling ability Bacterial coloni Uon IIIU rtdUCed on the -s66 paint This
could have been due to unflSllOUlllble electrostatic cqrge on th surtece Th1a effect CIIl pley en IIIportant role in the t~of
bacteria to alrraces (lIIarsMll19E1O) As bacteria appear to IhiJw
greater toxicltyto copper (DMpsey 1981) it is Klre likely that
enhanced cqJper leacll1nl On the -966 AIIIPl c8used the recluCed
activity The IDlol8r toxicity ofUn to bacteria (as reported by
~ -
DelIIPsey Hle1gt or its lowes leaching rat was sUpPorted bythl
relatively rapid recolonisation of the slots by lIIicroorganiSlllS ~ftar thE inlt1al aurge of leachi~ Thailllpressecl current dto ~~ the oppoSite _ff~middot ~r on dtta colonstJon l1lI-1- in the
JIbcJrtory~ta larger rUDIIra f~ fit pro~td ~ther than uriprotected Pafnta
Ccnlus1one
1 tiild-a 18 presaIIUd ~ thattJn~ ~~
~llNChng notd1rf~ ~
2 TIleuseofCP~ ~CUIftnt(at) on1h1a ~fca41ng ~- ~
pa1I]t~ to ~_lAIIach1ngn_ at toxiN - -
3 leIJstaPidbectar1al ca1on1aatJon acaampmId an ~bId Pt1ntJ ~
4 JItore larger alcrofauJing (Oiat_ and PzOtoaoana) Gr~_
occurred on the protcbld~
I l
I
bull bull ~
mfEflpoundaS middot~-t9i _jgt- ~ _ _ ~sev M~J~ (1981) (cilonJzatiooof alUf~lng lnts by _rine
Jbull ~teda9g~1ta IIarina XXlV~ 185-191
- - gt 0 Ie 1 _ - _
poundV8I$i I~lf (1~P~h 1II8rin~Ugae and Foul lng AReviw ith
t p~l-i9Jrlr ~iiMdilto5hiP-fQUllfCiH 8ot~ ~r~ ~~~ XXIVbull lIPwl~J11A
Fotteth CN~R ~kk8nGB IWPIR1seJCiiiroaic1lihd ~e ~JIIth CII Offshore St1ucturu ElllS ~tjI~f~1Dt19i4
- - ~
UKC (1976) I~races ill ~W Ecology KIuverd Univbullbull middotraabullf~iddge ~bull C~~jII illt)Ia(t RC (1980) totinllsa Cattod1CPrOtect1cinbull middotmiddot8AtEmiddot~6haOCIlfif~ T~ p No 108 ~ 18 ~ 1IIHf bull
t bull lt ~J~~~ __ ~
Norkr4irulB(198D) Advanculn UcrOfj~lpound~ltiOyVol 4 (AlItxlaquoIder fl Ed) PlerIw Press lcxIdon~ lIP~ 51-85 ~ l~l - bull bull
PJijllt~i At(fm)~mr~~JnI~1fqul1nQ JlalntlJ Riumh ~JnOrpnic toatJngs 2 192middot
)~J gt0 c ~lt -- ~ __ RogerS~ Ut (1968) ina ~(lnt~fbItliP4ili GI COnoslon ~a Technology)bull bull
Sl~iorr~ VP~Ranald~ Rctd~~ (1S80)_ ~ Experllattal Stud1aiRelate middotEffect or Cathod1c Protict1Oi withmiddot terta1nr1c
Coating Syst~ 12th Annualofr T8Id1na1ogy Co1fItHnC8 liauston TIIlt p 9-12
j
fIG1 sPct FromMon IT8TJCuZOI Int Lliu EOAX
FlG~ Fro TlTCu20Ar-4 Dy Field Ellposult I~ir
il
I c
R hi -00amp=7
-I I I I I I I I I f
iii
bull bull
bullbullbull
I J
J
I Figur 3 Thl chan In reltl rqpoIQnlof -ariOtimiddotjOmponents of the paint film with time Thl vtic~1 ampkiS unl till _n ofmiddotmt Illment per unit OJ fe 50all Cl Na
Cu StI Feeshy
PlistinllFe S7~J6 704 3~74 908
f
r e ~ ee~
middotie ~lt - ~ shybullt bull ~
t -~ l Cu~ o ~ f
t --- 3igt - bull 0 J Sn
cc
t ~
~
bullo
~ bull e
bullbullbullbull bull ~
bull ebull a QI 1- bull 0 40 431282 316
101 dmllIII)bullt bullbullbullt
hr 0 ~~bulla 5~16 ~~ -~~ i166 4S i83 8~1B
24tu 6~480 SD~83
111middot 1~1 OS6
~ 5114 6~69 -51 UIamp middot016- lt ~rmiddot
1d o Mea 818 21 021
~Z21 199 1~1Qmiddot ~Jgt2 058
14 d o ~~ 804 Q~B8~~~a~~t~middot 685
866 372amp 8amp4 OS _i~11[ ~~64
Tttile 1 A81UvlL pzQJJortlcw 0 vxl~oI ~~bull Jn 1I1Dtitctel lt-1166) end lqIlDteCt6 (O)middot~af~_~middot_lodEDf~ middotmiddotmiddotmiddotmiddotmiddotmiddotmiddotmiddotc
~ ~
_----I~
J
I bull
I
J bull tmiddot
7o CONQRESOIMTERNACIONAL
I D~CORRO$IOr(IAAINAEmiddot~USTACIONES~v c)
t 7thflfERHATlONAL CONGRESS
ON IARINE CORROSION AND fO(JUNG
~CONGRE$INtERNATIONAL DE~CORR~ON ~E ET QES~SAUSSURE$
i t
~ ~
bullt
t t
UNIVERSIDAD PourtcNlCAVollncio7J1 NiwiemIif~ 1988
(SPAN
SECcl6lfn
BiOIogro marino Morine biologY ~1i1orine
Theunpub1ishedJllBIlu8criptEl have been reprod~d asr~ielved from the
authors The SCientiUc COlIIIBittee take not respoi~ulbll~~Y for any shy
error or omssi9n
Laa ponenciasno han side publicadas y estanreproducidas tal y como
sellan recibido demiddot BUSmiddot autorea pound1 Conte Cientfico middotno se responsemiddot
bil1za de cualquier error ~ om1si~n
bullbull _ bullbull_ bullbull ___ bull $_ _ L _ - _ --- -__
SESSION II Marine biology
Friday 11th November
SURFACE PROPERTIES ON NONmiddotTOXIC ANTIFOULING PAINT FILM
K Nanishi M Murase Y Yonehara M Kishihara and T Hirama
MICROFOULING ON ALUMINIUM PANELS PLACED IN THE TROPICAL WATERS
N B Bhosle K Venkat and A B Wagh
EFFECT OF TRIBUTYLTIN OXIDE (TBTO) ON THE OVARIAN MATURATION OF THE PRAWN CARIDINA WEBERI
R Nagabhushanam B Indira R Sarojini and Mary Frances Thompson
EFFECT OF BIOACTIVE MATERIALS FROM SOFT CORALS ON BACTERIA ASSOCIATED WITH COMMON FOULER
Vitalina Mary Avelin Mary R Sarojini and R Nagabhushanam
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
I
V
~
ill
1 Tbellitt~~ of cathodiC protection OO~~lIic~tlO aridrfOflllUlc
ofi AA~Uoullr14l ~1~I
I
ael~lner EG middotIUtcb8l1AJ Dept lt4 lInY1roMe1lal Bl01011
=~~venilt7 IlariCheW 1f139PL $ ~11al1df
r
I
Ablitrict ~~~ e~l billttI ~ted with ~pert1n ant11_1111ampp paint llere itpoii~1n _vatI- In the tl~ld iUIit 1bOioatbrt~ScM Were
1IUbjll~ to qtbocJ~C ~teltt1on byen _ current (~)SJi inci EDAX Iieamp~ta IJdlctecI ~ lachln teoc copper OIl tbePlCtec~ pa1n~ atter14 dQ1a~ protected Pllnt abo ebowed
) reduCed bacteiial activIty butmiddot ~e4 dltoe Iu1d protOzoanmiddot co~~l~tlont~1l IlU l~byenCOCltact not byen dUfwIl00 ~r _baRl lrillolvedln tOXln nil ereio noted
l ~~
1
I
Introduction
Applications of protective coatings and cathodic protect1on(cp~l
are recognised corrosion control methods for IIIBrine steel Itructureso Cathod~c protectlOll preventl conCIsion by Int~lng til8ctrl~l
currentS trem externalllClUfCa to e~~tthlJno~ e~ carrollon feetlans C04U lUChu tUauJ1ng Aelnts a
(
barrIer to ~ the flao of couos1a1 ~aboIHnu~r
Dccurrlng anodes cathodes or wlwit gelvWc CCIIII1 ( 19681
flinger 1980) AnUfauling ~ also JJICOqIOrII~ b1oc~~
Tr~ylt1n nuor~ (~) end o~ Oxide (~) (~ flJ1J
- of u blacld1111~
V _Ut Evans 1981) ~ ~
end growth of bipfoullng 0I1I)IIIII1IIIIe Wdd~ ca ~ of
paint rusendmiddotmanemiddot ~(r~ til )
As corrosian ~t1on ~ bICCIIIII -an ~1tlQd ~ of
antiroullnQs 1 conjunction _ith CP have bllClIOr ~
resultIng in reseudt 1rwestllj111tlng trw COIIIPIitlbUlty or VU~ ~t t _~
rOratlation to dlfrert levI ot CP (~Eelf 18110 ~
~t 1980) RelatIvely 1nIestiljlllUans have ~ urtw ~
or CP an thIt anUraullngpert~ of palnta
In thIt preseot 1nlIestioaUon tV~iln111 ronutlon ud
cOIIIHrclallY incanjlntlon wIth CP was ~t4IC~ Iw I sraed current (-966 l1li) arid exposed at the 1 Strait IInQlbullbullbully
~t1ons weremiddot 00 the IIhcIrt bnifects the PprIUd_
had an a) the various COIIIPQI1eIIts of n pe1ntf1l11 end b) l1oft~
developaentbull
bull bull ~
~ ~ - f
bull ~ _T~~-middot~ ~~- ~r _ -$1
(1) ~rlttionor Painted Steel Billets middot_ ~~tmiddot_tlJ middot~hmiddot~--~fmiddot ~ middot~A~-~~middot - -~ ~
Thirty-al ateel blllets (8-10aD2 end l depth) were cut frollbullt ~middot~middot~middotmiddot~middottrSmiddott ~jS-~ It bull gt~ tfl - middot--imiddotmiddot ~ t1dItrd steal plates The dimenSlonsof -thB butits-lim chosento
bull -~~ middotmiddotgt2h ~ ~-Imiddot ~~rt ~- -middoti~~ ni ~~middot1 l middot-~t~~middot _
(a) fecilltate amiddot Ie detrucUve -thxI of prePraUonpdor to ~~_gt gt~ k7i ~_ t~ 1--1- _ ) - +--_~ -_~bull ming electron ~ 4II1CI (iialloWdirectobs8iVatlCnS of the
- i lt-- ~ gtbull -o1e billet 8Uface
gt r~middotmiddotj ~L(~middot~middot rmiddot tmiddot J ~ - ~ - lt _ bull Single non-standard (1hIula~) nickel lIIireSmiddot Spot Welded e1Cing
bull ~1deJ ~ b~i~ ~j~~ tri afl~1b1ipOi~ f ~1t1 ~4middotmiddot f-gt -~ ~S-- ( -- - - _bull resin EactI billet then Undb~ tI1d prtiladlllltha5O __ eyilr
bull ~~_ ~~imiddot ~1iil h~ intlfcili~ ~eonbt1nlyi Tr1butylt1n F~id(18n)~~~O~dettu20i~blcicld8is 1IIfii bull i - ~ ( then 1IPI)l1ed to glvaadry Ulil tIi1c1ltnOss or 1(1)-150bullbull
tbull (2) IipreSSeCi Current Syat
bull A rigid ~tic due 1I8S drWedllliihwrtv-s41 liiIihOieS IIPProX~
bullbull 05-10 cia episrt ~ Pilnted ampUl8t Wsfbed IntaPQSIUon ampy
threadlng the nickel IIIlr8throu11 the dinto a tarproot junction
box Eliltilen of th8~1l1ii (recelving ~t-d curnritlere IIIlred
Into bull block connectOrrecelvlng dc trOll is potentiOstat The
rainJng iontroi blUets Wet8fbedfl11i1iiy lIIithln thejtlytlonbox
i~ityd~c IkiliPly ~~ ~lat1ruaWiode 8ncIacalOliel
refer~ electtodetoiere fllied ~ the per1aetat of the plut1cdlc IIIId
connected to the pot8ritlostat The desl~ of the potentlosbt ailoiolEld
~PlDteItlO1TClJrnnt to be MIllntained at 8prampset levtl of 966 II1II
c
r~
Grid raf 1J5s-t 114iH 563724 dud 0 Juna
Alolecule andbechr1a in general are ~ to
sutface lI1eroleyer (nlUSton) of _ter (~bullbull 191KJ)~To avold
contaJnatlonof ttw painted surf by U- -vents
blllets Were s-nact in a 1 conte1ner or atc1le (02i ) f~~nd
ter Iih1ch then f9ICI in tv 1IIhere the b~
be r-oved undeNeter El~~ bllleta fa Mcb trtai bullbullel
removed over a 1 dtq (336 hr~ expoeure perioda
each being ~~bn uposurnof 1 5 11 iA 168and DlIIauz1r
Arter field axpauHli the billets wan
fUtered _tar to ~ 1_ debtlflMd far 10 aJnuta ~
wv ~~~endattid sdbUiUd(otO
~ ~l f~ nn1ng el~~ --
lOOted onto 1middotCIi ~bullbullWIsend carbonmiddotcoa~nmiddotmiddot~middot f
each billet Ws~ Iis1ng a ccr1dge ~el~~
(5360) IIIlth anergy dJ8perslve X-ray alcroenelre1a (mx)
_lysis the les Wen reNed end coated III1th vold to al]Ql En
detailed sat of the alaquofa1l
(4) Lfiborattux ~
Painted taa~ bUletalllllre pnparad ~
in a Mrlne reclrculet1ng flOlll _ ~
f1Oll the ~ ~a1t~y Tha~~Un bullybullbullbull
10C M1nta1ned ~~t1as of ~t~la protazOllbull~bull10shy
(Entermap) ~ ~~~middotNlt~Eamp1~ __~and ~ r - ~ -
certaJn ~lc 1IQ1~ 1(1 the
the painted
~
thre repl1catft fn
plKed 1nto(022_)
- thIine1J~~ -lt bull
Atter mAX
(1-2) and i-n-t bn collected
14OC
Pain~
-
12 daY~ exposure and prepared as in d)~ - t
~I ~~~-- i~yen 1~ v
Rflltsi ~ _ ~f-~lI ft j bull _
rt~ll0~~~llY~~~~peln~stetsl~11i~ex~sed for
y~t1~J~ ~~~~1~1~~1UII~ya~kooJ~~~lt ~ ~~_~~I1J~ ~~~tomAX -lysis ~era than gold ~ bull bullbullbull c - _ bull lt
~~fllr ~~~ vllUll ~t1ans of the rfee htures and gt Co
~f~ - ~~~ ~f ~ ~aobta1Mdue IjIll811 iI1Flg 1~2 The IIIIdn ~~ ~ lt)ji9 )tmiddotmiddot - ht
- bull
~I rIIplt~ tIt8 -~ cOlllponanta of the pei1t 1bullbull WelIIlItr1x $ilj~t-~~middot - ~ _ -_
-IUch PVCbQed end rap~13y the chlorlna peak the
i~~~-~~-~V~~~-t18 ~~~~ ~~ Th8
ral~~i~ JlrQP(lrUo1sof ch 1Il-ntal paek~ll1d l~t_ Q~7 -gt- ~ _~ -- gt ) gt -_ -lt ~~ the 0 -end _ xprtsntabullTIwsa changes are -r1sed ~t~ lt~ _ ~ 1nT~ IThe8llia$lb npresentthe chengelI 1n~t1V8 p~lons
ti~~~ [ ~gt 3 - - - _ - _ -
Qf nellllllllt within We surface layars of the pe1ntmiddot ru The
~f~~thtJn -d ~rfeli avr the 336 h11cxI ampScanbe
SII4iIl r~ fig 3 al~ tht rta of~ dirfers tJetwaeo thaa ltmiddot~1- 1ft~- ~ _
IJOl appfiai-ed to ~1nlt1elly until ftar day 168 it fall
~~l~ 1h8relllllrelgn1fi~ diff~be~ qrOtactlid and
proieCted (at -966 ) piIIllnts tor lioo COPPer but not folimiddottin
Ibi~tlons Of the Pain -urface 8hoWad _Pil ~ witilt1M
(Plete I to IV)~ 1hePllat1na (~) pelnt(Phta x and b)
Clearl~st-i~~r alots on the SurfllC8 IiII1kh~ter1sed ~- f hlt1 lriitUl tin conCantrat1on~ lllariy slotsre Partially
~OIIe~ ~ tile _tr~tedal middot1lere rs ~an~ densely packed
grerltr~ ~th 1ar~(iIa1niy cotifIIr) 8ntt -ampniBil (lIB1rily 1rori)being
middot vl~l~~~~~ Dtt~i~~~t~ r~ ~f~~ I~t seawatllr for 11 n IIIOre slots appeared and th~ ~~ess _trb cwer
--if -- V~ c~~ f(i ~~~i~
(Plate Ie to It)~ the granules dld not protrude from the~rtace in ~ ~~~~j1~r~-ffJmiddot0~~
suhap~~r Both 0 nd --960 at ~~pa1nts ~ - fr ~~~~-w1-U~ ~~t
sl~llrat thi stag~ (Plates Ic-d II1d xrnspac~IlYh middotmiddotWith middot middotaJ~cmiddot gt incraallO ~alolre t~ difrrqcbullbull middotbatIiI8IImiddot the protected fjd
bull ~ ~ ~t~
IIllrotacbldpaint ~~lIfIt~ftitr~ h alQt~ p~t bull u bullbullbull J ~~i~~ 1~~
lIIith IIIuper lIdguon u 4lrotatad piIJnt (Pl8ti1 II) 1hD1II8S
Ill )1IMdw1lillS gnnJla~on f-r~~ dpert1cl ~ - - - ~~r ~t
Onthe 966 1IIpetnt t-var tIIO typeaofan with diffannt c t
~hIId~(Plate UI a to c) One hIId tIII-1 bull bullbull e _ bull
fUll wJ1hleas abvlaus alats the o1fwl bed C18UU1IlOt ~litUi middotC c ~ ~
less ~ t11 h Gnnulatlan __ but ~lIiIZly def1nlld in
middot~ latter Afte fila bed tiIIrtIId 10 ~are ~~j(
extansll(tlly ~ the~tIId(O at) tIIIIple lIIIdngthe~ - ~
middot orarallet1ai leas claaI (Plate I~) f u II1ao CQ1tJAJ11d to ~Gn - -
the -966 at 1 (Plata III d tor but 1JI8V8n 1nittNdcneabull -
Althatqa -middotslots wrs abvlaus becaI1ng 1 cwampr aiI- the orwWat1an8
f1nailyftar 14 day exposure ~ fUll drMIlapIIIent on all la - ~
are extensive with the 0 lhawlng both alota pore ~
OVIKgrOloln (Plata II c to f) Gnrulatlon 1bl to ~ gt~t~~ i
or the th1cknats of tho ma ~the ~ at ~ (~~~~ themiddot - - _ middott ~ ~ ~ - -1 1 t
middot f11bull IIIIIS IIOre extans1 thIIIn at dIlya but IIIIIIIY bull1Q~ tI st11l ~w -middotr~ 1gt ~1 ~
obvious fIIany ~ loI8re still p~ yen~~~~~~~r
than before Poiu appearedto be f~lf~ila~~c-~~~a (Plate IVf~ label 1) IIIIl1ch broke~i(2)~ ~~~~
t t
bull ~lS (3) ~ rflC~~ ~i~t~~l~~KI~ t~ldiIiiicyul~loIiih ~~Rht ~ )~- -~ - ~middotd 40 ( gt ~gt ~d lt1 t ~ Jt ~remiddot4amp tha paint trix cavity bottom eroded away reVe81~ng ore
t ~~t~ por~~ ~l~ P~]Y p~~8dby di~lutiaA~ ~~~~ti~Jr~gt~ta(l~~Siinll1cl1Ud~f iUiIiI reriatlGllbullt
bull a~~ ba~tid colCriAtloriccic~ GIl ~~~d ri -966 - ib~~~amp catQn1iti~~t~I~~(~t an ( ~
t the ~~~palnts (PIa_taiiI d~~h ~wh ~ n td have ~ CamplluJMitiIVc) ~~middottti o~lgtnalrf tlt of hIght
~ l=~A ~J~rW tYtles~ p~fiiilriQcOCCold to~Ja to jU-t ~jmiddoti~ai dlatClilS ~ir -jlati ~tbullbullent aNit ~ t ~~ th8middotf~ld~ies~~t~on~ ~ -966 prQ~~~(P14ta Iii~)Omiddotiliof~m1hiiHAiiipieat ere~ll-(piitir~ p-iot~iilans(PiA-iII~ end c) nbullbulltiledt dl~by1i~~ end wirinali1sheii ~cMI~bY thI tili
bull In t-~~ ~1i1nt 1oIIIeHfd~ ~m1IS (15OClind a t ri~r-dlatoa ~lor P~t dlataiOlonlt1anGIl ttw-966
t
bull ~
pr~t~ ~is~ih iion obitJauIJ A ~rot -=1 ~ ~ i __
reprasenttild on the 966 painted 1iUrfc1ncluicunljj ~ rwt
Achnanths Colonisation by bacteria 1n the laboretory bsed
bull eliperlMnt tollMct sbiUar pattern to the fild ~
bull bull Dilicueii
bull bull DUt 1Iidnt ~lona Mve dlttllrint ~~ ictian
It ~ JntM l1tenture ttt Iff yen __ JIll the
bull ~ bicins ani 111 ~t INchlnil (PhillIPs 1t73t EWene~ 1981
In ci1ntlHt tJWY ~tit ~Wl1c iIuCh as 1rbiltyl tirI
~ lite wAlII]ltiClr dlsP8tfu~ middotthiirsttilll~intaJn~ a
bull ~
lIIifoflll ampalCentraUon through the filII 10 thoIt biocide rel_ at tha
paint surfce 1fI$Jlts in IIIOre ditfusing ~s to repla~lj -iflJ~it
resulting 111 bullbulltNdy lr leechlng rata M1llat this IIIIY be middottruIi ot
tin ~ it was nOt true ~ ttw middotlBlf fOZWJlatiGn ua ~ bull - i - bull 1(1
ObSfvt1onundr SE clarly ~ tM t1l1 to be present
~ly lIS OIIIdle-llk parUc1~~t tn lUlfiice (Ito
IV) bUt usa ~ the dIpth attt fU-middot ~y~
If one considra the concn~rtlonmiddot chMge 111 1n ~111c COKICIl8rlts fro- the paint tU- i bullbullbull tMWei b~ ~ Si6t arid
tI-~t iron oxidt 1~ can bt (Fl_ 3 IrId TIibl8I) ~t ~ ase cona1derllble d1fftrenC8s ~ a Un decz bcdYMr ttw p1iod IIIIwraS lron~- an unt Ught inCr h ~ incqJper ~tlGn occUrnd _idly at ttw ~~ Gfb ~ period ~ out ttwn fu nPldly ampQII1n ~ umiddotirid II phast This IIIOUld bt tIlq)8Ctad It cantact l-=h1ng we occun1ng
IIltfIOS8d pertlcl of copper would lMch rapldly on t-n1on 1n - shy JIsewtr There would thlll1 ba bull J-a a1iltUy leu avalUba
i1 ~ ~
_tedal becfM uposed Thi tIOUld ttwn IIllCh Thla pulSing efttct
IIIOUld be expctad to -oth OIItlllith U- putic1 __ -lllible II tot 1~ GIl a on r-x _lao Thla teet 01 put1c1e 1tUng
can tit GIl the lIUrlace ClIlC8 it ~ a pan 1amp t
a 11aM bet in Plat I to IV IIaIrIg _ lit la_ II ~ 0- ~s _- PIate IW $llldl ~bull nshyot poRI flllNltttn Al~ tin ~ lou rete thtn I bullbullbull atill Ivwnce of bull al~t pulsed 1Itue rrlict tII1IIWd
elqlCtaG inIl ttwauUr ~ I I I
0
~ i
~ ~
~I J
bullbullt bullbullbullbullt bull bull ~
bull bull
I
bull t
~~
The same seqiHllle of cevity follliltloii (intttecaseofthetestjjltle
slot- shaped) fClliOileciby SecOlld8rYtlolesfotlllingin thll cavity
bottollis releasing further deep8rtox1ils (Plate XV d and g) As
cavities_re ProdICed ~ tiecaaIelnvadedby_ter~ onelilauld
expect 1ncreaa 1n1IOdha and dIladne lMls t the t1met
which the toxlM uw ~ Ths fCMld tCtHi eo (Flvmiddot 3)
~rttno the vi that_tat ~t1on otthe~ ~lY
il1Cl1l8S8S Idth UIiJ
Differences f~in toxin li~~to Ini 966 ill 8811P~ F ~ bOth Un end ~ lcMIr levels afillW
IIIIIlfI ptaInt(~1~~1ntha~~ ~~f~~ cst rklld with tbiIccpper ElJIc~l ta~Uona invalvinv
w pl8cId at a~ht~~middot3lO~lth Un cqJpel bull
(-960)10 ttW ~af this expei1tIIri1ts thIsiorelfJltdt -
that cqlp8r b8IiIId ~ IIiIOUldProc-ln f41ip1dly t thellllgllitiv
patentW _lied () as 1t 11 further f~1t ~ witt Un
III11ch is u-tat aqulllbd Idth
it If ttda 18 eo~ ~
~ )
frOll1 paintscould bl anqnced by an illprd current clithodic
protecUansysteipos1bly ahart_nino te 11fe of the paInts
antifouling ability Bacterial coloni Uon IIIU rtdUCed on the -s66 paint This
could have been due to unflSllOUlllble electrostatic cqrge on th surtece Th1a effect CIIl pley en IIIportant role in the t~of
bacteria to alrraces (lIIarsMll19E1O) As bacteria appear to IhiJw
greater toxicltyto copper (DMpsey 1981) it is Klre likely that
enhanced cqJper leacll1nl On the -966 AIIIPl c8used the recluCed
activity The IDlol8r toxicity ofUn to bacteria (as reported by
~ -
DelIIPsey Hle1gt or its lowes leaching rat was sUpPorted bythl
relatively rapid recolonisation of the slots by lIIicroorganiSlllS ~ftar thE inlt1al aurge of leachi~ Thailllpressecl current dto ~~ the oppoSite _ff~middot ~r on dtta colonstJon l1lI-1- in the
JIbcJrtory~ta larger rUDIIra f~ fit pro~td ~ther than uriprotected Pafnta
Ccnlus1one
1 tiild-a 18 presaIIUd ~ thattJn~ ~~
~llNChng notd1rf~ ~
2 TIleuseofCP~ ~CUIftnt(at) on1h1a ~fca41ng ~- ~
pa1I]t~ to ~_lAIIach1ngn_ at toxiN - -
3 leIJstaPidbectar1al ca1on1aatJon acaampmId an ~bId Pt1ntJ ~
4 JItore larger alcrofauJing (Oiat_ and PzOtoaoana) Gr~_
occurred on the protcbld~
I l
I
bull bull ~
mfEflpoundaS middot~-t9i _jgt- ~ _ _ ~sev M~J~ (1981) (cilonJzatiooof alUf~lng lnts by _rine
Jbull ~teda9g~1ta IIarina XXlV~ 185-191
- - gt 0 Ie 1 _ - _
poundV8I$i I~lf (1~P~h 1II8rin~Ugae and Foul lng AReviw ith
t p~l-i9Jrlr ~iiMdilto5hiP-fQUllfCiH 8ot~ ~r~ ~~~ XXIVbull lIPwl~J11A
Fotteth CN~R ~kk8nGB IWPIR1seJCiiiroaic1lihd ~e ~JIIth CII Offshore St1ucturu ElllS ~tjI~f~1Dt19i4
- - ~
UKC (1976) I~races ill ~W Ecology KIuverd Univbullbull middotraabullf~iddge ~bull C~~jII illt)Ia(t RC (1980) totinllsa Cattod1CPrOtect1cinbull middotmiddot8AtEmiddot~6haOCIlfif~ T~ p No 108 ~ 18 ~ 1IIHf bull
t bull lt ~J~~~ __ ~
Norkr4irulB(198D) Advanculn UcrOfj~lpound~ltiOyVol 4 (AlItxlaquoIder fl Ed) PlerIw Press lcxIdon~ lIP~ 51-85 ~ l~l - bull bull
PJijllt~i At(fm)~mr~~JnI~1fqul1nQ JlalntlJ Riumh ~JnOrpnic toatJngs 2 192middot
)~J gt0 c ~lt -- ~ __ RogerS~ Ut (1968) ina ~(lnt~fbItliP4ili GI COnoslon ~a Technology)bull bull
Sl~iorr~ VP~Ranald~ Rctd~~ (1S80)_ ~ Experllattal Stud1aiRelate middotEffect or Cathod1c Protict1Oi withmiddot terta1nr1c
Coating Syst~ 12th Annualofr T8Id1na1ogy Co1fItHnC8 liauston TIIlt p 9-12
j
fIG1 sPct FromMon IT8TJCuZOI Int Lliu EOAX
FlG~ Fro TlTCu20Ar-4 Dy Field Ellposult I~ir
il
I c
R hi -00amp=7
-I I I I I I I I I f
iii
bull bull
bullbullbull
I J
J
I Figur 3 Thl chan In reltl rqpoIQnlof -ariOtimiddotjOmponents of the paint film with time Thl vtic~1 ampkiS unl till _n ofmiddotmt Illment per unit OJ fe 50all Cl Na
Cu StI Feeshy
PlistinllFe S7~J6 704 3~74 908
f
r e ~ ee~
middotie ~lt - ~ shybullt bull ~
t -~ l Cu~ o ~ f
t --- 3igt - bull 0 J Sn
cc
t ~
~
bullo
~ bull e
bullbullbullbull bull ~
bull ebull a QI 1- bull 0 40 431282 316
101 dmllIII)bullt bullbullbullt
hr 0 ~~bulla 5~16 ~~ -~~ i166 4S i83 8~1B
24tu 6~480 SD~83
111middot 1~1 OS6
~ 5114 6~69 -51 UIamp middot016- lt ~rmiddot
1d o Mea 818 21 021
~Z21 199 1~1Qmiddot ~Jgt2 058
14 d o ~~ 804 Q~B8~~~a~~t~middot 685
866 372amp 8amp4 OS _i~11[ ~~64
Tttile 1 A81UvlL pzQJJortlcw 0 vxl~oI ~~bull Jn 1I1Dtitctel lt-1166) end lqIlDteCt6 (O)middot~af~_~middot_lodEDf~ middotmiddotmiddotmiddotmiddotmiddotmiddotmiddotmiddotc
~ ~
_----I~
J
I bull
I
J bull tmiddot
7o CONQRESOIMTERNACIONAL
I D~CORRO$IOr(IAAINAEmiddot~USTACIONES~v c)
t 7thflfERHATlONAL CONGRESS
ON IARINE CORROSION AND fO(JUNG
~CONGRE$INtERNATIONAL DE~CORR~ON ~E ET QES~SAUSSURE$
i t
~ ~
bullt
t t
UNIVERSIDAD PourtcNlCAVollncio7J1 NiwiemIif~ 1988
(SPAN
SECcl6lfn
BiOIogro marino Morine biologY ~1i1orine
Theunpub1ishedJllBIlu8criptEl have been reprod~d asr~ielved from the
authors The SCientiUc COlIIIBittee take not respoi~ulbll~~Y for any shy
error or omssi9n
Laa ponenciasno han side publicadas y estanreproducidas tal y como
sellan recibido demiddot BUSmiddot autorea pound1 Conte Cientfico middotno se responsemiddot
bil1za de cualquier error ~ om1si~n
bullbull _ bullbull_ bullbull ___ bull $_ _ L _ - _ --- -__
SESSION II Marine biology
Friday 11th November
SURFACE PROPERTIES ON NONmiddotTOXIC ANTIFOULING PAINT FILM
K Nanishi M Murase Y Yonehara M Kishihara and T Hirama
MICROFOULING ON ALUMINIUM PANELS PLACED IN THE TROPICAL WATERS
N B Bhosle K Venkat and A B Wagh
EFFECT OF TRIBUTYLTIN OXIDE (TBTO) ON THE OVARIAN MATURATION OF THE PRAWN CARIDINA WEBERI
R Nagabhushanam B Indira R Sarojini and Mary Frances Thompson
EFFECT OF BIOACTIVE MATERIALS FROM SOFT CORALS ON BACTERIA ASSOCIATED WITH COMMON FOULER
Vitalina Mary Avelin Mary R Sarojini and R Nagabhushanam
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
bull bull ~
~ ~ - f
bull ~ _T~~-middot~ ~~- ~r _ -$1
(1) ~rlttionor Painted Steel Billets middot_ ~~tmiddot_tlJ middot~hmiddot~--~fmiddot ~ middot~A~-~~middot - -~ ~
Thirty-al ateel blllets (8-10aD2 end l depth) were cut frollbullt ~middot~middot~middotmiddot~middottrSmiddott ~jS-~ It bull gt~ tfl - middot--imiddotmiddot ~ t1dItrd steal plates The dimenSlonsof -thB butits-lim chosento
bull -~~ middotmiddotgt2h ~ ~-Imiddot ~~rt ~- -middoti~~ ni ~~middot1 l middot-~t~~middot _
(a) fecilltate amiddot Ie detrucUve -thxI of prePraUonpdor to ~~_gt gt~ k7i ~_ t~ 1--1- _ ) - +--_~ -_~bull ming electron ~ 4II1CI (iialloWdirectobs8iVatlCnS of the
- i lt-- ~ gtbull -o1e billet 8Uface
gt r~middotmiddotj ~L(~middot~middot rmiddot tmiddot J ~ - ~ - lt _ bull Single non-standard (1hIula~) nickel lIIireSmiddot Spot Welded e1Cing
bull ~1deJ ~ b~i~ ~j~~ tri afl~1b1ipOi~ f ~1t1 ~4middotmiddot f-gt -~ ~S-- ( -- - - _bull resin EactI billet then Undb~ tI1d prtiladlllltha5O __ eyilr
bull ~~_ ~~imiddot ~1iil h~ intlfcili~ ~eonbt1nlyi Tr1butylt1n F~id(18n)~~~O~dettu20i~blcicld8is 1IIfii bull i - ~ ( then 1IPI)l1ed to glvaadry Ulil tIi1c1ltnOss or 1(1)-150bullbull
tbull (2) IipreSSeCi Current Syat
bull A rigid ~tic due 1I8S drWedllliihwrtv-s41 liiIihOieS IIPProX~
bullbull 05-10 cia episrt ~ Pilnted ampUl8t Wsfbed IntaPQSIUon ampy
threadlng the nickel IIIlr8throu11 the dinto a tarproot junction
box Eliltilen of th8~1l1ii (recelving ~t-d curnritlere IIIlred
Into bull block connectOrrecelvlng dc trOll is potentiOstat The
rainJng iontroi blUets Wet8fbedfl11i1iiy lIIithln thejtlytlonbox
i~ityd~c IkiliPly ~~ ~lat1ruaWiode 8ncIacalOliel
refer~ electtodetoiere fllied ~ the per1aetat of the plut1cdlc IIIId
connected to the pot8ritlostat The desl~ of the potentlosbt ailoiolEld
~PlDteItlO1TClJrnnt to be MIllntained at 8prampset levtl of 966 II1II
c
r~
Grid raf 1J5s-t 114iH 563724 dud 0 Juna
Alolecule andbechr1a in general are ~ to
sutface lI1eroleyer (nlUSton) of _ter (~bullbull 191KJ)~To avold
contaJnatlonof ttw painted surf by U- -vents
blllets Were s-nact in a 1 conte1ner or atc1le (02i ) f~~nd
ter Iih1ch then f9ICI in tv 1IIhere the b~
be r-oved undeNeter El~~ bllleta fa Mcb trtai bullbullel
removed over a 1 dtq (336 hr~ expoeure perioda
each being ~~bn uposurnof 1 5 11 iA 168and DlIIauz1r
Arter field axpauHli the billets wan
fUtered _tar to ~ 1_ debtlflMd far 10 aJnuta ~
wv ~~~endattid sdbUiUd(otO
~ ~l f~ nn1ng el~~ --
lOOted onto 1middotCIi ~bullbullWIsend carbonmiddotcoa~nmiddotmiddot~middot f
each billet Ws~ Iis1ng a ccr1dge ~el~~
(5360) IIIlth anergy dJ8perslve X-ray alcroenelre1a (mx)
_lysis the les Wen reNed end coated III1th vold to al]Ql En
detailed sat of the alaquofa1l
(4) Lfiborattux ~
Painted taa~ bUletalllllre pnparad ~
in a Mrlne reclrculet1ng flOlll _ ~
f1Oll the ~ ~a1t~y Tha~~Un bullybullbullbull
10C M1nta1ned ~~t1as of ~t~la protazOllbull~bull10shy
(Entermap) ~ ~~~middotNlt~Eamp1~ __~and ~ r - ~ -
certaJn ~lc 1IQ1~ 1(1 the
the painted
~
thre repl1catft fn
plKed 1nto(022_)
- thIine1J~~ -lt bull
Atter mAX
(1-2) and i-n-t bn collected
14OC
Pain~
-
12 daY~ exposure and prepared as in d)~ - t
~I ~~~-- i~yen 1~ v
Rflltsi ~ _ ~f-~lI ft j bull _
rt~ll0~~~llY~~~~peln~stetsl~11i~ex~sed for
y~t1~J~ ~~~~1~1~~1UII~ya~kooJ~~~lt ~ ~~_~~I1J~ ~~~tomAX -lysis ~era than gold ~ bull bullbullbull c - _ bull lt
~~fllr ~~~ vllUll ~t1ans of the rfee htures and gt Co
~f~ - ~~~ ~f ~ ~aobta1Mdue IjIll811 iI1Flg 1~2 The IIIIdn ~~ ~ lt)ji9 )tmiddotmiddot - ht
- bull
~I rIIplt~ tIt8 -~ cOlllponanta of the pei1t 1bullbull WelIIlItr1x $ilj~t-~~middot - ~ _ -_
-IUch PVCbQed end rap~13y the chlorlna peak the
i~~~-~~-~V~~~-t18 ~~~~ ~~ Th8
ral~~i~ JlrQP(lrUo1sof ch 1Il-ntal paek~ll1d l~t_ Q~7 -gt- ~ _~ -- gt ) gt -_ -lt ~~ the 0 -end _ xprtsntabullTIwsa changes are -r1sed ~t~ lt~ _ ~ 1nT~ IThe8llia$lb npresentthe chengelI 1n~t1V8 p~lons
ti~~~ [ ~gt 3 - - - _ - _ -
Qf nellllllllt within We surface layars of the pe1ntmiddot ru The
~f~~thtJn -d ~rfeli avr the 336 h11cxI ampScanbe
SII4iIl r~ fig 3 al~ tht rta of~ dirfers tJetwaeo thaa ltmiddot~1- 1ft~- ~ _
IJOl appfiai-ed to ~1nlt1elly until ftar day 168 it fall
~~l~ 1h8relllllrelgn1fi~ diff~be~ qrOtactlid and
proieCted (at -966 ) piIIllnts tor lioo COPPer but not folimiddottin
Ibi~tlons Of the Pain -urface 8hoWad _Pil ~ witilt1M
(Plete I to IV)~ 1hePllat1na (~) pelnt(Phta x and b)
Clearl~st-i~~r alots on the SurfllC8 IiII1kh~ter1sed ~- f hlt1 lriitUl tin conCantrat1on~ lllariy slotsre Partially
~OIIe~ ~ tile _tr~tedal middot1lere rs ~an~ densely packed
grerltr~ ~th 1ar~(iIa1niy cotifIIr) 8ntt -ampniBil (lIB1rily 1rori)being
middot vl~l~~~~~ Dtt~i~~~t~ r~ ~f~~ I~t seawatllr for 11 n IIIOre slots appeared and th~ ~~ess _trb cwer
--if -- V~ c~~ f(i ~~~i~
(Plate Ie to It)~ the granules dld not protrude from the~rtace in ~ ~~~~j1~r~-ffJmiddot0~~
suhap~~r Both 0 nd --960 at ~~pa1nts ~ - fr ~~~~-w1-U~ ~~t
sl~llrat thi stag~ (Plates Ic-d II1d xrnspac~IlYh middotmiddotWith middot middotaJ~cmiddot gt incraallO ~alolre t~ difrrqcbullbull middotbatIiI8IImiddot the protected fjd
bull ~ ~ ~t~
IIllrotacbldpaint ~~lIfIt~ftitr~ h alQt~ p~t bull u bullbullbull J ~~i~~ 1~~
lIIith IIIuper lIdguon u 4lrotatad piIJnt (Pl8ti1 II) 1hD1II8S
Ill )1IMdw1lillS gnnJla~on f-r~~ dpert1cl ~ - - - ~~r ~t
Onthe 966 1IIpetnt t-var tIIO typeaofan with diffannt c t
~hIId~(Plate UI a to c) One hIId tIII-1 bull bullbull e _ bull
fUll wJ1hleas abvlaus alats the o1fwl bed C18UU1IlOt ~litUi middotC c ~ ~
less ~ t11 h Gnnulatlan __ but ~lIiIZly def1nlld in
middot~ latter Afte fila bed tiIIrtIId 10 ~are ~~j(
extansll(tlly ~ the~tIId(O at) tIIIIple lIIIdngthe~ - ~
middot orarallet1ai leas claaI (Plate I~) f u II1ao CQ1tJAJ11d to ~Gn - -
the -966 at 1 (Plata III d tor but 1JI8V8n 1nittNdcneabull -
Althatqa -middotslots wrs abvlaus becaI1ng 1 cwampr aiI- the orwWat1an8
f1nailyftar 14 day exposure ~ fUll drMIlapIIIent on all la - ~
are extensive with the 0 lhawlng both alota pore ~
OVIKgrOloln (Plata II c to f) Gnrulatlon 1bl to ~ gt~t~~ i
or the th1cknats of tho ma ~the ~ at ~ (~~~~ themiddot - - _ middott ~ ~ ~ - -1 1 t
middot f11bull IIIIIS IIOre extans1 thIIIn at dIlya but IIIIIIIY bull1Q~ tI st11l ~w -middotr~ 1gt ~1 ~
obvious fIIany ~ loI8re still p~ yen~~~~~~~r
than before Poiu appearedto be f~lf~ila~~c-~~~a (Plate IVf~ label 1) IIIIl1ch broke~i(2)~ ~~~~
t t
bull ~lS (3) ~ rflC~~ ~i~t~~l~~KI~ t~ldiIiiicyul~loIiih ~~Rht ~ )~- -~ - ~middotd 40 ( gt ~gt ~d lt1 t ~ Jt ~remiddot4amp tha paint trix cavity bottom eroded away reVe81~ng ore
t ~~t~ por~~ ~l~ P~]Y p~~8dby di~lutiaA~ ~~~~ti~Jr~gt~ta(l~~Siinll1cl1Ud~f iUiIiI reriatlGllbullt
bull a~~ ba~tid colCriAtloriccic~ GIl ~~~d ri -966 - ib~~~amp catQn1iti~~t~I~~(~t an ( ~
t the ~~~palnts (PIa_taiiI d~~h ~wh ~ n td have ~ CamplluJMitiIVc) ~~middottti o~lgtnalrf tlt of hIght
~ l=~A ~J~rW tYtles~ p~fiiilriQcOCCold to~Ja to jU-t ~jmiddoti~ai dlatClilS ~ir -jlati ~tbullbullent aNit ~ t ~~ th8middotf~ld~ies~~t~on~ ~ -966 prQ~~~(P14ta Iii~)Omiddotiliof~m1hiiHAiiipieat ere~ll-(piitir~ p-iot~iilans(PiA-iII~ end c) nbullbulltiledt dl~by1i~~ end wirinali1sheii ~cMI~bY thI tili
bull In t-~~ ~1i1nt 1oIIIeHfd~ ~m1IS (15OClind a t ri~r-dlatoa ~lor P~t dlataiOlonlt1anGIl ttw-966
t
bull ~
pr~t~ ~is~ih iion obitJauIJ A ~rot -=1 ~ ~ i __
reprasenttild on the 966 painted 1iUrfc1ncluicunljj ~ rwt
Achnanths Colonisation by bacteria 1n the laboretory bsed
bull eliperlMnt tollMct sbiUar pattern to the fild ~
bull bull Dilicueii
bull bull DUt 1Iidnt ~lona Mve dlttllrint ~~ ictian
It ~ JntM l1tenture ttt Iff yen __ JIll the
bull ~ bicins ani 111 ~t INchlnil (PhillIPs 1t73t EWene~ 1981
In ci1ntlHt tJWY ~tit ~Wl1c iIuCh as 1rbiltyl tirI
~ lite wAlII]ltiClr dlsP8tfu~ middotthiirsttilll~intaJn~ a
bull ~
lIIifoflll ampalCentraUon through the filII 10 thoIt biocide rel_ at tha
paint surfce 1fI$Jlts in IIIOre ditfusing ~s to repla~lj -iflJ~it
resulting 111 bullbulltNdy lr leechlng rata M1llat this IIIIY be middottruIi ot
tin ~ it was nOt true ~ ttw middotlBlf fOZWJlatiGn ua ~ bull - i - bull 1(1
ObSfvt1onundr SE clarly ~ tM t1l1 to be present
~ly lIS OIIIdle-llk parUc1~~t tn lUlfiice (Ito
IV) bUt usa ~ the dIpth attt fU-middot ~y~
If one considra the concn~rtlonmiddot chMge 111 1n ~111c COKICIl8rlts fro- the paint tU- i bullbullbull tMWei b~ ~ Si6t arid
tI-~t iron oxidt 1~ can bt (Fl_ 3 IrId TIibl8I) ~t ~ ase cona1derllble d1fftrenC8s ~ a Un decz bcdYMr ttw p1iod IIIIwraS lron~- an unt Ught inCr h ~ incqJper ~tlGn occUrnd _idly at ttw ~~ Gfb ~ period ~ out ttwn fu nPldly ampQII1n ~ umiddotirid II phast This IIIOUld bt tIlq)8Ctad It cantact l-=h1ng we occun1ng
IIltfIOS8d pertlcl of copper would lMch rapldly on t-n1on 1n - shy JIsewtr There would thlll1 ba bull J-a a1iltUy leu avalUba
i1 ~ ~
_tedal becfM uposed Thi tIOUld ttwn IIllCh Thla pulSing efttct
IIIOUld be expctad to -oth OIItlllith U- putic1 __ -lllible II tot 1~ GIl a on r-x _lao Thla teet 01 put1c1e 1tUng
can tit GIl the lIUrlace ClIlC8 it ~ a pan 1amp t
a 11aM bet in Plat I to IV IIaIrIg _ lit la_ II ~ 0- ~s _- PIate IW $llldl ~bull nshyot poRI flllNltttn Al~ tin ~ lou rete thtn I bullbullbull atill Ivwnce of bull al~t pulsed 1Itue rrlict tII1IIWd
elqlCtaG inIl ttwauUr ~ I I I
0
~ i
~ ~
~I J
bullbullt bullbullbullbullt bull bull ~
bull bull
I
bull t
~~
The same seqiHllle of cevity follliltloii (intttecaseofthetestjjltle
slot- shaped) fClliOileciby SecOlld8rYtlolesfotlllingin thll cavity
bottollis releasing further deep8rtox1ils (Plate XV d and g) As
cavities_re ProdICed ~ tiecaaIelnvadedby_ter~ onelilauld
expect 1ncreaa 1n1IOdha and dIladne lMls t the t1met
which the toxlM uw ~ Ths fCMld tCtHi eo (Flvmiddot 3)
~rttno the vi that_tat ~t1on otthe~ ~lY
il1Cl1l8S8S Idth UIiJ
Differences f~in toxin li~~to Ini 966 ill 8811P~ F ~ bOth Un end ~ lcMIr levels afillW
IIIIIlfI ptaInt(~1~~1ntha~~ ~~f~~ cst rklld with tbiIccpper ElJIc~l ta~Uona invalvinv
w pl8cId at a~ht~~middot3lO~lth Un cqJpel bull
(-960)10 ttW ~af this expei1tIIri1ts thIsiorelfJltdt -
that cqlp8r b8IiIId ~ IIiIOUldProc-ln f41ip1dly t thellllgllitiv
patentW _lied () as 1t 11 further f~1t ~ witt Un
III11ch is u-tat aqulllbd Idth
it If ttda 18 eo~ ~
~ )
frOll1 paintscould bl anqnced by an illprd current clithodic
protecUansysteipos1bly ahart_nino te 11fe of the paInts
antifouling ability Bacterial coloni Uon IIIU rtdUCed on the -s66 paint This
could have been due to unflSllOUlllble electrostatic cqrge on th surtece Th1a effect CIIl pley en IIIportant role in the t~of
bacteria to alrraces (lIIarsMll19E1O) As bacteria appear to IhiJw
greater toxicltyto copper (DMpsey 1981) it is Klre likely that
enhanced cqJper leacll1nl On the -966 AIIIPl c8used the recluCed
activity The IDlol8r toxicity ofUn to bacteria (as reported by
~ -
DelIIPsey Hle1gt or its lowes leaching rat was sUpPorted bythl
relatively rapid recolonisation of the slots by lIIicroorganiSlllS ~ftar thE inlt1al aurge of leachi~ Thailllpressecl current dto ~~ the oppoSite _ff~middot ~r on dtta colonstJon l1lI-1- in the
JIbcJrtory~ta larger rUDIIra f~ fit pro~td ~ther than uriprotected Pafnta
Ccnlus1one
1 tiild-a 18 presaIIUd ~ thattJn~ ~~
~llNChng notd1rf~ ~
2 TIleuseofCP~ ~CUIftnt(at) on1h1a ~fca41ng ~- ~
pa1I]t~ to ~_lAIIach1ngn_ at toxiN - -
3 leIJstaPidbectar1al ca1on1aatJon acaampmId an ~bId Pt1ntJ ~
4 JItore larger alcrofauJing (Oiat_ and PzOtoaoana) Gr~_
occurred on the protcbld~
I l
I
bull bull ~
mfEflpoundaS middot~-t9i _jgt- ~ _ _ ~sev M~J~ (1981) (cilonJzatiooof alUf~lng lnts by _rine
Jbull ~teda9g~1ta IIarina XXlV~ 185-191
- - gt 0 Ie 1 _ - _
poundV8I$i I~lf (1~P~h 1II8rin~Ugae and Foul lng AReviw ith
t p~l-i9Jrlr ~iiMdilto5hiP-fQUllfCiH 8ot~ ~r~ ~~~ XXIVbull lIPwl~J11A
Fotteth CN~R ~kk8nGB IWPIR1seJCiiiroaic1lihd ~e ~JIIth CII Offshore St1ucturu ElllS ~tjI~f~1Dt19i4
- - ~
UKC (1976) I~races ill ~W Ecology KIuverd Univbullbull middotraabullf~iddge ~bull C~~jII illt)Ia(t RC (1980) totinllsa Cattod1CPrOtect1cinbull middotmiddot8AtEmiddot~6haOCIlfif~ T~ p No 108 ~ 18 ~ 1IIHf bull
t bull lt ~J~~~ __ ~
Norkr4irulB(198D) Advanculn UcrOfj~lpound~ltiOyVol 4 (AlItxlaquoIder fl Ed) PlerIw Press lcxIdon~ lIP~ 51-85 ~ l~l - bull bull
PJijllt~i At(fm)~mr~~JnI~1fqul1nQ JlalntlJ Riumh ~JnOrpnic toatJngs 2 192middot
)~J gt0 c ~lt -- ~ __ RogerS~ Ut (1968) ina ~(lnt~fbItliP4ili GI COnoslon ~a Technology)bull bull
Sl~iorr~ VP~Ranald~ Rctd~~ (1S80)_ ~ Experllattal Stud1aiRelate middotEffect or Cathod1c Protict1Oi withmiddot terta1nr1c
Coating Syst~ 12th Annualofr T8Id1na1ogy Co1fItHnC8 liauston TIIlt p 9-12
j
fIG1 sPct FromMon IT8TJCuZOI Int Lliu EOAX
FlG~ Fro TlTCu20Ar-4 Dy Field Ellposult I~ir
il
I c
R hi -00amp=7
-I I I I I I I I I f
iii
bull bull
bullbullbull
I J
J
I Figur 3 Thl chan In reltl rqpoIQnlof -ariOtimiddotjOmponents of the paint film with time Thl vtic~1 ampkiS unl till _n ofmiddotmt Illment per unit OJ fe 50all Cl Na
Cu StI Feeshy
PlistinllFe S7~J6 704 3~74 908
f
r e ~ ee~
middotie ~lt - ~ shybullt bull ~
t -~ l Cu~ o ~ f
t --- 3igt - bull 0 J Sn
cc
t ~
~
bullo
~ bull e
bullbullbullbull bull ~
bull ebull a QI 1- bull 0 40 431282 316
101 dmllIII)bullt bullbullbullt
hr 0 ~~bulla 5~16 ~~ -~~ i166 4S i83 8~1B
24tu 6~480 SD~83
111middot 1~1 OS6
~ 5114 6~69 -51 UIamp middot016- lt ~rmiddot
1d o Mea 818 21 021
~Z21 199 1~1Qmiddot ~Jgt2 058
14 d o ~~ 804 Q~B8~~~a~~t~middot 685
866 372amp 8amp4 OS _i~11[ ~~64
Tttile 1 A81UvlL pzQJJortlcw 0 vxl~oI ~~bull Jn 1I1Dtitctel lt-1166) end lqIlDteCt6 (O)middot~af~_~middot_lodEDf~ middotmiddotmiddotmiddotmiddotmiddotmiddotmiddotmiddotc
~ ~
_----I~
J
I bull
I
J bull tmiddot
7o CONQRESOIMTERNACIONAL
I D~CORRO$IOr(IAAINAEmiddot~USTACIONES~v c)
t 7thflfERHATlONAL CONGRESS
ON IARINE CORROSION AND fO(JUNG
~CONGRE$INtERNATIONAL DE~CORR~ON ~E ET QES~SAUSSURE$
i t
~ ~
bullt
t t
UNIVERSIDAD PourtcNlCAVollncio7J1 NiwiemIif~ 1988
(SPAN
SECcl6lfn
BiOIogro marino Morine biologY ~1i1orine
Theunpub1ishedJllBIlu8criptEl have been reprod~d asr~ielved from the
authors The SCientiUc COlIIIBittee take not respoi~ulbll~~Y for any shy
error or omssi9n
Laa ponenciasno han side publicadas y estanreproducidas tal y como
sellan recibido demiddot BUSmiddot autorea pound1 Conte Cientfico middotno se responsemiddot
bil1za de cualquier error ~ om1si~n
bullbull _ bullbull_ bullbull ___ bull $_ _ L _ - _ --- -__
SESSION II Marine biology
Friday 11th November
SURFACE PROPERTIES ON NONmiddotTOXIC ANTIFOULING PAINT FILM
K Nanishi M Murase Y Yonehara M Kishihara and T Hirama
MICROFOULING ON ALUMINIUM PANELS PLACED IN THE TROPICAL WATERS
N B Bhosle K Venkat and A B Wagh
EFFECT OF TRIBUTYLTIN OXIDE (TBTO) ON THE OVARIAN MATURATION OF THE PRAWN CARIDINA WEBERI
R Nagabhushanam B Indira R Sarojini and Mary Frances Thompson
EFFECT OF BIOACTIVE MATERIALS FROM SOFT CORALS ON BACTERIA ASSOCIATED WITH COMMON FOULER
Vitalina Mary Avelin Mary R Sarojini and R Nagabhushanam
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
-
12 daY~ exposure and prepared as in d)~ - t
~I ~~~-- i~yen 1~ v
Rflltsi ~ _ ~f-~lI ft j bull _
rt~ll0~~~llY~~~~peln~stetsl~11i~ex~sed for
y~t1~J~ ~~~~1~1~~1UII~ya~kooJ~~~lt ~ ~~_~~I1J~ ~~~tomAX -lysis ~era than gold ~ bull bullbullbull c - _ bull lt
~~fllr ~~~ vllUll ~t1ans of the rfee htures and gt Co
~f~ - ~~~ ~f ~ ~aobta1Mdue IjIll811 iI1Flg 1~2 The IIIIdn ~~ ~ lt)ji9 )tmiddotmiddot - ht
- bull
~I rIIplt~ tIt8 -~ cOlllponanta of the pei1t 1bullbull WelIIlItr1x $ilj~t-~~middot - ~ _ -_
-IUch PVCbQed end rap~13y the chlorlna peak the
i~~~-~~-~V~~~-t18 ~~~~ ~~ Th8
ral~~i~ JlrQP(lrUo1sof ch 1Il-ntal paek~ll1d l~t_ Q~7 -gt- ~ _~ -- gt ) gt -_ -lt ~~ the 0 -end _ xprtsntabullTIwsa changes are -r1sed ~t~ lt~ _ ~ 1nT~ IThe8llia$lb npresentthe chengelI 1n~t1V8 p~lons
ti~~~ [ ~gt 3 - - - _ - _ -
Qf nellllllllt within We surface layars of the pe1ntmiddot ru The
~f~~thtJn -d ~rfeli avr the 336 h11cxI ampScanbe
SII4iIl r~ fig 3 al~ tht rta of~ dirfers tJetwaeo thaa ltmiddot~1- 1ft~- ~ _
IJOl appfiai-ed to ~1nlt1elly until ftar day 168 it fall
~~l~ 1h8relllllrelgn1fi~ diff~be~ qrOtactlid and
proieCted (at -966 ) piIIllnts tor lioo COPPer but not folimiddottin
Ibi~tlons Of the Pain -urface 8hoWad _Pil ~ witilt1M
(Plete I to IV)~ 1hePllat1na (~) pelnt(Phta x and b)
Clearl~st-i~~r alots on the SurfllC8 IiII1kh~ter1sed ~- f hlt1 lriitUl tin conCantrat1on~ lllariy slotsre Partially
~OIIe~ ~ tile _tr~tedal middot1lere rs ~an~ densely packed
grerltr~ ~th 1ar~(iIa1niy cotifIIr) 8ntt -ampniBil (lIB1rily 1rori)being
middot vl~l~~~~~ Dtt~i~~~t~ r~ ~f~~ I~t seawatllr for 11 n IIIOre slots appeared and th~ ~~ess _trb cwer
--if -- V~ c~~ f(i ~~~i~
(Plate Ie to It)~ the granules dld not protrude from the~rtace in ~ ~~~~j1~r~-ffJmiddot0~~
suhap~~r Both 0 nd --960 at ~~pa1nts ~ - fr ~~~~-w1-U~ ~~t
sl~llrat thi stag~ (Plates Ic-d II1d xrnspac~IlYh middotmiddotWith middot middotaJ~cmiddot gt incraallO ~alolre t~ difrrqcbullbull middotbatIiI8IImiddot the protected fjd
bull ~ ~ ~t~
IIllrotacbldpaint ~~lIfIt~ftitr~ h alQt~ p~t bull u bullbullbull J ~~i~~ 1~~
lIIith IIIuper lIdguon u 4lrotatad piIJnt (Pl8ti1 II) 1hD1II8S
Ill )1IMdw1lillS gnnJla~on f-r~~ dpert1cl ~ - - - ~~r ~t
Onthe 966 1IIpetnt t-var tIIO typeaofan with diffannt c t
~hIId~(Plate UI a to c) One hIId tIII-1 bull bullbull e _ bull
fUll wJ1hleas abvlaus alats the o1fwl bed C18UU1IlOt ~litUi middotC c ~ ~
less ~ t11 h Gnnulatlan __ but ~lIiIZly def1nlld in
middot~ latter Afte fila bed tiIIrtIId 10 ~are ~~j(
extansll(tlly ~ the~tIId(O at) tIIIIple lIIIdngthe~ - ~
middot orarallet1ai leas claaI (Plate I~) f u II1ao CQ1tJAJ11d to ~Gn - -
the -966 at 1 (Plata III d tor but 1JI8V8n 1nittNdcneabull -
Althatqa -middotslots wrs abvlaus becaI1ng 1 cwampr aiI- the orwWat1an8
f1nailyftar 14 day exposure ~ fUll drMIlapIIIent on all la - ~
are extensive with the 0 lhawlng both alota pore ~
OVIKgrOloln (Plata II c to f) Gnrulatlon 1bl to ~ gt~t~~ i
or the th1cknats of tho ma ~the ~ at ~ (~~~~ themiddot - - _ middott ~ ~ ~ - -1 1 t
middot f11bull IIIIIS IIOre extans1 thIIIn at dIlya but IIIIIIIY bull1Q~ tI st11l ~w -middotr~ 1gt ~1 ~
obvious fIIany ~ loI8re still p~ yen~~~~~~~r
than before Poiu appearedto be f~lf~ila~~c-~~~a (Plate IVf~ label 1) IIIIl1ch broke~i(2)~ ~~~~
t t
bull ~lS (3) ~ rflC~~ ~i~t~~l~~KI~ t~ldiIiiicyul~loIiih ~~Rht ~ )~- -~ - ~middotd 40 ( gt ~gt ~d lt1 t ~ Jt ~remiddot4amp tha paint trix cavity bottom eroded away reVe81~ng ore
t ~~t~ por~~ ~l~ P~]Y p~~8dby di~lutiaA~ ~~~~ti~Jr~gt~ta(l~~Siinll1cl1Ud~f iUiIiI reriatlGllbullt
bull a~~ ba~tid colCriAtloriccic~ GIl ~~~d ri -966 - ib~~~amp catQn1iti~~t~I~~(~t an ( ~
t the ~~~palnts (PIa_taiiI d~~h ~wh ~ n td have ~ CamplluJMitiIVc) ~~middottti o~lgtnalrf tlt of hIght
~ l=~A ~J~rW tYtles~ p~fiiilriQcOCCold to~Ja to jU-t ~jmiddoti~ai dlatClilS ~ir -jlati ~tbullbullent aNit ~ t ~~ th8middotf~ld~ies~~t~on~ ~ -966 prQ~~~(P14ta Iii~)Omiddotiliof~m1hiiHAiiipieat ere~ll-(piitir~ p-iot~iilans(PiA-iII~ end c) nbullbulltiledt dl~by1i~~ end wirinali1sheii ~cMI~bY thI tili
bull In t-~~ ~1i1nt 1oIIIeHfd~ ~m1IS (15OClind a t ri~r-dlatoa ~lor P~t dlataiOlonlt1anGIl ttw-966
t
bull ~
pr~t~ ~is~ih iion obitJauIJ A ~rot -=1 ~ ~ i __
reprasenttild on the 966 painted 1iUrfc1ncluicunljj ~ rwt
Achnanths Colonisation by bacteria 1n the laboretory bsed
bull eliperlMnt tollMct sbiUar pattern to the fild ~
bull bull Dilicueii
bull bull DUt 1Iidnt ~lona Mve dlttllrint ~~ ictian
It ~ JntM l1tenture ttt Iff yen __ JIll the
bull ~ bicins ani 111 ~t INchlnil (PhillIPs 1t73t EWene~ 1981
In ci1ntlHt tJWY ~tit ~Wl1c iIuCh as 1rbiltyl tirI
~ lite wAlII]ltiClr dlsP8tfu~ middotthiirsttilll~intaJn~ a
bull ~
lIIifoflll ampalCentraUon through the filII 10 thoIt biocide rel_ at tha
paint surfce 1fI$Jlts in IIIOre ditfusing ~s to repla~lj -iflJ~it
resulting 111 bullbulltNdy lr leechlng rata M1llat this IIIIY be middottruIi ot
tin ~ it was nOt true ~ ttw middotlBlf fOZWJlatiGn ua ~ bull - i - bull 1(1
ObSfvt1onundr SE clarly ~ tM t1l1 to be present
~ly lIS OIIIdle-llk parUc1~~t tn lUlfiice (Ito
IV) bUt usa ~ the dIpth attt fU-middot ~y~
If one considra the concn~rtlonmiddot chMge 111 1n ~111c COKICIl8rlts fro- the paint tU- i bullbullbull tMWei b~ ~ Si6t arid
tI-~t iron oxidt 1~ can bt (Fl_ 3 IrId TIibl8I) ~t ~ ase cona1derllble d1fftrenC8s ~ a Un decz bcdYMr ttw p1iod IIIIwraS lron~- an unt Ught inCr h ~ incqJper ~tlGn occUrnd _idly at ttw ~~ Gfb ~ period ~ out ttwn fu nPldly ampQII1n ~ umiddotirid II phast This IIIOUld bt tIlq)8Ctad It cantact l-=h1ng we occun1ng
IIltfIOS8d pertlcl of copper would lMch rapldly on t-n1on 1n - shy JIsewtr There would thlll1 ba bull J-a a1iltUy leu avalUba
i1 ~ ~
_tedal becfM uposed Thi tIOUld ttwn IIllCh Thla pulSing efttct
IIIOUld be expctad to -oth OIItlllith U- putic1 __ -lllible II tot 1~ GIl a on r-x _lao Thla teet 01 put1c1e 1tUng
can tit GIl the lIUrlace ClIlC8 it ~ a pan 1amp t
a 11aM bet in Plat I to IV IIaIrIg _ lit la_ II ~ 0- ~s _- PIate IW $llldl ~bull nshyot poRI flllNltttn Al~ tin ~ lou rete thtn I bullbullbull atill Ivwnce of bull al~t pulsed 1Itue rrlict tII1IIWd
elqlCtaG inIl ttwauUr ~ I I I
0
~ i
~ ~
~I J
bullbullt bullbullbullbullt bull bull ~
bull bull
I
bull t
~~
The same seqiHllle of cevity follliltloii (intttecaseofthetestjjltle
slot- shaped) fClliOileciby SecOlld8rYtlolesfotlllingin thll cavity
bottollis releasing further deep8rtox1ils (Plate XV d and g) As
cavities_re ProdICed ~ tiecaaIelnvadedby_ter~ onelilauld
expect 1ncreaa 1n1IOdha and dIladne lMls t the t1met
which the toxlM uw ~ Ths fCMld tCtHi eo (Flvmiddot 3)
~rttno the vi that_tat ~t1on otthe~ ~lY
il1Cl1l8S8S Idth UIiJ
Differences f~in toxin li~~to Ini 966 ill 8811P~ F ~ bOth Un end ~ lcMIr levels afillW
IIIIIlfI ptaInt(~1~~1ntha~~ ~~f~~ cst rklld with tbiIccpper ElJIc~l ta~Uona invalvinv
w pl8cId at a~ht~~middot3lO~lth Un cqJpel bull
(-960)10 ttW ~af this expei1tIIri1ts thIsiorelfJltdt -
that cqlp8r b8IiIId ~ IIiIOUldProc-ln f41ip1dly t thellllgllitiv
patentW _lied () as 1t 11 further f~1t ~ witt Un
III11ch is u-tat aqulllbd Idth
it If ttda 18 eo~ ~
~ )
frOll1 paintscould bl anqnced by an illprd current clithodic
protecUansysteipos1bly ahart_nino te 11fe of the paInts
antifouling ability Bacterial coloni Uon IIIU rtdUCed on the -s66 paint This
could have been due to unflSllOUlllble electrostatic cqrge on th surtece Th1a effect CIIl pley en IIIportant role in the t~of
bacteria to alrraces (lIIarsMll19E1O) As bacteria appear to IhiJw
greater toxicltyto copper (DMpsey 1981) it is Klre likely that
enhanced cqJper leacll1nl On the -966 AIIIPl c8used the recluCed
activity The IDlol8r toxicity ofUn to bacteria (as reported by
~ -
DelIIPsey Hle1gt or its lowes leaching rat was sUpPorted bythl
relatively rapid recolonisation of the slots by lIIicroorganiSlllS ~ftar thE inlt1al aurge of leachi~ Thailllpressecl current dto ~~ the oppoSite _ff~middot ~r on dtta colonstJon l1lI-1- in the
JIbcJrtory~ta larger rUDIIra f~ fit pro~td ~ther than uriprotected Pafnta
Ccnlus1one
1 tiild-a 18 presaIIUd ~ thattJn~ ~~
~llNChng notd1rf~ ~
2 TIleuseofCP~ ~CUIftnt(at) on1h1a ~fca41ng ~- ~
pa1I]t~ to ~_lAIIach1ngn_ at toxiN - -
3 leIJstaPidbectar1al ca1on1aatJon acaampmId an ~bId Pt1ntJ ~
4 JItore larger alcrofauJing (Oiat_ and PzOtoaoana) Gr~_
occurred on the protcbld~
I l
I
bull bull ~
mfEflpoundaS middot~-t9i _jgt- ~ _ _ ~sev M~J~ (1981) (cilonJzatiooof alUf~lng lnts by _rine
Jbull ~teda9g~1ta IIarina XXlV~ 185-191
- - gt 0 Ie 1 _ - _
poundV8I$i I~lf (1~P~h 1II8rin~Ugae and Foul lng AReviw ith
t p~l-i9Jrlr ~iiMdilto5hiP-fQUllfCiH 8ot~ ~r~ ~~~ XXIVbull lIPwl~J11A
Fotteth CN~R ~kk8nGB IWPIR1seJCiiiroaic1lihd ~e ~JIIth CII Offshore St1ucturu ElllS ~tjI~f~1Dt19i4
- - ~
UKC (1976) I~races ill ~W Ecology KIuverd Univbullbull middotraabullf~iddge ~bull C~~jII illt)Ia(t RC (1980) totinllsa Cattod1CPrOtect1cinbull middotmiddot8AtEmiddot~6haOCIlfif~ T~ p No 108 ~ 18 ~ 1IIHf bull
t bull lt ~J~~~ __ ~
Norkr4irulB(198D) Advanculn UcrOfj~lpound~ltiOyVol 4 (AlItxlaquoIder fl Ed) PlerIw Press lcxIdon~ lIP~ 51-85 ~ l~l - bull bull
PJijllt~i At(fm)~mr~~JnI~1fqul1nQ JlalntlJ Riumh ~JnOrpnic toatJngs 2 192middot
)~J gt0 c ~lt -- ~ __ RogerS~ Ut (1968) ina ~(lnt~fbItliP4ili GI COnoslon ~a Technology)bull bull
Sl~iorr~ VP~Ranald~ Rctd~~ (1S80)_ ~ Experllattal Stud1aiRelate middotEffect or Cathod1c Protict1Oi withmiddot terta1nr1c
Coating Syst~ 12th Annualofr T8Id1na1ogy Co1fItHnC8 liauston TIIlt p 9-12
j
fIG1 sPct FromMon IT8TJCuZOI Int Lliu EOAX
FlG~ Fro TlTCu20Ar-4 Dy Field Ellposult I~ir
il
I c
R hi -00amp=7
-I I I I I I I I I f
iii
bull bull
bullbullbull
I J
J
I Figur 3 Thl chan In reltl rqpoIQnlof -ariOtimiddotjOmponents of the paint film with time Thl vtic~1 ampkiS unl till _n ofmiddotmt Illment per unit OJ fe 50all Cl Na
Cu StI Feeshy
PlistinllFe S7~J6 704 3~74 908
f
r e ~ ee~
middotie ~lt - ~ shybullt bull ~
t -~ l Cu~ o ~ f
t --- 3igt - bull 0 J Sn
cc
t ~
~
bullo
~ bull e
bullbullbullbull bull ~
bull ebull a QI 1- bull 0 40 431282 316
101 dmllIII)bullt bullbullbullt
hr 0 ~~bulla 5~16 ~~ -~~ i166 4S i83 8~1B
24tu 6~480 SD~83
111middot 1~1 OS6
~ 5114 6~69 -51 UIamp middot016- lt ~rmiddot
1d o Mea 818 21 021
~Z21 199 1~1Qmiddot ~Jgt2 058
14 d o ~~ 804 Q~B8~~~a~~t~middot 685
866 372amp 8amp4 OS _i~11[ ~~64
Tttile 1 A81UvlL pzQJJortlcw 0 vxl~oI ~~bull Jn 1I1Dtitctel lt-1166) end lqIlDteCt6 (O)middot~af~_~middot_lodEDf~ middotmiddotmiddotmiddotmiddotmiddotmiddotmiddotmiddotc
~ ~
_----I~
J
I bull
I
J bull tmiddot
7o CONQRESOIMTERNACIONAL
I D~CORRO$IOr(IAAINAEmiddot~USTACIONES~v c)
t 7thflfERHATlONAL CONGRESS
ON IARINE CORROSION AND fO(JUNG
~CONGRE$INtERNATIONAL DE~CORR~ON ~E ET QES~SAUSSURE$
i t
~ ~
bullt
t t
UNIVERSIDAD PourtcNlCAVollncio7J1 NiwiemIif~ 1988
(SPAN
SECcl6lfn
BiOIogro marino Morine biologY ~1i1orine
Theunpub1ishedJllBIlu8criptEl have been reprod~d asr~ielved from the
authors The SCientiUc COlIIIBittee take not respoi~ulbll~~Y for any shy
error or omssi9n
Laa ponenciasno han side publicadas y estanreproducidas tal y como
sellan recibido demiddot BUSmiddot autorea pound1 Conte Cientfico middotno se responsemiddot
bil1za de cualquier error ~ om1si~n
bullbull _ bullbull_ bullbull ___ bull $_ _ L _ - _ --- -__
SESSION II Marine biology
Friday 11th November
SURFACE PROPERTIES ON NONmiddotTOXIC ANTIFOULING PAINT FILM
K Nanishi M Murase Y Yonehara M Kishihara and T Hirama
MICROFOULING ON ALUMINIUM PANELS PLACED IN THE TROPICAL WATERS
N B Bhosle K Venkat and A B Wagh
EFFECT OF TRIBUTYLTIN OXIDE (TBTO) ON THE OVARIAN MATURATION OF THE PRAWN CARIDINA WEBERI
R Nagabhushanam B Indira R Sarojini and Mary Frances Thompson
EFFECT OF BIOACTIVE MATERIALS FROM SOFT CORALS ON BACTERIA ASSOCIATED WITH COMMON FOULER
Vitalina Mary Avelin Mary R Sarojini and R Nagabhushanam
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
t t
bull ~lS (3) ~ rflC~~ ~i~t~~l~~KI~ t~ldiIiiicyul~loIiih ~~Rht ~ )~- -~ - ~middotd 40 ( gt ~gt ~d lt1 t ~ Jt ~remiddot4amp tha paint trix cavity bottom eroded away reVe81~ng ore
t ~~t~ por~~ ~l~ P~]Y p~~8dby di~lutiaA~ ~~~~ti~Jr~gt~ta(l~~Siinll1cl1Ud~f iUiIiI reriatlGllbullt
bull a~~ ba~tid colCriAtloriccic~ GIl ~~~d ri -966 - ib~~~amp catQn1iti~~t~I~~(~t an ( ~
t the ~~~palnts (PIa_taiiI d~~h ~wh ~ n td have ~ CamplluJMitiIVc) ~~middottti o~lgtnalrf tlt of hIght
~ l=~A ~J~rW tYtles~ p~fiiilriQcOCCold to~Ja to jU-t ~jmiddoti~ai dlatClilS ~ir -jlati ~tbullbullent aNit ~ t ~~ th8middotf~ld~ies~~t~on~ ~ -966 prQ~~~(P14ta Iii~)Omiddotiliof~m1hiiHAiiipieat ere~ll-(piitir~ p-iot~iilans(PiA-iII~ end c) nbullbulltiledt dl~by1i~~ end wirinali1sheii ~cMI~bY thI tili
bull In t-~~ ~1i1nt 1oIIIeHfd~ ~m1IS (15OClind a t ri~r-dlatoa ~lor P~t dlataiOlonlt1anGIl ttw-966
t
bull ~
pr~t~ ~is~ih iion obitJauIJ A ~rot -=1 ~ ~ i __
reprasenttild on the 966 painted 1iUrfc1ncluicunljj ~ rwt
Achnanths Colonisation by bacteria 1n the laboretory bsed
bull eliperlMnt tollMct sbiUar pattern to the fild ~
bull bull Dilicueii
bull bull DUt 1Iidnt ~lona Mve dlttllrint ~~ ictian
It ~ JntM l1tenture ttt Iff yen __ JIll the
bull ~ bicins ani 111 ~t INchlnil (PhillIPs 1t73t EWene~ 1981
In ci1ntlHt tJWY ~tit ~Wl1c iIuCh as 1rbiltyl tirI
~ lite wAlII]ltiClr dlsP8tfu~ middotthiirsttilll~intaJn~ a
bull ~
lIIifoflll ampalCentraUon through the filII 10 thoIt biocide rel_ at tha
paint surfce 1fI$Jlts in IIIOre ditfusing ~s to repla~lj -iflJ~it
resulting 111 bullbulltNdy lr leechlng rata M1llat this IIIIY be middottruIi ot
tin ~ it was nOt true ~ ttw middotlBlf fOZWJlatiGn ua ~ bull - i - bull 1(1
ObSfvt1onundr SE clarly ~ tM t1l1 to be present
~ly lIS OIIIdle-llk parUc1~~t tn lUlfiice (Ito
IV) bUt usa ~ the dIpth attt fU-middot ~y~
If one considra the concn~rtlonmiddot chMge 111 1n ~111c COKICIl8rlts fro- the paint tU- i bullbullbull tMWei b~ ~ Si6t arid
tI-~t iron oxidt 1~ can bt (Fl_ 3 IrId TIibl8I) ~t ~ ase cona1derllble d1fftrenC8s ~ a Un decz bcdYMr ttw p1iod IIIIwraS lron~- an unt Ught inCr h ~ incqJper ~tlGn occUrnd _idly at ttw ~~ Gfb ~ period ~ out ttwn fu nPldly ampQII1n ~ umiddotirid II phast This IIIOUld bt tIlq)8Ctad It cantact l-=h1ng we occun1ng
IIltfIOS8d pertlcl of copper would lMch rapldly on t-n1on 1n - shy JIsewtr There would thlll1 ba bull J-a a1iltUy leu avalUba
i1 ~ ~
_tedal becfM uposed Thi tIOUld ttwn IIllCh Thla pulSing efttct
IIIOUld be expctad to -oth OIItlllith U- putic1 __ -lllible II tot 1~ GIl a on r-x _lao Thla teet 01 put1c1e 1tUng
can tit GIl the lIUrlace ClIlC8 it ~ a pan 1amp t
a 11aM bet in Plat I to IV IIaIrIg _ lit la_ II ~ 0- ~s _- PIate IW $llldl ~bull nshyot poRI flllNltttn Al~ tin ~ lou rete thtn I bullbullbull atill Ivwnce of bull al~t pulsed 1Itue rrlict tII1IIWd
elqlCtaG inIl ttwauUr ~ I I I
0
~ i
~ ~
~I J
bullbullt bullbullbullbullt bull bull ~
bull bull
I
bull t
~~
The same seqiHllle of cevity follliltloii (intttecaseofthetestjjltle
slot- shaped) fClliOileciby SecOlld8rYtlolesfotlllingin thll cavity
bottollis releasing further deep8rtox1ils (Plate XV d and g) As
cavities_re ProdICed ~ tiecaaIelnvadedby_ter~ onelilauld
expect 1ncreaa 1n1IOdha and dIladne lMls t the t1met
which the toxlM uw ~ Ths fCMld tCtHi eo (Flvmiddot 3)
~rttno the vi that_tat ~t1on otthe~ ~lY
il1Cl1l8S8S Idth UIiJ
Differences f~in toxin li~~to Ini 966 ill 8811P~ F ~ bOth Un end ~ lcMIr levels afillW
IIIIIlfI ptaInt(~1~~1ntha~~ ~~f~~ cst rklld with tbiIccpper ElJIc~l ta~Uona invalvinv
w pl8cId at a~ht~~middot3lO~lth Un cqJpel bull
(-960)10 ttW ~af this expei1tIIri1ts thIsiorelfJltdt -
that cqlp8r b8IiIId ~ IIiIOUldProc-ln f41ip1dly t thellllgllitiv
patentW _lied () as 1t 11 further f~1t ~ witt Un
III11ch is u-tat aqulllbd Idth
it If ttda 18 eo~ ~
~ )
frOll1 paintscould bl anqnced by an illprd current clithodic
protecUansysteipos1bly ahart_nino te 11fe of the paInts
antifouling ability Bacterial coloni Uon IIIU rtdUCed on the -s66 paint This
could have been due to unflSllOUlllble electrostatic cqrge on th surtece Th1a effect CIIl pley en IIIportant role in the t~of
bacteria to alrraces (lIIarsMll19E1O) As bacteria appear to IhiJw
greater toxicltyto copper (DMpsey 1981) it is Klre likely that
enhanced cqJper leacll1nl On the -966 AIIIPl c8used the recluCed
activity The IDlol8r toxicity ofUn to bacteria (as reported by
~ -
DelIIPsey Hle1gt or its lowes leaching rat was sUpPorted bythl
relatively rapid recolonisation of the slots by lIIicroorganiSlllS ~ftar thE inlt1al aurge of leachi~ Thailllpressecl current dto ~~ the oppoSite _ff~middot ~r on dtta colonstJon l1lI-1- in the
JIbcJrtory~ta larger rUDIIra f~ fit pro~td ~ther than uriprotected Pafnta
Ccnlus1one
1 tiild-a 18 presaIIUd ~ thattJn~ ~~
~llNChng notd1rf~ ~
2 TIleuseofCP~ ~CUIftnt(at) on1h1a ~fca41ng ~- ~
pa1I]t~ to ~_lAIIach1ngn_ at toxiN - -
3 leIJstaPidbectar1al ca1on1aatJon acaampmId an ~bId Pt1ntJ ~
4 JItore larger alcrofauJing (Oiat_ and PzOtoaoana) Gr~_
occurred on the protcbld~
I l
I
bull bull ~
mfEflpoundaS middot~-t9i _jgt- ~ _ _ ~sev M~J~ (1981) (cilonJzatiooof alUf~lng lnts by _rine
Jbull ~teda9g~1ta IIarina XXlV~ 185-191
- - gt 0 Ie 1 _ - _
poundV8I$i I~lf (1~P~h 1II8rin~Ugae and Foul lng AReviw ith
t p~l-i9Jrlr ~iiMdilto5hiP-fQUllfCiH 8ot~ ~r~ ~~~ XXIVbull lIPwl~J11A
Fotteth CN~R ~kk8nGB IWPIR1seJCiiiroaic1lihd ~e ~JIIth CII Offshore St1ucturu ElllS ~tjI~f~1Dt19i4
- - ~
UKC (1976) I~races ill ~W Ecology KIuverd Univbullbull middotraabullf~iddge ~bull C~~jII illt)Ia(t RC (1980) totinllsa Cattod1CPrOtect1cinbull middotmiddot8AtEmiddot~6haOCIlfif~ T~ p No 108 ~ 18 ~ 1IIHf bull
t bull lt ~J~~~ __ ~
Norkr4irulB(198D) Advanculn UcrOfj~lpound~ltiOyVol 4 (AlItxlaquoIder fl Ed) PlerIw Press lcxIdon~ lIP~ 51-85 ~ l~l - bull bull
PJijllt~i At(fm)~mr~~JnI~1fqul1nQ JlalntlJ Riumh ~JnOrpnic toatJngs 2 192middot
)~J gt0 c ~lt -- ~ __ RogerS~ Ut (1968) ina ~(lnt~fbItliP4ili GI COnoslon ~a Technology)bull bull
Sl~iorr~ VP~Ranald~ Rctd~~ (1S80)_ ~ Experllattal Stud1aiRelate middotEffect or Cathod1c Protict1Oi withmiddot terta1nr1c
Coating Syst~ 12th Annualofr T8Id1na1ogy Co1fItHnC8 liauston TIIlt p 9-12
j
fIG1 sPct FromMon IT8TJCuZOI Int Lliu EOAX
FlG~ Fro TlTCu20Ar-4 Dy Field Ellposult I~ir
il
I c
R hi -00amp=7
-I I I I I I I I I f
iii
bull bull
bullbullbull
I J
J
I Figur 3 Thl chan In reltl rqpoIQnlof -ariOtimiddotjOmponents of the paint film with time Thl vtic~1 ampkiS unl till _n ofmiddotmt Illment per unit OJ fe 50all Cl Na
Cu StI Feeshy
PlistinllFe S7~J6 704 3~74 908
f
r e ~ ee~
middotie ~lt - ~ shybullt bull ~
t -~ l Cu~ o ~ f
t --- 3igt - bull 0 J Sn
cc
t ~
~
bullo
~ bull e
bullbullbullbull bull ~
bull ebull a QI 1- bull 0 40 431282 316
101 dmllIII)bullt bullbullbullt
hr 0 ~~bulla 5~16 ~~ -~~ i166 4S i83 8~1B
24tu 6~480 SD~83
111middot 1~1 OS6
~ 5114 6~69 -51 UIamp middot016- lt ~rmiddot
1d o Mea 818 21 021
~Z21 199 1~1Qmiddot ~Jgt2 058
14 d o ~~ 804 Q~B8~~~a~~t~middot 685
866 372amp 8amp4 OS _i~11[ ~~64
Tttile 1 A81UvlL pzQJJortlcw 0 vxl~oI ~~bull Jn 1I1Dtitctel lt-1166) end lqIlDteCt6 (O)middot~af~_~middot_lodEDf~ middotmiddotmiddotmiddotmiddotmiddotmiddotmiddotmiddotc
~ ~
_----I~
J
I bull
I
J bull tmiddot
7o CONQRESOIMTERNACIONAL
I D~CORRO$IOr(IAAINAEmiddot~USTACIONES~v c)
t 7thflfERHATlONAL CONGRESS
ON IARINE CORROSION AND fO(JUNG
~CONGRE$INtERNATIONAL DE~CORR~ON ~E ET QES~SAUSSURE$
i t
~ ~
bullt
t t
UNIVERSIDAD PourtcNlCAVollncio7J1 NiwiemIif~ 1988
(SPAN
SECcl6lfn
BiOIogro marino Morine biologY ~1i1orine
Theunpub1ishedJllBIlu8criptEl have been reprod~d asr~ielved from the
authors The SCientiUc COlIIIBittee take not respoi~ulbll~~Y for any shy
error or omssi9n
Laa ponenciasno han side publicadas y estanreproducidas tal y como
sellan recibido demiddot BUSmiddot autorea pound1 Conte Cientfico middotno se responsemiddot
bil1za de cualquier error ~ om1si~n
bullbull _ bullbull_ bullbull ___ bull $_ _ L _ - _ --- -__
SESSION II Marine biology
Friday 11th November
SURFACE PROPERTIES ON NONmiddotTOXIC ANTIFOULING PAINT FILM
K Nanishi M Murase Y Yonehara M Kishihara and T Hirama
MICROFOULING ON ALUMINIUM PANELS PLACED IN THE TROPICAL WATERS
N B Bhosle K Venkat and A B Wagh
EFFECT OF TRIBUTYLTIN OXIDE (TBTO) ON THE OVARIAN MATURATION OF THE PRAWN CARIDINA WEBERI
R Nagabhushanam B Indira R Sarojini and Mary Frances Thompson
EFFECT OF BIOACTIVE MATERIALS FROM SOFT CORALS ON BACTERIA ASSOCIATED WITH COMMON FOULER
Vitalina Mary Avelin Mary R Sarojini and R Nagabhushanam
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
0
~ i
~ ~
~I J
bullbullt bullbullbullbullt bull bull ~
bull bull
I
bull t
~~
The same seqiHllle of cevity follliltloii (intttecaseofthetestjjltle
slot- shaped) fClliOileciby SecOlld8rYtlolesfotlllingin thll cavity
bottollis releasing further deep8rtox1ils (Plate XV d and g) As
cavities_re ProdICed ~ tiecaaIelnvadedby_ter~ onelilauld
expect 1ncreaa 1n1IOdha and dIladne lMls t the t1met
which the toxlM uw ~ Ths fCMld tCtHi eo (Flvmiddot 3)
~rttno the vi that_tat ~t1on otthe~ ~lY
il1Cl1l8S8S Idth UIiJ
Differences f~in toxin li~~to Ini 966 ill 8811P~ F ~ bOth Un end ~ lcMIr levels afillW
IIIIIlfI ptaInt(~1~~1ntha~~ ~~f~~ cst rklld with tbiIccpper ElJIc~l ta~Uona invalvinv
w pl8cId at a~ht~~middot3lO~lth Un cqJpel bull
(-960)10 ttW ~af this expei1tIIri1ts thIsiorelfJltdt -
that cqlp8r b8IiIId ~ IIiIOUldProc-ln f41ip1dly t thellllgllitiv
patentW _lied () as 1t 11 further f~1t ~ witt Un
III11ch is u-tat aqulllbd Idth
it If ttda 18 eo~ ~
~ )
frOll1 paintscould bl anqnced by an illprd current clithodic
protecUansysteipos1bly ahart_nino te 11fe of the paInts
antifouling ability Bacterial coloni Uon IIIU rtdUCed on the -s66 paint This
could have been due to unflSllOUlllble electrostatic cqrge on th surtece Th1a effect CIIl pley en IIIportant role in the t~of
bacteria to alrraces (lIIarsMll19E1O) As bacteria appear to IhiJw
greater toxicltyto copper (DMpsey 1981) it is Klre likely that
enhanced cqJper leacll1nl On the -966 AIIIPl c8used the recluCed
activity The IDlol8r toxicity ofUn to bacteria (as reported by
~ -
DelIIPsey Hle1gt or its lowes leaching rat was sUpPorted bythl
relatively rapid recolonisation of the slots by lIIicroorganiSlllS ~ftar thE inlt1al aurge of leachi~ Thailllpressecl current dto ~~ the oppoSite _ff~middot ~r on dtta colonstJon l1lI-1- in the
JIbcJrtory~ta larger rUDIIra f~ fit pro~td ~ther than uriprotected Pafnta
Ccnlus1one
1 tiild-a 18 presaIIUd ~ thattJn~ ~~
~llNChng notd1rf~ ~
2 TIleuseofCP~ ~CUIftnt(at) on1h1a ~fca41ng ~- ~
pa1I]t~ to ~_lAIIach1ngn_ at toxiN - -
3 leIJstaPidbectar1al ca1on1aatJon acaampmId an ~bId Pt1ntJ ~
4 JItore larger alcrofauJing (Oiat_ and PzOtoaoana) Gr~_
occurred on the protcbld~
I l
I
bull bull ~
mfEflpoundaS middot~-t9i _jgt- ~ _ _ ~sev M~J~ (1981) (cilonJzatiooof alUf~lng lnts by _rine
Jbull ~teda9g~1ta IIarina XXlV~ 185-191
- - gt 0 Ie 1 _ - _
poundV8I$i I~lf (1~P~h 1II8rin~Ugae and Foul lng AReviw ith
t p~l-i9Jrlr ~iiMdilto5hiP-fQUllfCiH 8ot~ ~r~ ~~~ XXIVbull lIPwl~J11A
Fotteth CN~R ~kk8nGB IWPIR1seJCiiiroaic1lihd ~e ~JIIth CII Offshore St1ucturu ElllS ~tjI~f~1Dt19i4
- - ~
UKC (1976) I~races ill ~W Ecology KIuverd Univbullbull middotraabullf~iddge ~bull C~~jII illt)Ia(t RC (1980) totinllsa Cattod1CPrOtect1cinbull middotmiddot8AtEmiddot~6haOCIlfif~ T~ p No 108 ~ 18 ~ 1IIHf bull
t bull lt ~J~~~ __ ~
Norkr4irulB(198D) Advanculn UcrOfj~lpound~ltiOyVol 4 (AlItxlaquoIder fl Ed) PlerIw Press lcxIdon~ lIP~ 51-85 ~ l~l - bull bull
PJijllt~i At(fm)~mr~~JnI~1fqul1nQ JlalntlJ Riumh ~JnOrpnic toatJngs 2 192middot
)~J gt0 c ~lt -- ~ __ RogerS~ Ut (1968) ina ~(lnt~fbItliP4ili GI COnoslon ~a Technology)bull bull
Sl~iorr~ VP~Ranald~ Rctd~~ (1S80)_ ~ Experllattal Stud1aiRelate middotEffect or Cathod1c Protict1Oi withmiddot terta1nr1c
Coating Syst~ 12th Annualofr T8Id1na1ogy Co1fItHnC8 liauston TIIlt p 9-12
j
fIG1 sPct FromMon IT8TJCuZOI Int Lliu EOAX
FlG~ Fro TlTCu20Ar-4 Dy Field Ellposult I~ir
il
I c
R hi -00amp=7
-I I I I I I I I I f
iii
bull bull
bullbullbull
I J
J
I Figur 3 Thl chan In reltl rqpoIQnlof -ariOtimiddotjOmponents of the paint film with time Thl vtic~1 ampkiS unl till _n ofmiddotmt Illment per unit OJ fe 50all Cl Na
Cu StI Feeshy
PlistinllFe S7~J6 704 3~74 908
f
r e ~ ee~
middotie ~lt - ~ shybullt bull ~
t -~ l Cu~ o ~ f
t --- 3igt - bull 0 J Sn
cc
t ~
~
bullo
~ bull e
bullbullbullbull bull ~
bull ebull a QI 1- bull 0 40 431282 316
101 dmllIII)bullt bullbullbullt
hr 0 ~~bulla 5~16 ~~ -~~ i166 4S i83 8~1B
24tu 6~480 SD~83
111middot 1~1 OS6
~ 5114 6~69 -51 UIamp middot016- lt ~rmiddot
1d o Mea 818 21 021
~Z21 199 1~1Qmiddot ~Jgt2 058
14 d o ~~ 804 Q~B8~~~a~~t~middot 685
866 372amp 8amp4 OS _i~11[ ~~64
Tttile 1 A81UvlL pzQJJortlcw 0 vxl~oI ~~bull Jn 1I1Dtitctel lt-1166) end lqIlDteCt6 (O)middot~af~_~middot_lodEDf~ middotmiddotmiddotmiddotmiddotmiddotmiddotmiddotmiddotc
~ ~
_----I~
J
I bull
I
J bull tmiddot
7o CONQRESOIMTERNACIONAL
I D~CORRO$IOr(IAAINAEmiddot~USTACIONES~v c)
t 7thflfERHATlONAL CONGRESS
ON IARINE CORROSION AND fO(JUNG
~CONGRE$INtERNATIONAL DE~CORR~ON ~E ET QES~SAUSSURE$
i t
~ ~
bullt
t t
UNIVERSIDAD PourtcNlCAVollncio7J1 NiwiemIif~ 1988
(SPAN
SECcl6lfn
BiOIogro marino Morine biologY ~1i1orine
Theunpub1ishedJllBIlu8criptEl have been reprod~d asr~ielved from the
authors The SCientiUc COlIIIBittee take not respoi~ulbll~~Y for any shy
error or omssi9n
Laa ponenciasno han side publicadas y estanreproducidas tal y como
sellan recibido demiddot BUSmiddot autorea pound1 Conte Cientfico middotno se responsemiddot
bil1za de cualquier error ~ om1si~n
bullbull _ bullbull_ bullbull ___ bull $_ _ L _ - _ --- -__
SESSION II Marine biology
Friday 11th November
SURFACE PROPERTIES ON NONmiddotTOXIC ANTIFOULING PAINT FILM
K Nanishi M Murase Y Yonehara M Kishihara and T Hirama
MICROFOULING ON ALUMINIUM PANELS PLACED IN THE TROPICAL WATERS
N B Bhosle K Venkat and A B Wagh
EFFECT OF TRIBUTYLTIN OXIDE (TBTO) ON THE OVARIAN MATURATION OF THE PRAWN CARIDINA WEBERI
R Nagabhushanam B Indira R Sarojini and Mary Frances Thompson
EFFECT OF BIOACTIVE MATERIALS FROM SOFT CORALS ON BACTERIA ASSOCIATED WITH COMMON FOULER
Vitalina Mary Avelin Mary R Sarojini and R Nagabhushanam
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
I
bull bull ~
mfEflpoundaS middot~-t9i _jgt- ~ _ _ ~sev M~J~ (1981) (cilonJzatiooof alUf~lng lnts by _rine
Jbull ~teda9g~1ta IIarina XXlV~ 185-191
- - gt 0 Ie 1 _ - _
poundV8I$i I~lf (1~P~h 1II8rin~Ugae and Foul lng AReviw ith
t p~l-i9Jrlr ~iiMdilto5hiP-fQUllfCiH 8ot~ ~r~ ~~~ XXIVbull lIPwl~J11A
Fotteth CN~R ~kk8nGB IWPIR1seJCiiiroaic1lihd ~e ~JIIth CII Offshore St1ucturu ElllS ~tjI~f~1Dt19i4
- - ~
UKC (1976) I~races ill ~W Ecology KIuverd Univbullbull middotraabullf~iddge ~bull C~~jII illt)Ia(t RC (1980) totinllsa Cattod1CPrOtect1cinbull middotmiddot8AtEmiddot~6haOCIlfif~ T~ p No 108 ~ 18 ~ 1IIHf bull
t bull lt ~J~~~ __ ~
Norkr4irulB(198D) Advanculn UcrOfj~lpound~ltiOyVol 4 (AlItxlaquoIder fl Ed) PlerIw Press lcxIdon~ lIP~ 51-85 ~ l~l - bull bull
PJijllt~i At(fm)~mr~~JnI~1fqul1nQ JlalntlJ Riumh ~JnOrpnic toatJngs 2 192middot
)~J gt0 c ~lt -- ~ __ RogerS~ Ut (1968) ina ~(lnt~fbItliP4ili GI COnoslon ~a Technology)bull bull
Sl~iorr~ VP~Ranald~ Rctd~~ (1S80)_ ~ Experllattal Stud1aiRelate middotEffect or Cathod1c Protict1Oi withmiddot terta1nr1c
Coating Syst~ 12th Annualofr T8Id1na1ogy Co1fItHnC8 liauston TIIlt p 9-12
j
fIG1 sPct FromMon IT8TJCuZOI Int Lliu EOAX
FlG~ Fro TlTCu20Ar-4 Dy Field Ellposult I~ir
il
I c
R hi -00amp=7
-I I I I I I I I I f
iii
bull bull
bullbullbull
I J
J
I Figur 3 Thl chan In reltl rqpoIQnlof -ariOtimiddotjOmponents of the paint film with time Thl vtic~1 ampkiS unl till _n ofmiddotmt Illment per unit OJ fe 50all Cl Na
Cu StI Feeshy
PlistinllFe S7~J6 704 3~74 908
f
r e ~ ee~
middotie ~lt - ~ shybullt bull ~
t -~ l Cu~ o ~ f
t --- 3igt - bull 0 J Sn
cc
t ~
~
bullo
~ bull e
bullbullbullbull bull ~
bull ebull a QI 1- bull 0 40 431282 316
101 dmllIII)bullt bullbullbullt
hr 0 ~~bulla 5~16 ~~ -~~ i166 4S i83 8~1B
24tu 6~480 SD~83
111middot 1~1 OS6
~ 5114 6~69 -51 UIamp middot016- lt ~rmiddot
1d o Mea 818 21 021
~Z21 199 1~1Qmiddot ~Jgt2 058
14 d o ~~ 804 Q~B8~~~a~~t~middot 685
866 372amp 8amp4 OS _i~11[ ~~64
Tttile 1 A81UvlL pzQJJortlcw 0 vxl~oI ~~bull Jn 1I1Dtitctel lt-1166) end lqIlDteCt6 (O)middot~af~_~middot_lodEDf~ middotmiddotmiddotmiddotmiddotmiddotmiddotmiddotmiddotc
~ ~
_----I~
J
I bull
I
J bull tmiddot
7o CONQRESOIMTERNACIONAL
I D~CORRO$IOr(IAAINAEmiddot~USTACIONES~v c)
t 7thflfERHATlONAL CONGRESS
ON IARINE CORROSION AND fO(JUNG
~CONGRE$INtERNATIONAL DE~CORR~ON ~E ET QES~SAUSSURE$
i t
~ ~
bullt
t t
UNIVERSIDAD PourtcNlCAVollncio7J1 NiwiemIif~ 1988
(SPAN
SECcl6lfn
BiOIogro marino Morine biologY ~1i1orine
Theunpub1ishedJllBIlu8criptEl have been reprod~d asr~ielved from the
authors The SCientiUc COlIIIBittee take not respoi~ulbll~~Y for any shy
error or omssi9n
Laa ponenciasno han side publicadas y estanreproducidas tal y como
sellan recibido demiddot BUSmiddot autorea pound1 Conte Cientfico middotno se responsemiddot
bil1za de cualquier error ~ om1si~n
bullbull _ bullbull_ bullbull ___ bull $_ _ L _ - _ --- -__
SESSION II Marine biology
Friday 11th November
SURFACE PROPERTIES ON NONmiddotTOXIC ANTIFOULING PAINT FILM
K Nanishi M Murase Y Yonehara M Kishihara and T Hirama
MICROFOULING ON ALUMINIUM PANELS PLACED IN THE TROPICAL WATERS
N B Bhosle K Venkat and A B Wagh
EFFECT OF TRIBUTYLTIN OXIDE (TBTO) ON THE OVARIAN MATURATION OF THE PRAWN CARIDINA WEBERI
R Nagabhushanam B Indira R Sarojini and Mary Frances Thompson
EFFECT OF BIOACTIVE MATERIALS FROM SOFT CORALS ON BACTERIA ASSOCIATED WITH COMMON FOULER
Vitalina Mary Avelin Mary R Sarojini and R Nagabhushanam
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
bull bull
bullbullbull
I J
J
I Figur 3 Thl chan In reltl rqpoIQnlof -ariOtimiddotjOmponents of the paint film with time Thl vtic~1 ampkiS unl till _n ofmiddotmt Illment per unit OJ fe 50all Cl Na
Cu StI Feeshy
PlistinllFe S7~J6 704 3~74 908
f
r e ~ ee~
middotie ~lt - ~ shybullt bull ~
t -~ l Cu~ o ~ f
t --- 3igt - bull 0 J Sn
cc
t ~
~
bullo
~ bull e
bullbullbullbull bull ~
bull ebull a QI 1- bull 0 40 431282 316
101 dmllIII)bullt bullbullbullt
hr 0 ~~bulla 5~16 ~~ -~~ i166 4S i83 8~1B
24tu 6~480 SD~83
111middot 1~1 OS6
~ 5114 6~69 -51 UIamp middot016- lt ~rmiddot
1d o Mea 818 21 021
~Z21 199 1~1Qmiddot ~Jgt2 058
14 d o ~~ 804 Q~B8~~~a~~t~middot 685
866 372amp 8amp4 OS _i~11[ ~~64
Tttile 1 A81UvlL pzQJJortlcw 0 vxl~oI ~~bull Jn 1I1Dtitctel lt-1166) end lqIlDteCt6 (O)middot~af~_~middot_lodEDf~ middotmiddotmiddotmiddotmiddotmiddotmiddotmiddotmiddotc
~ ~
_----I~
J
I bull
I
J bull tmiddot
7o CONQRESOIMTERNACIONAL
I D~CORRO$IOr(IAAINAEmiddot~USTACIONES~v c)
t 7thflfERHATlONAL CONGRESS
ON IARINE CORROSION AND fO(JUNG
~CONGRE$INtERNATIONAL DE~CORR~ON ~E ET QES~SAUSSURE$
i t
~ ~
bullt
t t
UNIVERSIDAD PourtcNlCAVollncio7J1 NiwiemIif~ 1988
(SPAN
SECcl6lfn
BiOIogro marino Morine biologY ~1i1orine
Theunpub1ishedJllBIlu8criptEl have been reprod~d asr~ielved from the
authors The SCientiUc COlIIIBittee take not respoi~ulbll~~Y for any shy
error or omssi9n
Laa ponenciasno han side publicadas y estanreproducidas tal y como
sellan recibido demiddot BUSmiddot autorea pound1 Conte Cientfico middotno se responsemiddot
bil1za de cualquier error ~ om1si~n
bullbull _ bullbull_ bullbull ___ bull $_ _ L _ - _ --- -__
SESSION II Marine biology
Friday 11th November
SURFACE PROPERTIES ON NONmiddotTOXIC ANTIFOULING PAINT FILM
K Nanishi M Murase Y Yonehara M Kishihara and T Hirama
MICROFOULING ON ALUMINIUM PANELS PLACED IN THE TROPICAL WATERS
N B Bhosle K Venkat and A B Wagh
EFFECT OF TRIBUTYLTIN OXIDE (TBTO) ON THE OVARIAN MATURATION OF THE PRAWN CARIDINA WEBERI
R Nagabhushanam B Indira R Sarojini and Mary Frances Thompson
EFFECT OF BIOACTIVE MATERIALS FROM SOFT CORALS ON BACTERIA ASSOCIATED WITH COMMON FOULER
Vitalina Mary Avelin Mary R Sarojini and R Nagabhushanam
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
_----I~
J
I bull
I
J bull tmiddot
7o CONQRESOIMTERNACIONAL
I D~CORRO$IOr(IAAINAEmiddot~USTACIONES~v c)
t 7thflfERHATlONAL CONGRESS
ON IARINE CORROSION AND fO(JUNG
~CONGRE$INtERNATIONAL DE~CORR~ON ~E ET QES~SAUSSURE$
i t
~ ~
bullt
t t
UNIVERSIDAD PourtcNlCAVollncio7J1 NiwiemIif~ 1988
(SPAN
SECcl6lfn
BiOIogro marino Morine biologY ~1i1orine
Theunpub1ishedJllBIlu8criptEl have been reprod~d asr~ielved from the
authors The SCientiUc COlIIIBittee take not respoi~ulbll~~Y for any shy
error or omssi9n
Laa ponenciasno han side publicadas y estanreproducidas tal y como
sellan recibido demiddot BUSmiddot autorea pound1 Conte Cientfico middotno se responsemiddot
bil1za de cualquier error ~ om1si~n
bullbull _ bullbull_ bullbull ___ bull $_ _ L _ - _ --- -__
SESSION II Marine biology
Friday 11th November
SURFACE PROPERTIES ON NONmiddotTOXIC ANTIFOULING PAINT FILM
K Nanishi M Murase Y Yonehara M Kishihara and T Hirama
MICROFOULING ON ALUMINIUM PANELS PLACED IN THE TROPICAL WATERS
N B Bhosle K Venkat and A B Wagh
EFFECT OF TRIBUTYLTIN OXIDE (TBTO) ON THE OVARIAN MATURATION OF THE PRAWN CARIDINA WEBERI
R Nagabhushanam B Indira R Sarojini and Mary Frances Thompson
EFFECT OF BIOACTIVE MATERIALS FROM SOFT CORALS ON BACTERIA ASSOCIATED WITH COMMON FOULER
Vitalina Mary Avelin Mary R Sarojini and R Nagabhushanam
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
J
I bull
I
J bull tmiddot
7o CONQRESOIMTERNACIONAL
I D~CORRO$IOr(IAAINAEmiddot~USTACIONES~v c)
t 7thflfERHATlONAL CONGRESS
ON IARINE CORROSION AND fO(JUNG
~CONGRE$INtERNATIONAL DE~CORR~ON ~E ET QES~SAUSSURE$
i t
~ ~
bullt
t t
UNIVERSIDAD PourtcNlCAVollncio7J1 NiwiemIif~ 1988
(SPAN
SECcl6lfn
BiOIogro marino Morine biologY ~1i1orine
Theunpub1ishedJllBIlu8criptEl have been reprod~d asr~ielved from the
authors The SCientiUc COlIIIBittee take not respoi~ulbll~~Y for any shy
error or omssi9n
Laa ponenciasno han side publicadas y estanreproducidas tal y como
sellan recibido demiddot BUSmiddot autorea pound1 Conte Cientfico middotno se responsemiddot
bil1za de cualquier error ~ om1si~n
bullbull _ bullbull_ bullbull ___ bull $_ _ L _ - _ --- -__
SESSION II Marine biology
Friday 11th November
SURFACE PROPERTIES ON NONmiddotTOXIC ANTIFOULING PAINT FILM
K Nanishi M Murase Y Yonehara M Kishihara and T Hirama
MICROFOULING ON ALUMINIUM PANELS PLACED IN THE TROPICAL WATERS
N B Bhosle K Venkat and A B Wagh
EFFECT OF TRIBUTYLTIN OXIDE (TBTO) ON THE OVARIAN MATURATION OF THE PRAWN CARIDINA WEBERI
R Nagabhushanam B Indira R Sarojini and Mary Frances Thompson
EFFECT OF BIOACTIVE MATERIALS FROM SOFT CORALS ON BACTERIA ASSOCIATED WITH COMMON FOULER
Vitalina Mary Avelin Mary R Sarojini and R Nagabhushanam
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
SESSION II Marine biology
Friday 11th November
SURFACE PROPERTIES ON NONmiddotTOXIC ANTIFOULING PAINT FILM
K Nanishi M Murase Y Yonehara M Kishihara and T Hirama
MICROFOULING ON ALUMINIUM PANELS PLACED IN THE TROPICAL WATERS
N B Bhosle K Venkat and A B Wagh
EFFECT OF TRIBUTYLTIN OXIDE (TBTO) ON THE OVARIAN MATURATION OF THE PRAWN CARIDINA WEBERI
R Nagabhushanam B Indira R Sarojini and Mary Frances Thompson
EFFECT OF BIOACTIVE MATERIALS FROM SOFT CORALS ON BACTERIA ASSOCIATED WITH COMMON FOULER
Vitalina Mary Avelin Mary R Sarojini and R Nagabhushanam
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
bullbull
lJi
SURFACE 1IIcftR~FiSOf~_-TOIle AHTIfOIlLHIG 11INTFIUI~
bull 1llani~ithramiddotYo~~8 II~lb_~1I1t1f TH~ra I~(~iDt ~~tt
ir~l ~lIfIH ~ ~1
p
AI1iT8ACT
As -Il) r~s ~Iellit~ 09t sliie ~nd is one of the
SOli proalsillll ___i s f Ilofttlie fttifClU~illl iltts But its -~ aacbaibullbulliof ple1IellflliC _ina feulillCS is Rot clew On the other
llaacl thfOitbosi~ f6flUti bsbeen aUcteci attention as It sialIar
phenoaenon to aerine foulines In order to Bake the relationship
betveen these two phenoaena clear proteins adsorption vas ex_ined on
arioU ailicona Several interestlnc coihcidents betveenthe tllO ~a were observed
bull I Intoduction
It has been incii(8ted that lov emero urfalt and also surface
ich is itt ~ifieran~ of enero have 100d lIintifoUlillC property t HZ) This is _ils explained ith thesurface eoerltY put it Is
not adequate to uplain for exale teflOft resin has I er surface
_10 than ilillconmin bill teflon resin MOWS poorerantifouline
perforuncethansiUcon resill
II1rinefouliftl phenoenonis theresult of adhesion and coarulaUon
l ofjSCOllIl Uquid3)4)5) In ihIs sense it closely reseables the
foraatioll of th~sis Reee~tb in the _dieal field sany art ficial
antl-thnMlboidpobr _terial have been developed ilne of ther II)lIt interlItine uterials possesses a unique surfalt strveture IIhlch is
bull collpoSed of separated bydrophobic and hydrophilic portions Such a surface ean be obtained by chaneine portions of NElIA (hydroxy ethy 1
aethacrylate bullbull hydrophilic c~nent) and styrenehydrophobic ~aponent)
t yenben thed_in size and the rieure of the surface a Iso dlance These
chllleea are pred_inatinr factors for oblaln1nemiddoteffevtive pob-r
lampirface Based on this fidines authors fnltestieated the relation
bull between 8dsorplion of proteins and asoftt of urine foulines on candidate
pob_rs by the use of various phase-separated surface
t bull t
2 Iiperient
(I) Materilrls
As hydrophGbie cooimt (A) IICthYpbenil~itOll~ ~ein
0IS1=1)7 anddi_thyl U~nllerubr (AlSt~) seleetel
As hydjUk c__nt (Ii) iHcotlmiddotaln_fa~~i~hfIQbJkYi~ amp101 II~ _lecte4 S- tructUlil 1 ~middotIti~t(A -
(Blare IIhova 111 Fieure t The _thyl-pbeftl Holane resi 111 151 toJ__lutilft the 4i-
ethyJ siloxane Ml inc 50S fast volatUeMJU tile silicon
resin aodified lIith polyethylene elycol in loDS N 1114 SiliCa reSi BOdeI flaquolIIfla vltiefl re coepollllOd c the eethy1shy
phenyl silOlt_ alld tlle silicon resin -fillet ith polyethylene 8beo1
and the sl licon rubber lIodel fonllla IIhieh were cOlllOllCd of the dillCthy I
siloane rubber paraffin vax and the silicon resin odified IIith polyshy
ethylene eyeol IIere prepared for follovine tests bull
(2) Sea i bull ersioll test
On 10 1300 II 100 steel plates epoxy Ale lias applied In t o
coats (100 aiClOllll lIZ) anelthen each saple vas applied to 30-50 icrons
thick_s and left for dryine for 1 veek at roo te-erature
Silicon resIII odel foraula and silicon rubber aodel fonula llere
i_rsed at Shibu Orido say fr_ Aveilst 1986lo flarchl987 and at
Yokosuka Bay froa flarch 1987 to January 1988 respectively The IIIIOUnt
of foUline oreal)ins vas val1Je4 at fouBnr percent every onth
U) Surface eneru
The nrface enerey as induced fra the cOlltact anele by usine
fowkes and Ow for_da The surface 5 cleaned off dirts and
lillC vbich adhered dudnemiddot sea i_rsioll and presented to the ashy
sure_nt
(4 ) Surface structure
The surfaoedried for a eek after application vas dyed vjth Osius
acid 6) and obserlled ith a SEII (Sannlne EIectro lIicroscope)
(5) Adsorption of proteins
Adsorption eaount as obtained by t~ Folin~s ~thod7) The
outline is as folls
1bullbull diaaeter glass beads hieh have about 2000 areasere
ia_rsed InZ4 Non Volatile solution and dried while shakinr After
that they were vashed in buffer solutiOft and then the air on beads surshy
face were coIIIletly evacuated Adsorbed proteins (Albulline rshy
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
gllthrin and Fibrinogen vere asured by lhe spectro scopic differene
using 770 nil tighl source
3 Result
(t) Si Iicon resin aocIel forilllla
The polarhy of the filas adsorption of proteins aDd asount of
adhered organiis are tabulated in Table 1 It was found that there is anoptiu cabination ratio (hydroshy
phobichydrophll ie cOtlponent (A) ioponent (8) ) for inihing the
fouHneaountvhich vas betveen8515 ~8012(t by vt in series
Surface energy dose not coincide vith this chaoee Polar coponent
of the surface energ) increasis middotsteadi b UP tomiddot 8020 and then rises
dra_ticab The reason seess to be the errct of the hydrophilic coposhy
nent
In cCOlIPany viththe increase of the hydrophilic coaponent the
surface structure Chances as shown in ficiateZ- middotPhJse separation beshy
coaes ot elearmiddotat the ~tio arou~ 8515 shoving the s_llest doain
size Themiddotdoln iUies are 08 Ja aDdmiddot 12 15 In at the ratio
of 8515 aDd 1030 respectively
It as found that there is an opUu ratio for each proJein vhere
protein adsorption calCbe kept at a 1nl_1 level These optilllJ 10shy
b1nation ratios beteen COlIPooent (A) and co~ent (B) are on 9010 for
Albuin 8020 for Fibrinogen and 8515 for T--Globelin respectively
These opthu colibinaUon ratios ie fro 9010 to 8020 are found to
veil coincide ith the iniaal foulinc allOunt See Figure 3 (~) Silicon rubber ~el forlUla
Tloe _ experi_tll as those for silcon resins vere carried out
vit~ sllicOR rubbers aA4 paraffin vax which are videly used as non-toxic
_tHouH lntll
_t __l1IIe f01lIi ~ surface ern are shollll 1ft Table 2
1ft ~Ii table 211~ tatlols relatiOfillbip vas not fouml either
The lip contf88t icroscope shooIed clear phase separation it is
_ in filMte 4 Althtgtuch the SEII holled vaglle phase separatioo
ilis not 8ssusebly appropriate to take the sase dyeing sethed [or
111008 resin odel forsvlas eperi~t Table 3 shows elcept for
the flatlol1 colltahtinc praftln v 10v adsorptioo of the proteiliS
corretaud to ampoed lIRti-fouling perfOfCaftCe Thollllh the foulat iOft
bull
containing paraffin vax shovs rather high adsorpti~n value its antishy
fouling perfor_nee belongs to the best group The result of i_tiers1011
is shown in Table 3
4 Dl scllsiOll
Although it is not veU kllOlIII about anti-throllbosis sechanisa by
the phase separation vith hydrophobic and middothydrophilic portions
following explanation is soseties adopted As there is an observation
that blood platelet does not chance their figure on phase separated block
polyser vlth hydrophobic aDd hydrophilic portions it is considered
that phase separation achieves soaethipg effective to restrain coagushy
lation of blood
The restriction of coaculation has 11 relation ith the adsorPtion
of blood platel~t which say be affected directly by the dosain of surshy
face structure Albuin bull r-Globline lind Fibrinollen are the represenshy
tative blood plasa proteins which have been investillated of their adsorpshy
tion phenosena on various aateriatsbull8) It b very interestinc that
if AlbUlline is said to be selectively adilorbed on the hydrophilic si te
and r--Globllne selectively adsorbed on the hydrophobic site to accua~
late the_elVes liakinc dONin vhiCh say restrain the function aDd coashy
gulation of blood
It is veIl kn~n that adhesion of sarine fduling is due to viscous
polyser liquid 9) As for ytilus the adhesive of foot (edulis) is
ainly coaposed of tyr~ine and takes quinon type cross-linking sechashy
oiss In aarine foulinlls as the first of all orcanic aolecular irrevershy
sible adhesions proteinsadsorption takes place and proteins forms
-conditionig fll-IOlll)12) As aarine organisss adhere onto such
a filII it is siailar to the thrOilbosis in the IlellIIe that the proteins
of God Iaaa 8Clhere OIl polyseric fUbullbull beforeMnci
I _ IiillPeliSlint ceplbrpar~itI UI bull the eb ~tIamiddot
hydri1ic ~ rie _ ~ti_ ~ bull ampOCMI allti-foulinc
perf_a and ~ a low a~1 ofmiddot -ceins It IIU~ t~t IIOt _IT IIvrfllice _rp llut alllOli9rpbOrogjo anillPOrt8llt flCtltllrBmiddot to
prevt rine hili I IRes middotThe fact that tere lire different sini_
val_ respecUgtveb fer UlMaaine bull T-GI bNt ampltd fibrinosetl __ that
the 4iffe_ II8l only cOme froetlieir daaractnbtics This ight lie
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
able to be proved by capelitive adsorptions which are coesonly used as
a useful ethod with edical lyHrs Although ~(fin wax worked effectively (or preventing rine
foulings the proteins adsorption did not decrease unlike with the
other ter181s Thu paraffin valmiddotmiddot anti-follline perfor_nee doesnt
rely on the proteins adsorption
II Refferrence
Applied lIicrobiolOIY 30298-308 (1975) U Outer S C
2) Baier RE PiocAnnu bull Contbull rc~liCaI Soc~ (J 973)
Biocheaical itcilllopIQ-Si~1 Rech CO_lini3) DevoreD P e~ al
80 No4 993--999
4) Lindner E and Oooley CA 3 International Conereu on llarine
COrrosion and FOIl1111amp 653--673 (972)
3rd Intemati~1 Biodecr Sybullbull 4174325) Cha~rlain (976)
6) Oshiaal and HaguilO T Karine Science lIonthly (Japan) 110 953-964
7) Low OH etal
8) BaskinAbulland Lyan DJ
9) Okano II et al
10) lIitchell R
II) 0e~1Ie IIJ
IZ) Baier RE
(I~8)
JBiol ChaJ93265 (1951)
J BloEdKaterRes 14 393 (1980)
Jinko-ZouU 10102 cl981)
Proceedings of OTEC Biolouline and Corrosion
Sy~osiubullbull (1978)
llarine iUolQampY 61 305-315 (1981)
Proc 3rd IntCangt liar Corr Foul 633
(972)
ftrdrophobic copOftent (A)( ( h Ph middotOIIe I ji Iimiddot
1SI-0 Si-OUSI- 0-Si-(OIIe)3I I I J lie IPh ~ OIIe nSI licon Resin
lie lie lie I i _ Sl-O-Si~O-Si-O- SI(OIIe)3I 1
lie Iff fie SUcog Rubber
ItxdroehiLtc CollipOMt (8)
i lie t 8 H
~i-O (CJf2)=-b-middotc~olHI I
lIed (it II Ib d1O-15 b 20- 25
Fiere 1 IIocfe1 pObliers used in the tug
Table1 rnterreJatlonsNpsof polari~ ofthe films adsorptionof proteins and amount or aahenkl organisms
1- ---- -1 Palarlty AdsartlCltt ~tiQmiddota Acth~dareil JlWCwitq(A~J br) IJ ~ and bilfllllClu
Wtl A I r
1 0 0815 7 222~urDlI1oQII eo 2 SUI 5 1131 O QtS ~I rIO
3 9010 12240 1118 0Il7 Q04 2
4 8515 Umll UO CI7S oOt Q
~ aolo 27212142 IllI4 004 o amp 75-ll SOl7U 131 ua shy 1
1 tOAD ~tH4 4IC 014 S
01 WH7dr~~(s)~IfIa~lOmt 2 AAIbumifr FJo~G(ilcmuilft
~
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
I
518020 ~~-Z=~~ A
6 --~--~--
10shy
12- 1~ S
IT f-r r
bull
bull _ l ~
1 -f
21-05 - t Fold or3191110
phase separation
4 185115 liJffJA Micro Pshy-r- VJi ~ separation
bullr~ shy
~ r~ ~j_1j _
+9
tgtshyerrltett ~
igtJ- ~b_ o
C1raquoi ~ o f=-
n =~
lio fshy i6ir
~ 1 ~
LZ shyo 0J
~ -GlOb ~
5 10 fi 20 25 Amountsof component(B) -CWUs)
Figure 31nteelatiOnshiPS between ratio of component(B) - adsorptIon of protein and adheslcinof marine organisms0 Albumin oFibrinogen ctor-GlobullnrAdhesion of marine organisms
W r I
~ ~ ~
f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
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bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
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l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
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L
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f Jt
I
Ibull I J J
~
Iftmiddottbull1ilHlhg r)(i ltJlIl1iflilJlIt paYlle placed In t~ trltp a I _~n
MI3 Rhode K Vellkll and AB Wagt NaUoftitl stt tule middot01 Oceanography Dona Paula
Gotlt middotmiddot403 004 flnd18)
ABSTRACT Hictofouling on a hlldn lUIII panels placed ill shelf slope and deep oceanic wbters of the Arabian Seb was tudied HlerolouBng biomass (as dry Weight andor organic carbon) w~s hither in the sheIt waters laquo 200 III as compared tomiddot slope (gt 200~it) And deep oceanic tater Mlonsst the orcanisms studied bacteria were first to appear on the aluminiuM surfaces and showed trresular srowth ~attern dUring initial stases of foulin (192 h) The number of foullnlaquo diatoms decreased as the depth increased rutudlll LimDQIhgfsect and HulsyU were the lIIost dominant dlatolllS Carbohydrate and liPids ~re the most abundant constituents In the touling material as compared with protein
1HTROJUCT ION
GoUd surfacet exrosed to ~awllter adsorb orgngt
-Uer such condlt1rJnd sUTtac~s phy 8n importunt role in
thf SUbl~H11i1I t Attachlllent of mierofoul1n organlslll~
ndudlnt bacterhl d lIttlftS Ilr(tOloa etc (Lret- bnd Net hof bull
1911 tJe~eha1tov and Ude) 1984) Adsorption of (ltgampnk
1IlIttei- and subseglll~nt growth of lIIicltoor~ai15ms on the
slldac~l ismiddot know as mlcrolotlling The develo~ent of
fIIhiroOOU~ iIIy IIIIWmiddot plJy an lmportampnt tole in IIttraditll
Ute -Itrva o-fiisny bntllU1i (Crisp and Raylanlt1 1960 Barn bull
t
~
T101 h-iiflf t~ _~middott~fftl~~~ ~Kh fl1i Imiddotnmiddotfn(t~ MAY
I i~ hmiddot~ _bullr1middotfilljot~~-~~~ ~f _~tl xmiddotlt~trrmiddot HmiddotU~)idbull
lltf nt middotn a~middot~tir li4 t i Ill middot1 r hmiddotw l)~
jnera~tn~friet1ltlIlre5l15twe~ n~r~jo1is t~13J andMJ
enhnnr- bPI~a Ie C()MOSt6f1 ((fefchaklV 001 Udy 19nofl
Very little information is avitHablflt on til develoPAlent
of mJcroflulJng on _1111 surfaces expospd temiddot ttypical marine
environmentlt Therefore in thi pap~r preliminary data on
the Inithl stales of lIIicrofoul1ng on alurninJuIII _tal
stlrfares have been presented and dlscussed
HATERIALS AND HETHOnS
Hlcrofoullng studies on alutninlum panels wr carded
(lut In ttl sh-lf laquo 200 m) I)middot-tmiddotlt I 200 ml lind o(lInic
Wiltmiddot 0f ~h~ IIIsbim ra dlld nf (middot-TIltgrlldmiddot middotlI f
ti rsfllrd vfssels middotBY lJy~)vHl_ ~rad l_(_ v fltlt)r tIfIYII
Th stlltirJrr cmiddotc(upled durim thsr (IU)5ltoS n dl~ln 1n middotl~
t nd Tlble 2
rr~rYrtJ~~ ~f ~~n~l~
AlIMinium plIllelS Hi x I fiI cnd re d middotod with
c)n~eotlllted hydrochloric altid IInd 1middotpeatedJ~middot washlt-d with
tap water flt)llltgttlfJ by distilled wat(-r finallY -ried in an
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
If middotimiddoth 1 111 4 ~ltEreltt HIt11 lI~lted or nllPclJI ri fj h~t~r lI fllHIl1 PIhC1~ 2 x 6 em 1 re f f~~~n~d ~1 1hmiddot~
J Thvr oC~~nJri sIifri ~middotatgtt~middot I wrmiddotr -I(f lt0lt1
~lId wr~ tlt ~rt 11 ltgtd hefI uSP lit th l~ ~t ti(ft5middot laifllmiddot(rmiddotmiddot1 inft v1 studi~d hymiddot d~rlolnr
Ill)ninlll J1lnll at ~ lItd middot4IlfI 11 (I11)g dlr e1l III-(rinll
I ~Ihmiddoty~nt rJf p~llftlli ~~ ~~
At eleej 5t-ali ns (FiR 1) alUllliniulII ranels wltr k~ft
I raquoJQ1~gl1 InJ ~1011~1J Ml1Ilv~h pf 1~J9f9uHII MtrbJ
SUlpended in th surface seawater ( - J ) f 10111 the oM p H1ltrofQUl1nc _terial fo~d()n the above penels -lit
Th__ (anel wer rltrieved after 3 to 6 h and evaluated middotfr various stationl and with tlllle and d~pth was seJarPEld usinjf
j MJc~fouling bioaas~ as organie carbon At these stations brush and filtered sltlIwater Samrls weI then diluted to II
surfare Mat~r sliMples Mere also collected and filtered known volulII~ and $ubsamplcs were taken for different
ihrnuch OPC laquoIus f1br~ 111teis to know particulate
I bioehe-lIicllil and biological anlllllsPls
or(ante carbon (POe)
o 0 Subsamples for diatOills were preserved with f(lfJrs A sing)e shtion (15 08 N 73 16 F) in thJ shelf
J lolution In laboratory they arl Howed to s-ttle for 24 h
wgtIter (Fhr 1) was coltcuplltd to study the inithl hiol(lIknl lton then cQunted IIn-d id1lfidmiddot uJlnr inertod lIticrrjseop~
M I--h1nI~ll sUutlmiddot of mlcrofulillP on alnmlnlum ~~elgt
I wUh rmiddotrJer1 Vmiddot H- lJnd depth Tim ~eric~ exp~~1m~nt WbS
Formiddot tllfl 8 11115 n-f dtmiddoty wtitht Q--eant- stt)(m (middot1 l ~Jtt~ M~ sertted ab()v excppt th t tt anelr w~r
bullI orliti ~~~ crtgtclI~tmiddott Jrl1gttlttA and lid II kno bull
INf ~tmiddot ~ It ioItNV1Jl dtJrin f1 rst 24 h 8hd tbreafhr v(l_ lttJtbs~J~s 113 fU~~ tnrJUlIh 1lilaquoHt
~ Olt t ~1amp 4~ Cmiddot 3 M anlf JrMJi~ ~tI1 ~~sft~~io-ht ~Jlfl
middottt--jk r~ 1-f(~S h tlw3 1II1crfou-Hng _8 atudhmiddotf - ltt rtmiddot(gtfl 1rIIIII Uptll w- iiUn~ middotumiddotmiddotmiddottest __~ t
al 09$4l ~~~lIItlgtmt r~~ _re d tlrot - t is IIIltJllthtt pSliOil at 10 2r JlIJ (If ltIIftd M 11 - ~
Qllll~ I lIIltl$IlQamp site t t-he above stetl lton JkQ-rlg IS lhefltIl-~tpIftYi~ ~il1)iwitat ~bullbullbull middott~A) aM FoU19-pJoeftC)
ltI~lod CQMailth 11middot1tfd rltreved 011 Harcenth 27 1986 relliJnhttowlV ~1I1 l~liriI~ 1V1b
4
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
shy
I shy
f
1
~
1 r
AsI- fmiddot~hn Il wmiddotr 1I~i tmiddotmiddot -rllrmiddot -1 I~TI
~ilutt~n~ f0r hnnt~ri8) ~n~ly~IR Tw(O gtr thn ltIi luUons
it~rlt rrJ middot~rI 1ust)rllc R P 31ater 11lpllc~t llqwts wlre
ftlt~rd throtllaquoh HUil1Ore middotfilters 11112 tim pltr siel- and
f11tfTl wr placed nn ZoBell$lnsrlne ntl1liellt 1(ar (No o
2216) plat~s The platEos were incubated at 20 2 C to
estimate of bacteri~l numbers
RESULts
)
Htrrf~ul1ng bitgt~ss filS orltnie elllbr-ni and the FQC of
the surface vater -gtIt eleven lI tation are sh=w~ in TablE 1
I1trrofoll1 hlg blltgt1ftllsS liS weil as JgtoC varied fro ltUon to
tiati middotn 111 gmiddotn~rlll 5tti(n~ occnpied jn hmiddott f wnt~r
700 III] 5h11 hlf(hr microfr)IIling thm thNI ltf tr wttr
P 200 m lind dor ltXtlnJ~ wl)t~rs fTU 21 Ithh
IIIICr(middotf IIU C Vt15 ~f1~fj but nf1 alw~)s asoclt~d ith tti~hr
por ltf the surfe Wllt1S rgtf the s~lf Itld ~1ltM stIU)IlS bull
K1erof( liOiTbh-WlIl$ (U tf n(middott gth Y rpr~ hb1( tilt f TfII~e
llh plte~ ItfId pthl n de VAt of he (~IJJ)tmiddot~ ttatiNr
(t-ampM~ 2)
A~nt the tetltmiddotorllIn~sills studied blltteria were th
Unto tp Colortlz~ the altiillillhilll ltlIJface and did lIt show any
delhi te jr~wth jgtlJIttern durlnthl2 h (1f dllmiddoty~nt Tah1-
bull I fol~ (11 1~l riH tr middot11 I~t 4 middot1middot 1
nli U-i r IgtIIlittnj w~rgtmiddotmiddot y ~ tmiddotmiddot ~ bull I f~ 11(1 ~i
middot~Jlptmiddott~t1r~ltl rarly chitnltd ltl~im 1 I Jlt~~ r 1I t I
h 1l1atmiddotrs Trj~~H~~lIl Mwj lt11 I r~ d~l I pM poundlt~tNrmiddotl-) ii3
vere nbs~rved in the lIIicro(ouJ inlaquo matedaJ of thmiddot tinl~
series
Ten different diatoms ele observed in the mitcr-huJ nq
mlter131 collected from dlfhrnt depths (Tamph1e ~I
tHmiddotZlltbiA vas the IIIOst abJlldant IroJp at all th4 dr-ds
T(Ital nUlllber of foull diatolll deltreascd as depth intnased
Of the ttn different iliatom fHent four lgtelrnd tt)
-tntnles and six poInnaJls At 10 2o 40 lind ~0 m dlth
thr~ to fol)ur typiS of diat11 ~r rct)rded wJler~al at t11
1ft 5~V~n typtloo~ (middotf dtmiddottom~ wr pp~nt [~1tI-tfmiddotr~ J rt C~
ill de lIIicrvfollJiIg matylli (11-c1~c fr dl rr rlt fnh
(IIf) nltlt b (IJrrl-d middotlu~ 1 rmiddotmiddotfmiddot~titj( rliffirllltj-1 t
h rd t~ hip
H1-r lltfl bl~ s n-imiddot it i~rhimiddotmiddot) plt~i T
vlIltd wUntj~ ana tHpth Thl -t fHtd _ A middot--t~rw r
hamiddot7t~ri~ Rftft dLlltltllll fI( -jt h middot)tiern liS n~ o~ltt 1liaf
l ni 11 shcemiddot ntt2 l t (If III e~()U lin Ll1 mt4
~g~ydTatlltmiddot _~e Wlt atgttlTlolli1lt III Hie rr-tlli n~
lIIaterl111 ~1r1ng ti_middot 5rt~-middot lIp1t lnrl Ill at v~ri)IIo
~
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
I t middot~rtI
I fIsCtlSrgt1tIN
fivrl paraMters loeh a carb(n 1I11))1I)11 ATr and Cltrfonate ~ont~nt fAftrlnlaquo and Taylor 1919 l1aYllck 1 Itl
1984) h~Ot transfer resistance (Chnackl~s 1913) and total
I dry vehht nf f(ul1n~middot Matertal (Burton and Harrey 191~ I
have be-cnused to qo~ntU the extent of blof(uIlnlJ We
I
s~ crtlnlc carbon IiIndor drY-weight contfltnt to ampssessthebull yt~nt of IicrofoJllhl 011 alttlllinlulII pattls with spac~
I ~ -
(Stations Ue and depth HlcrofouUnll was rehtlvely low
lh rlpe 1- 2~ 1111 andOceanic ater tis cOfllparfid with shelf
wlItoi-IS f 200 tn)ThlSiymiddot be dUiil tt the tfiffeNn-lI in the
hhJeoth~illteal andbiolodcal factOt-1I at th($1 thre
-n1 tvhllimiddott3 11ii suggested ttlil1 ~ IIo lyrmiddotsltr t ~rbullbull~i nd di-rmiddotlI tgtltyed ~n 1ri1p(rhnt tQle tn th d~elIgtiiIltnt (f
I IIIlc ~fmiddotIJln Oanshun H a1 IftR41- ThJ Js vhlen1 f rcoIII
th~ datlt prsented here (Tllble I ttgt o) We hhvp rtpQyed
I ldnlitlttJIit pn~1s rnil the- ship t iSUmMilt lIIi--tc-fNlllnR
1ttltlit5middot It lit1lerelore Poslibl~ that m~~rrJfulillJ w~s
(Hhlltgt VIrampstilitatd du~ to cohialillMtim frolll he sh J
resolll8b tv thUeffect wils slIall b~cau5~ we obJervdI ltQnddertlbie diffite~ceB in the fo1cr(fouling bi(lft85S In
sht-ll iUld slope i4aters HlcrofoullnlwlIs maximum ampt 2~ fn r ( Tllbll tmiddot ) This is 1n agr~eilient wJth NI Nriler
1
t-shy
h)~middotrtins rimiddotmiddotmiddot t Imiddot l~ 1 h n(middot~thr 1tfmiddottj t( Jf~fo- Pilll
rlaquol(n wherein mmiddot~imiddot lIl middotImiddotmiddotfUH~ WitS Obt3it-lt1 I
(wr amp Jforl ad ~( thlmiddotmiddot )I (WlIlh t1npubl r~~u) t~
VIIliOIlS w(1rk~r Itamiddot~ Ii~~~-=lmiddotlb d th~ developJnl sttP r
the primary sUllie film IHld prQlodcltl 5ucooli910n f tcmiddotmpey
lIIicrofoul1nu coinmunltiOl in tho tempprate ltnd sJh-tr(plca)
en ronlll~ntl fHit-hmiddotIJ lloli (riehman J984 Hhltp and PAmn
19804 Yanshun ~t a I bull 19134 l These studies lltugr lltl thai
blllcteria tier the fl rst rllJd SillS to bppear on th var1lt
surfaces lalted 1n
lIIicrofouHng IIIbter )
agreement w1 th th
seawater Our observation on the
frmiddotrm~d on aluminium tanl Is 11lt h
alJ(ve find i ftilI[S Jrre~ular ~rwt1
pattfTnS (If bact~rj amp all~ ltl1at(middotms I n l~rH el ~tS~middotmiddot~middot f
1111 c--Jmiddot foll] itll hltmiddot~ nl~ llmi 111 Ind ~ 5 middot1 tmiddotmiddot bull
~~u 1 t-lt~n~mmiddotl(middotn t~1 H i~t -t1 1 ~t- (middotrmiddot I ~ Y~fSltlr 1
111 19841 Thl1lt Irr~rtlll Irnwth pltttmiddotrn may 1gtlt h rmiddotmiddot1t
~f r~vorslhl IlttllthmlIt (middotf Imiddotlttltril to urf(~s hid rmiddot
relilnv-id dUf t lbvd 1 iI)middotSl lindc titt tr- ctmiddotcirn )0
prltotNrn HarIl-d I f~ ~l tIn l1arha I 1 HIlt I
After bactr1a) ~heslltn diat(fn5 Wfregt 1he fOcmiddotmiddotnil (rnUt
ot cl)]ltgtnists on the tilnlinj111 pane15middot Al ihobullIRhmiddot III Gns~ bullmiddotmiddotl I nves t1gatl cns of U mlcrofouUiil hay r~Ja1rl the
presence (r liiat~m~ in hr~ numbels ~middotw Itldled hm h~JI
8
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
7~
1 n fhv (r~UlI mmiddotmiddot 11ro ui lgtt hurth HUH
1ltmiddot 1 J Im 1 lin whimiddotmiddoth Iil] lIIInNIHimiddot lItr((
I i ~i 1( lI middotest r i I~(I ~ImiddotImiddot bull1 ~II billgt Id 1gtr lSIfl 11~ middotf
rt1 tilrrmiddot 11middot1 )i m I bullo 1983 roks~y cl 31
Igt~ In th1 prmmiddotmiddotflt~tldj in rddi-thn to lnnatltl
~Il flt~s __ r~middot niSI) rmiddot~I-dimiddot Of the vari Js dlllt-gtms
hmiddotmiddottt trl~~[if1Wl WFf~ U~ fi rlt to apP81 middotaftfitl 48 hrs c-n
J 11m 1niull panels f(1 IQwM by other dhttolll th
proett(Jftlinance o[ 1f~lliQ~ and tl0119ph2rl on the suhmrfaee
p3ngth fTableU indl cated the gtC()IIIlIIon app48rence of these
(~l)n151111 on hon-toxic slIlfl)ces exposed to vllrlolJs tpes (f
envronilerits r~
Bloch_leal antilysin (middotf the lIIicrof11111 shNled larft
amolnt ~r lipl d carbohydrat nd r~tetn whl eh lIIay 3ct-ve 8gt
fgtmiddotd sour fmiddotr t~lmiddot --- ) rt ~middotr Itlrv rmiddotf m
I middotfmiddot~lihp ()r~()JniIIIt
1I1tNgtloIIEroGJotlII~T
w ~yrrlts NIl i hcr th~fltS 1 [il rltertor (If th JnsH tIltmiddot
f(middotr hi~ intrfst tmd C(II5tMt hc-ourageJIInt w~ lls- thk
Usn Afgt IImiddotSoll~gtI 1 fgtlvilht and Shr1 NS [r)bhl t r
Ihmiddotir h~]r durIng h~ ltrnli(s
9
J(~ f ~rmiddotImiddotmiddot+middotf
IIfrinp k I m 1v I middot I~ ~ 111 fbull I limiddotmiddotIj flltJ tltl~ ~ 11 HI (tf(middot~n t11middotrf1~ t middotJmiddotrumiddot t~nv(1ttn (~~rjlllnt IIrmiddotIJJolIvjr~middot1 Mi-I jmiddotlJ ~ P H
Flarn8 II lrlO II rmiddotvj~middotw of 11111 ~tmiddotC Iff(- in th slttlemert and adhellhmiddot 1 thmiddot vptit f iM c)mmt)n 13 rn~cles i n l(lb~~1sl1J hi l~i l rgJ~1l) I~)~III~ F - Halt) JP Btl-Ill od (Ntlw Y-~rk 1IdmJmiddot~ Jr~I)
Burtcn D P and Hltrgrev S L 199 Control of fmiddot-lln~ organfsl)Il In estuarine ICQling water sYI1eml bv chlorine IItnd bromine chlorJde ~ll1lS1 ~cl- T~lttoJ 3 684-689
Caron 0 A and Seiburth J tI 1981 tlilttriblltioo (middotf primary 1011110 sequCnre on Ubrgllls - r1nffJrred p18$Ue Iubmerged in the marIne envlrNlmenf IIppl imdr91middot ~~~r~~lQl 41 268-213
Chllracklis W G 1913 Atta~hed IIlI~robinl -cmiddotwt hl 11 Frictional resistance due to microbiltl lll_St ~1t1 ~~ 1 1749-1258
CharcklU W G and Cooksey r E 1983 IH-fi 1m And liIiltrohiaJ fouling My ~PtJ H1st9H91 29 93-1middotHi
C(~k5ey R C(l)ks~) K E HHler CA PJul J H Rubin R W nt Whltr lgt 9114 Tho( Attachm~nt -middotf mi t7rrfmiddotfJi ng dbt IIIS In H~tUIl Pj-oJ~1middotmiddot~rl(orn~l(n l 1ttrmiddotrfjI~jpl IMr1 stv~y 1 1 h C~st~lr I lltmiddotppr r 17 bullbull ~middotmiddotII-fl V ~ IN
61n ltA I LtlId~~n
enpt ~ II 197lt H1ttltfFJu1in~ Tt r~) f rrimsry ft h flgtntling mnrln~ hltt~rho In 11middot(middot TIt r-l tntrp-li Cn IltPpoundttl$ Otl ~Itrjn~ C-nOSiN ~ill ~middot~I irC r--t R F IIck~~r 1 F Brown_ J R Of Palin W An(Ir~11 PI ~fI~ 6-~ Nrthw)strtl On I v fr$5 I
Crl~p n 1 Imd RlIhnd 1 3 Itlfi0 llfIn~middotmiddot middotmiddotf n Imi~ an( surf) t~ytlJr (middotn fhe 5~middotttlgtmmiddotnt middotf ~Imiddotrlnmiddot~ ~rlalims f~ttUll 11lfgt-lJl
Dnblts H flU] t( II HunlltcmiddotnmiddotJ 1 k~bmiddotmiddot rmiddot1 ~mith f i956 C(]Cdm~middottric methd f(I ~tt)rmi 11 i n f slIggtlrl A~~lr Cbr~ 2~ 3b~-3~6
bull bull
bull bull bull bull bull bull
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bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
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labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
bull bull
bull bull bull bull bull bull
bull bull
-------------
bull
Imiddotllt~h J tt- (-1rll1 H MhJ ln~ bull M fi llt1 Hutl)gt tomiddot C 1l1Ill middotmiddotl(ll bull JII middotgtr It trif~tmiddotI I
r1tgts dil~ ( biitroltJ IOf m bullmiddotrr-sl n at K~middothmiddotlfgt pmiddotmiddotlmiddot 1Wlit h~~ In l1(middotfmiddotgtHHQI ~fI~1 Q~IrQr~rJ lyml(~lIJ bullvlR II GrlllY PP 161-19 Ifd7h) and Wit Pa~fic H~middotrthwt Latbull Dept bull of Inergy)
lirlt-hakv SHand Udey t It UHIII l1i crofoul1 hi and Gmiddotrrr i 1n t1ulnt Bi~~uLqrtlpl ~n h1-udllaquolpHnlry 5Jlltly
pp 82-7 ed J D Cost-hmiddot ReT I per (ondon E amp F N SP-gtn LtdIbull I~b ) l 1911 Adsorbtlcn f an ltrganic f 11m at PhtUnIII1
seawateilnterhce ~ t1lJ[ middotStl 35 283-2IH
J 1 bull rY 0 H Rosebrough N J fair A L Int Randal R 1 Hgtl frltte1n 1Qe8stJrClQents wi th Folln-Neonol raf~Ylt Ith- QhflIIf 19326~-275
t I1llrshlJ] K C Stout Ft arid Hitchell k HI71 H(hanlm~ d h
bull Inlthl eventsln tle adsorption of bactltrla of
-- surfaces 2~D IUt~b191 68 331-348
I H~rRho) 1 K C 1976 Interf~ce in lIIiclobli1l ~1r-nmiddot
ffalltbdde bullHA lIarvtrd UI)lv fr~sll)
t HltlSlHk t A ~Crrlto-Q R 1 Wi Id E Wand fofE it It I l311 ( shy
pratlv~ effe-tll (f middothjrlr Ind cbJQrn~ c1iox1 hh lde rApt IIIfS fr tmiddotiy fmiddotmiddotmiddotmiddot H nv (Cnt roJ ~ot(~ ~~I ~ middot~~~-~99 shy
I Hltdbullbulli-J Fo Ilnd tJi~hlllrJO 11 11~4 rhlIIl~rmiddot-li81 middot11 1
lII~rtll~ ~urfJl~s ill Itaxlnc Bl(v~ri(-tImiddotmiddot1l At Hlt~r~~$-1rl n~ry ~ttJiy middotds J 1lt5t1)w k t f p I Lunckn Famp rN $PNbull tA1
bull middotrs-nr T ~ HatII1 Y and LIJIJ 11 C Ii llltl4 A lftuHal 1 dml ( Ilf~t1 t(lotclltll11throifn middotmiddotf ~ltwatr analysl INmiddot Yrtlo torf~ treamp ~ 61
Wl ~ ~ n f 3tWi (111)9(11 II 1 t4 I)ltt~rmi nn1 imiddotmiddot ltJf bimiddotmiddotnl~middot hy 6ic~i cat IItJltU cQnmmiddotni ty $1u~t1Jr~~ 1Jt~H1 l(tra(eJ 1 1111 plampqu( o( th- ml~r~toullnr fl(~ in 11middot~lrtl1~ llmiddot~d~rrj9rmiddotJ(p fn 1n1lflt1middotJpl Hn~lIY Co tu~I d 1 IJ Costoloil ReTh1gtl Pigt 8-14 I [mo bullbull r E F II pn Md I
IL l~middotmiddot-middotruL Y ~ II ~ ~ 1I~middotmiddotltmiddotmiddot~ y tH1 (itt bull rittf tttirI~middotmiddotfttdi~ tmiddotmiddot~lIj~middotm~middot in 7f Mtf
III I ffltilJlmiddot jn q rmiddot rt) bull IJlmiddot middotbulltignI~
~fIW ~-nUng pp 911 (AH~ GImiddot~c~
labl 1 K1ctQfou11nv bl on alua1_ ne1 place4 111 the urfce 18 at various hUonbull net particuiata ocvan1e cadlca (PaC) of Cfaee aten at theM taU
--------- -------~----------------Statton sU statioa lcMI1tlon bullLat 1oDO 11th thpth
( (---------_ ---------------shy
1711 15bull bull000 69bull4S0r 3100 t 5Sa ClSOO
3119 17 00011 18bull00 500 1 it4 569 3121 bull 3400 t 6629 U6al1900011 6150 1121 21 OOOmiddotN lIS 000pound 1600 1 68 1612 J72 20
bull bull000 n bull
bullOOOamp C8 1 102 100233
n 18 000 72 1501 19 1 245 393 3135 18 000 72 000 90 1 1211 0093
7 1800011 71450 80 1 217~4 aIMSo n4t 17 3O0middotN 73 000pound 45 1 17 n65
bull17 17 000 72300pound as I 1584 U4S 314 l ooe 72 000 n 1 11455 ]1- MlearopoundouUDIJ bla-U8 (ugC1h110 42) 8 POC(~-l)
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
--
--
-------
Vl ~ 1 tI I1 1f~ f~~~ r I j
t i r i ~ f 1j I ~ I~ lt~middott tr r 1 ~ I 1- - ro t ~ -nh t 1~Hj Hili bull
~~t
Table 3 Type IIDd number of lIIicrofoul1ngcUatoMon alUlaJn1a
~1 Fl t Ih
paelS placed at yadoubull depths at a staUon (15 0811 1316t)j - 1- I ~ I 1 ~
1 _______I-It _ ~It-~ ~ 1
SrKo Nbullbull of the l)pt W1~H A 8 A B oenus 10 as 40 60 10
-------~- -~----middotF~~s tlri 1 ( ~ It 41 fltl 1 - ii gt- ~ ~
fr=rti on gt gt
1 I~~~~ 6
(middot~11~~ r 14 4 1- -46 114 ~ bull ~ f 2 pound~dn~~se_2 2 Ar~1 d 1 ~~~ shy
rJ cmiddot s ~ktontel 3 W~5t(qr) 11 ~ P) ~~ L umiddot fjt~
1 ra~d t1f ~ 4 1I1ddl[i~ 4
f ru H I1 middotmiddotrq drl) Fr - ~i 1~1 ]I~~~ Irn 11 ~ 5 ~2 2
6 Thalasdothr1x 8 10 15 ~ -- shy
1 15Qr~l~~2
8 19 4~ze2 3 2 - 9 navicula 2 2 10 6 3 -shy
10 Nitzschia 118 591 189 10 --- 4
bull ---shy
Total 821 632 828 83 16
~ bullJ
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
Tab16 ~ Vlrlilttn In miclofolJl in and tts comrolgtit wtth 1m o 0
a~ a station It5 0~ N 7~ 16 El - _- - - - - _ - - - _~ - - - - - bullbull - bullbull - - w _ _ bull ______ _ ~ - _ - bullbullbull ~ ~ ( Ii
Protein lipl d 8ltVmiddotrir
(hrs) Cltlrbon Duration Organic GltIlb-hyd nILe
~ _____ 0-_
~
6 26490 ~~R it I I Sf -1~ ~ ti0 091~1~ ~
Il ~3 ~JYUH 0J~xI01lt -9 I I r12 ~
Pl Hi 11(1 R 1~xI018 PflIH If ~
18 fl6 4~0XI011111 II i 4 I 24 4
~B ~ 1 Gfl f middot1 J 15 17 flfmiddot ~6~xI0 ~
I r 116 t ~ 1 0~Rxi07 II If ~
1(11~1 o 12xlAHfl I I ~ ~ 7 1~ 7 113
~ p 1 11- bull fmiddot1~1 1 Imiddot
- - ~ I ~ IJ fl _~1middot1-1
It gt-1lH( 1 1middot1 1
1~l~ I I I ~~ bull I I ~~ ~i ~ I middot1middotraquo
~ fmiddot middotI-tmiddot- - l1~11Pi1middot_~nl~ )~(middotlk~middotmiddot r1
Imiddot 7 lhfttI~middot -)~
ID~ N tI lilto 0 0 ~ 0 ~ rt ~ IJ) IT _ Do Ji 11 r ~ It
~ jII o It i 0g tOrt aa a JI a
N N ~~ ~~~ g 0 w ~ ~ ~ ~ ~ e bull bull bull bullbull ft r IDW W (i aJ ~ WID 0 ~ ~
It ~ rt IT ~ S II
~ o It iii
~~ ~0 N N~middot lilt ~ 1~ 0 omiddot~O 1middot ID N N N lt _ q lID ID D go ~
~ omiddot trbull ID ~
It bull
CD IDOmiddot~ D I 0 pr g
~ fi ~ fl 8 j jj 1 D ~ ~ rt bull 0 ~O (Ij I If
D n S WmiddotmiddotW w W wl too 8~ 0 WmiddotN
0 W ~IO tI middot ~ ~ - it ~ ~I
W
Ii I
_ ---~----- - ~~ -
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
shy
~~13~____---~~]-------____~~
bullI bullbull
bullI to
bull E
bullo shybull
g
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
J 1
t II i shy U M ~ IS 1 bull 8 J Ilj I I j I~I-I Cf tmiddot1 bull -~ I- ~ I~ ~ I
al 11 f ~middotmiddotI ~ JI II I J Iiimiddot1 wmiddot1 -t r 1 middotIIl 8middot 1- a pound8
J~li tf iiI il iii i If IJ- 1i ~ J 1 I I 1 J i -_middot1 I ~ 1 I g
a i I J 1M 1- JI t iI ij t l - i J 1 yen I I 1 1 i it ~ ~ -l bullbull 1 Ifmiddot = I ~ 0 i it N ~ ~ ~fs1S I I 1 J i j I 1 imiddot ) til I i j J bull J ii- I - ii smiddot
If BI 1- 1 yen J 1 I 1 1 t 11 I i 8 J II I II jJ i I i ~ I 1 a Ii lJ hI I B amp i Ii eli 1
fj I0
1 g 1 I Ie I
-_ - -----~ - shy
-- shy
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
~- _---------shy-~--
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
~~
~ Iimiddot bullbull1Ii bullbullbull~ abullbullbullbullbullbullbull
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
middot ~tbullbullj
IE It
- a1 I f= i l ~ I =t Q ~ a Pmiddot I fD l~VI 0ltmiddotII 1middotmiddotmiddotmiddot1middotmiddotmiddotmiddotmiddotmiddotmiddot Jbull II
Il bullbullfmiddotmiddot middot I (I I Ibull 1 fl (1 I bull
~4 0 i-middotmiddot middotmiddotmiddot umiddotmiddot
A a_ f I- middotmiddotmiddotIt il -~lmiddot IIt I 1- bull ~1 II
IIlmiddotlJlmiddotmiddott~I bullmiddot e- f
J i ~ lr I f ~l f~i ~ bull
~ abull
lbl - 1
-tfect ofmiddot ~8ZO on ~ waqrof the PAwn lIlIi
CCnuol 2 hI1I hn 12h1 hJII 4ampII 15 4aytJlO dIlre
OyarIn India 50 ~ 43158 t 005bull 179
~(41)~_ eo ~ u t 10t~ l 2 2 341
a_bar 41a i2o~2t 01 (t 11)103 0_middot 15S
bullbulltun of aldClogt 1U0- 0shymiddotmiddotooplbullbullbull phlUomiddot pbll phU
8~ate of V1t4tl1o Pn- Pnshyoocyte Venio -n~l1o- -nteUoshy
gellla venle
- middot 21st 30881 19J~ 0506t231S19 024 0115
OJ tio~ middotIU 92bull~middot lo~ 069 12 0shy
0 O~ l~t tzotmiddot ~5 057 054 middot0bullbull- bull 0- BUOoo 0- ~l phil phil Phi
Pft- p~ JIIiw- bull 000001 TI~l1011~l1o-Y1VlteUo-middot IjItn10 110shy1jIt vento
veQfct
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
bull i
Dobull )
bullDo
4 bullbull
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
bull
-~4 uil
~
~i~
bull 1Imiddot
L ttshy 41
In ~ I~ 11111
Kt ~I~~I~
H~ ll1 i II
~I t- ID
tilbIbull c Jbull
-
I I
I t I I
INI
t
t-11 i K
u~~i ~ 18I
T
i 1-pound1c1
I I
I)1ft
bull
tilbull
Iigt
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
I
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
shy
EilECrO~BI~cnW KlEwts lROM -SOft (OBAIS ONMcrEaA
ASSOCIATED t(t1Itmiddot~R ~ULER bull
Sr~ vi1ALlRA~ioS~M~SrAWLIN MARY OS~M R$AAOJ1l11l and R~ RAampSHUiHAHAM
middotStioMaxytmiddota Coi1~ lut1COriD6 6280016 IDdia
bull IJepamprtlaitntof ZoolOgY Harath_$l 11DiveZllity6 ~8Dcp1b1ld _ 43100 MSbullbull IIID~
It -- shymiddotAbs~Ct
bull~ acterlsoc1a~d WLthBaiam bull PNtritemiddoten
iol~ed ud i4errt1fi_d bllSed on varloUil IIlI9lllhOlogiCal middotPhysiolQg1caJ Uld iilocbemical C1)aracteia bull 1bey ue ]Ibr106 ilcal1qenes aDdAerolnonaslUld all the eight bilCteri solates weJll9Ja1a D89ativebull MethyleDllcJUor1de extJ6ct
f)olll tb1e8 aoftcoralai Solenocauloa tortuoll1lll Echinooorgia CCilexmiddot-lndJunceUa juncaa middotwere foUDd to )) lnaeti1ll-bullbull Methylene chloriae extract fr01llSUberogorgta aUberOlfa prevent the JIIOt1le bacteria flOIa 9XOW1D9 oatbe tnated aur~ce
I X~t1cm
AeoD9fiZlltoE98Dis- to appear On asw-rged face
t
are bact8dwlJch y Ieee bullbullcurelyattAached afr u
little ODS bowbullIa~ Of ctolaquo9aoJS if ~ff8remmiddot bIIlve sUdlar attaC~Dt ktoetics 1he asion ofa prilllanmiddot ~
bull population OflD1cJ00r9aJds to_ioe surfaces bU been
bull stUdIed 017 (C~lland Mitcbell 1977Martll~1
1~78~00ni978) bull 1hebacterial f11111 then a~cts a
t diwrafi1lljWth protosoampatgae and fUllcr1thllt eventually
icoYeredl4tbJRU1ticel1ulaIIIIIQ~fOuliD9 (COrps 1910 a
Ma~~ket 1 )979)~ lhesl1- 1ayer coiIsi8tS of loose
alibullbull entra~d~ranicdlttr1t~a~dfiiiaI attaChed
bacteria with helMut _Chllnibullbullbull
~2~
Aatrong preference for 11_ surfacesbiss blen Obs4ltned
1nbarnacle~ and se4entary poIYChatea Such as Spirorbl bull
borealis (Knight JQneS6 1951) Barnacles JXrozoans and
oysters are alll9ng the IftOSt serious fOuliD9 proble6 oahip
bOttoms and piliD9B lhIpresence of -extracts fattyac1d shy
dedd butenolldeS prostaglUldinS aDd Dftrogen beterocrcles
(Baker and MUphy 1976 lurath I lbullbull 1978) with antibllcterial
aDtialCJlll antiprotcz0lll6 ichth]otoxlc and r~ other types
of bioloqical activity strongly _ug-geau that chelliical defence
is operative in gargoDfa and ore apecif1callY6 that prbIIIry
fila formation is inhibited in DIlDy of tt-ae anilllllla (Perkins
and che-siromiddot 1973 Tursch 1976 NaiDhe1llierl ibullbull 1977)
The present study was designed to illOlate and identify tbe
bacteria usociated with the cocon fouler bamacle and the
biological activity of aoft coral extracts on bacterial isotates
Material and MethoclS
Isolation of Bacteria
BalanUS lIIIIpWtr1te tlere collected frCll the bottora of the
ahip aseptically in sterile petridlah forbllcterial iaolationbull
APprox11lllltely 10 graas of BIlltnUil phitr1te _Upl was
aseptically transferred to a sterlle91 IIOrtarSl1ghtly
crushed with a sterile pestle and mi-4 _11 in 99 III of
sterile aged Ilea waterbull After givinglaquo fullabake with vartex
seril dllutioll$ were pmpared by adopting atandamprd procedtites
given by Rodina (1972) 10 lil of the inoculJll VU transferred
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
bull bull
~JI
-3shy
to sted1egibullbullbull petr1tUBheS and pOUr plBtecl he platies
~i~ted at ~OOllltelllperabu-e for ~4 howa he colonies ~~~diD tbepiltr1dtshes rec~unted ~ter 2-4howa and
t repsented on Cty welght bUls Par thilenWneratiol1 of
plllt - Yiabiebactedafi1lture medtumMampr1wae 2216 lqarmiddot 1 C4 (DlfcO~ntort ~tro1t MI) vuused I
Morpholgqicdoblervaticnsi OdttlnJ chlIi-acters lnciuc1ag
the appearanoe Of cultums the l_Of the colOnies thdr I J
outu eleyaUOft ttiiilsiuceacy ancvhether they are colourshylS 1Ihite Or othertdJle pi9Jaented wen noted
HIC101c9i1c41ObIitrvet1on Craa - ate1n is a differeDtlal
sta1I1qu1ril1g a p1IWIry stamptnandCOUtltlllr nUn he j
pdauyta1D i crystlll Yiolet vhich 1follO1jed by an
iOclOlutoo
SubCultures 1npeptone broth and on agar plates Sbceit
vu taPa-sible to exatdne 1n detail Illl the colOnies that
grew Oil the count plate a l1lllited DUlllber of coloniea (8)
wetemiddotiol~d from the pr~ plates an~were subculturd
both inpeptcne brOth Further purificaUODVU ccne by
repeated treaJdDg on the sea vater agar 1IIiIIc1a Of the same
C~1tlori~
Soft bull~imiddot2IaCt I gtSoft coralawere cOllected at luticorin
SoatheutCoUtof IntUa the f~sOft coralS were
SOlencic~u1~Dtoit~~ SulUoaor9t suberosa ECMnaAAmta
poundomplexa SAd JUneellajuncea Methyleae chloride elttrampctvu
-4shy
prepared (Scheme 1)
SCHEME 1
FLOW DIAGRAM OJSXlRAClION PROtOCOL
METHlLENE CHLORIlpound EXlRAClIOlii
Nt _lghtL1viD9 OrgampD1sbull
FlfIeze -20 bull C bullI I
_ LfOPh11isa UODFreeze cr1er bull bullI
middotdry Cllgallis - dry we1ght
- aM 100 _tbanOl (HPLC grad)middot bull _ COllect and tiltar
bullbullbullbullbullbull Crudethanol extract i
bull _ reduce by rotovapbullbullI _ add delolliseli weter to 9ibullbullI a ratio of 21
thanal I weterbullbull Aqueotm _thaaol
bullbullI) _ extract vith ~C~ 3 X 14
V01URe
Aqueo~ tbaaolCrude CH2CL extract
-~
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
I
bull I sshy middoti
middotmiddotmiddotlcnlw~jmi of extract wu ~~al1d _thyleoechlodde was j ~d by -c~bull SoUds were41bullbull01d l~lIea_~r and
~on~entrat1on ~ties _re prepare
ex Pour pite Jnlit~
ltIf
One III of thestrailiwaStratlSf8n-edlnto apetrld1Sh~
About1S 20 III of the nU1rlelit ampger_ltuIIIl was pOUred into
theptridi8hell at an ear lIearlDg ~lIIp8rature aSpetlcaUr
1liII 4Ulill eft rOtated in C1octWibullbull Aln4ilnticlockw1e dhec
tion for t~uqh adx1ngmiddot tfhentheavU soudff1~d fCUr
Mllctlcs _ftPlacedat~i~ points OIl theagu-ardace
lhid bullbullch4ic loaded With S u1 of each conceatratlon
RaUltmiddot
BD~rat10b Of Viable baCteria associated with Balanus JIIh1trite
be total bacterialpopulat1onin the pour platstlhich
are incubated at rOonI u-perature tor 24 hours ranged fran
48 X 103 9 to 88 X 103g on dry W1tight bllSi A total of
eight tratns wre iolailedampnd identified upto qeDeA level
With tba help of tbeKhe_ 91venbY Vdo and middot~Asfoe-_ (962)
and the resUlts ~re C~d with Bergeyl~ manual of Determishy
natl~ ~ioiogy (975)~
Id8nt1fi~ttJm of bacterIa
PoUrpi~8 which are incubllted at room temperature fa
24 houn ~obsemd for colony chllrillCtedtlcII Figure 1 to
3 Show4tf~rent morphol~iCampl cMJ~cte~Of the colony Some
are dUll wt4te or orange in colourm1le others amprecolourless shy
~
-6shy
Soma colonies are translUcent lOUrKi and regular RhiZOid
colombullbulluii ds() observedbull
ata - stll1p1nq
The _in g~pa of bacteria uecUsUnguiShed by
III1crDllcopiCal obSenatiOllS of their IIIOrpholcigy IIndlltaining
rec~onbullbull imt1ally in the gram- atampt_a prepaiatiobullIa
tt8 present obsenations all the eight bacterial isolates
are gr - negati Table 1 aho the idlmtificatiaa of
baCteria on thebaaisof IIIOrlao1ogtClll IIlcroscOpiClll oblleJll_
tiona alld lIlochltlli1aai ana1S~
Ant1b04ic SeOsitbitv Te8t
hi ampDtfbtotic peac111in diffuss tbIOurh the agar ~
OCCUP]1l9 a circular zone aroBId t ongiaa] spot 1Mgtowth
Of the bacileria WU seer in aU pllaquoces eaiept the circular ZOtle
where the antibiotic wu present StralOi 1 to 5 Showed
negatia reaults Figura 4 Shows the ~Dtegar plates
wfth stNirll l (A) lind strain 2 (8) Yibrto sp tnd1claquoting
ampntiblOt1c pencil1D resistance Strains to 8 showd pesitive
results Rutt18nt agei plate with AlealeaeDes s1) strains 6tA)
and 1 (8) bowing aat1b1otic senSit1Y1ty 111 show in Figure 5bull
Effect Of lIIIthylene chloddeemampc$ of soft coral_ on
bacterialmiddottsolatea I
1he -thylene c~or1c1t extraet 1oiaB40D tttedbc diffused
outwards fro eCch diac into the surroUDdiDgllV8l- 1i14
prOduced a diDdnlIIhlD9 gradient of C9llcentrlltion On incubation
I
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
lt bull
I
~- NcteJ1a ~W at arUofthe plate except thoSe udWld
thelDBter11middotto~chtheYere~~ri~itJ~an~ ~ wdth of
eacbgroiltb-freeampzehe of Inhibition I is ame~ure of
th~ir degree ofsensitlytty ~ ~middotlIIIiter1amplbull
No I zOne of lnh1b1tica t 1I8S observed inampllthe eight
strains when four different concentratiOils of methylene
~hlOrideextractfrom S~lehoeeulon toclUOllUm 70 119
Ech1ngogta ~~plexa 105mg and Juncella Juncea 205 1Ig
bued(Xl the wet weiqht of the Oxi9iJialllDimalmiddot other thtee-
cionCe~trat1on~_re 10 ~ 100 JJ9aridl JIg bUed on the wet
WliqhtOf the ori~irial aniJna1 (Fi9ures 6 _ 8)
It wU 1nteres ting to note the I zone of Inh1b1tica I 1n
all the attains when Suberogorq1a slbet-osa was lQaded ca the
disc tdthfour differentcoDOentrations i bull_ 195 1 10 1
100 ~g and 1 PC] based on the wet weiqht of the oriq1n81 ani_l
(FiguJe9) bull
DisCUSlonmiddotmiddot
middotaraal-Degatiw bilcteda pcedominate in the sea Moat
Of ~baCter1a ~ small rodS beinqdtherstramp1qht or helishy
coidal ~DCnilOt1i~ (ZoBell_nd Uphaill 1944) In ciur obsershy
vations ofbacteri~l isolates aSsociated with cOmon fOUler
Balail~middotDwhilli~ ~mc~ia1 pcpula~ion -is pred~l)ated by 9r-~9~iift foms 1hiee cento~ Ofhe~rotroph1c bacteria
Were1dent1f1ed~sedOnmiddotveJious IIIOrPhological phYSioloq1oa1
and biQChemlcalcheracterslheymiddot areVJbrO~ Alcaligenes and
Aeroinorias Hangi~9droPi pteparatJonsshow that COlllnOn
-8shy
sl~lI1D1 strains of ~ are 1IIOti1le OUr or1ent aqar pO~
plates show peak density of morphologically different strains
OrgampDiams tMt cannot be distinguished by mOrpooioqy
exhibit distinct diff~tence in thciJ biocbtlll1C~lmiddotreact1orur Bacteria differv16isly in their ability to atabo11se carboshy
hydrate to e thelll as a soutce of carboD ampDd enerqy lhII
present analtsia shows that i~ theatraiQ Aercmonu bull all
the other sn str~ns produce no6istectable gas ZObell
aDd UphUI (1944) stated tMt although ost _rine bacteria
can aillllate the siQIPle carbohydrate 11ttle laquocid SlId DO
detectable gas is pro41ced as a rule~ fba COrrosion enhanciag
effect of awfate reducsrs of the genu oaaulfov1bi-ioOb
cQrrodon ~oceaaes hUbeen attributed to the~r ability to
~hydrogen by hydrogeDUe act1v1tyfrca cathodic aitea of
corrosion cella (oerchamplto tt 198) I rson (1966)
presented direct evidence for cathOdic depolarization SiJillar
obeemtions were _de in the case of nitrate - reduc1nq
becteria vh1ch use M0 - as a teradnampl electron donor 1n our3
eight bilcterial isolates aeven ahow nitramptereduction
terosoigia dtrina contains caqxIDampI atructurally
related to the fatty acid deriwdbuteno11des vh1ih exhibit
antibiotic activity from ancep arid jUadalupensia (Sebild tz
and LoranCe1971) On site biouats indicated that alcOhol
extracts Of aeveralqorqon1anapo$as~ed considerable cytotOXic
ichthyotOJdc arid antibacterial activity (Jacobs i 198lgt
~ cHterpenes jeun1dn~deun1cin haw been isolated ampStile
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
bullbull -9shyImiddot
f~i I ln the 9Or9cOian Euni~a ~ frO
II c tdi1~ andBttnirespect$e1y 8cIt~refound to cytotoxi
a a9a1~i~~ NaUonalCaDceJ8Il3sUtutAil KB cell li Iha
it erild8 filxtraeti Ott~Btndni gorgOnian heMdmiddot cOnfined anti shy
iff neopluttc activity in MUonal cancen InsUtute lnvivo
lilt b1OUbullbull~A9amp1nstP -388 lllPhocytic (Alfred et l 1980)
Intblpnaeat tady bioact __italafra foUr oft
bullIt carals are neither highly toxic nor exaet111lke anUbiotlcs bull ThII crUdl _tbylne chloric)l extrtIlct frQIja sulerogorqia euleroea prnt the 1Iiot1le bacteria frOlll 9roviIll] on the treated udace other middotmiddot~lIIthylne chloddeSOluble crude extracts frOil
soienoeulon tortuosUmEchinogorgia complexa and Junce11 a jU1eeeit are n~ _ toxiciTheSe crude extracts shOuld be purified and
itls U8Ulllllc that the pur1fied fDIlterlal COuid be used u ait lIIIIanS of ptenUn9 IIl1crobial biofUm formation on the _dneit eurface1b1S tchnique-y provide proadaincJ result in cotltro-It hin9 slime rather thantbe toxic heavy IIIIItalbullbull
Referenceshyit middotAlfred J Js AM Helm DV and PollD9 M 1980 JllU1M anticancer agents 13 i4 - BIS Epijeunicin A
it _Ii cellblanol1cle frota the gOrcJonlan Eunicea mammosa
it ampa~rJIaD1HurphY v 1976 Handbook of Marine SCiencer
I copo1UI~ fro Mamprine Organiams 101 ICRcere8S
Clbull eUDd middotOhio 226 pp bullbull bull
sergeygte Manual of diterminative bacteriOlOgy 1975 8th Edi- tionBIlchllnanRE and NBGtbbonesEds Williamsbull
bull anc Wil1dn co BalUmore
bull
-10-
Carpe WA 1970 Ail eddpolyiacChIldcJe prodUcedby a Primary
pound11 - forml1l9 marine bacterIUm Dev Ind H1crobi~in40212
Corpe w 1978 Marine IdCrofodiIll] in DiEc heatexchJngers bull III ProcorEC BlofouUn andcorroeion -JIIpoBilllil ed~Rbull
Gray Department 011Z-91 Pacific Northwest Labcuatorymiddot
Report NO~ PNL - SA]lS Richa1~4 WA
Cunde1lA and Mitchell Jl1977 Mlcrobia1 aucicesloDOD a WOOden surface exposea ~o the sea IaternatBiOdetlarbull Dull 13 I 67 - 73
Gerchak6v SM andU~1 LR1984 MUlat BlOdeteriOCaton An interdisciplinary iIcly pp 82 -87 Ids bull Costlow JDbull aDd Tipper ac
herson WP 19i6 DirecteTicJence far the catbOdic cJePoshylarizaUQD theary of bac1tIriai ~oaoamp1on bull SCtence lSi 986-88
JacotRS White andS Wllaon L 1981 Fed Proc Ped JIM Soc Elq) atolmiddot 40 r 26
icD19ht - ranesEW 1951bull GltcJllriolSDeS and some other aspects of the ettin9 behampViour of Spirorbis J bull Mar Bioi Asaoc UK3i)(2) I 201 22
Maraball KC 1978 In water Pollution Microbiology 101 2 edR Mitchell ~w York John Wileyand Sonsbull
Mara zaleJc DS Gerchakcw S SM aDd VaL R 179
Intluene of subStrateCCMpoSiti01l Of -riDe ra1crofoiliD9 Appl Environ Microbiol 38 bull 987 - $IS
PerklnaDi and Chereazko IS 1973 1ha environmental toxieity of Crusin acetate UlSiD9 tetrahymena puriformla
aa a 1IIOde1 HydroblolO91ca 42 77 84
Rodina AGbullbull 1972 Methods in AquaUcitt~oblolOgy Univenity Part Press Balt1iaore
Schm1u 1 bull 1bullbull anei LoraDeeED 1971 chemistry of Coe~Dte retes XXI Lactones frClllll the 9OrgonianPterogog1ll guadalupenslSJ OrqCheia 36719 -721
I
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
r
~~~i1~=~~ o middotmiddot~rpallOi4sfro CQel~teratesiPp 247
l ~
R t
~ ~
~
t
-11-
- 29Gb PJ $cbiita~(Bcl~ middotMenM~ralPrQduCts Vol 11 ~a~C~s Nev Yon
UIIlo~$~ aniAal01t (962)Biiu Japan SOciety of Scientific nsamparlbullbull Vo128 Ro amp U1l33 - 11U
lnbel_r AJ Maison iIAbullbull Vai16rHelm D and Poling M bull 1917 Marlne antiClllICer agents asperdiola celllbranold frClllll the 1orgoxdana EUQica aaperUJa and tournforti JIetrabe4ion Liatt 1295 - i2ge
zoaell CE and UphUl C8 19U A list of arlae bacteria iD4u4iD9dIIsqdp~ClQa of 8ixty aev apecies BUll ScrippS
InlitOceampDOgrUQ1v Caiif 5(2) 239 - 292
j(
il
bull ~ I k
ftmiddot ttII too I
of
til
Ii II S tooI I
11
r
l ~
~
_1
f 1
~
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
Pour pla~s with nutdent iI9ar ill which bactedel colonies are developed at incubation middotpedodNote several morphological diffeientcolonies~
~~middotpi~t8 witliinitri~rit agar in which bacterial COlODiesare develooed
ltliiCiiaUotiperiOd Mote iieveralmorpholog1caldifpounderent(oloriies~
24 h
bull
at 4 h
Fi94
_____-_0shy
Fi93 Pour plate gtlith nutrient agar in which bacterial colonies are developed at 24 h incubation period Note severel ftIOrphological different colonies
-shy
Nutrient agar plates with straInsl CA) and (2) (8) Vibrio p showlrg antibiotic pericipinresistance
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
~
Fig7
4
t
lig~8
AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
gt~
AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
A
bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
~
L
~~~~~~~~~--~~
_~~~~ gtft~~-~middot~middotmiddot
tmiddot~
V~bdo p Nutdeilt agar plate showing no seilsitivity When methylene chloride extract of olenocampUlon tortucisum vas loadedbull
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AlceUqene spNutrienteger pii showing slight sensitivity when thylene chloride extrect of Echin9$2orqiaClplex vas loaded
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AlCUoen$ ap ~tdenteger plate shoving no sensItivity when methylene chloride extract of Juncella j~ vas loaded
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119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
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bullbull
bull
119 9 middotV1brlospMUtrtebt acuplate ahowlng ZOne OlIiiiilbltlonwhen methyleneehlodde extract f of Suberosavas loaded
2
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L