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Journal of the Egyptian Society of Pa.rasito~ogy, Vol. 36~ No.2, 2006
J, Egypt. Soc. ParasitoL, 36 (2): 545 - 558
IMPACT OF VARIOUS SALTS ON BIOLOGY,PHYSIOLOGY AND TISSUE SOLUBLE PROTEINS
(80S-PAGE) OF LYMNAEA NATALENSIS AND THEIRINFECTION WITH FASCIOLA GIGANTICA r\llIRACIDIP~
ByMOAMENA BAYOUMI MAHMOUD
Department of Medical Maiaco~ogy,Theodor BHharz Research1nstitute, P.O. Box 30, Imbaba, Giza, Egypt
Abstract
TIle effect of SLl hletl1~11 C()I1CCIl t r,lt ion S t)f ]11 clgl1es illlll SLli r)h ate(MgS()4), sodillll"l cll10ride (NaCI); 1)()t,lssill111 chll)ricte (K,Cl)~
ccllclllll1 Sltlph£lte (C~lS04) aTlel s()dillll1 carn()nate (Nil2C03) all.
Sll r\l j \trilJ ~ feCll11(i ity ~ L)'}Jll/(leCl 1](/1{t!(J/lsis i11 fecti on vV'i th Fc{sc'i{)!{!
(~(S{("11i(:ll 111iracidia allci the 'lcti\!ities of cc-rtain ellz,)/111es intissLle, 11()1110gcll(.lte of these SI1'lils vvere ev'llLlatecl. TisSllC SOlllllleproteil1S of the sn,tils were allaJyzed b)! SD,S-PAGE~ The reSllltsilldicated that Na2CoJ was ITIore toxic to L, rZ{lfllleJ1Sis that' t.}lC
other salts. Egg laying capacity \\',lS declined after two weeks ofCOlltinuous exposure to Na2COj (0.38 eggs/sllail/week) COllllJa
red to ( 13.85 eggs/snail/week) for controls. TIle lowest iIlfectlOJlrate was alnong snails exposed to MgSo4 (31.25%) vvith ~l
significal1t \!ariation fran1 controls (64.28%,). The salts supperssed cercarial production of infected snai Is (14.6 metacercariaelSllc.lil/stilTIll](lnt) at MgSo4 comp,lred to (59.3 metacerc'lriaelS11~til/stiI11ulant)for controls. The l11aintenance of L. 11at{llel1si~) illthe salts sollitions for fOllr suCCeSsi\le weeks reduced the totalprotein content in their tissues (13.2 mg/g) at Na2Co3 (100 ppm)cOlnpclret} to (2 t .47 111g/g) fl)f controls~'Tl1e acti\/lty of alkalinepJl0srllat~lsC \\/clS sig'lific(111tl)1 redLlce·d in treated snails t}1,ln COI1trois. T'he e)ectrt)l)}l()retic (lat~l revealed a COlnmon band betweetlcontrols ,lnd tll~,lte(l Sll~lils wit11 a Illolecular wei2:ht of 79,42 to
, ~
8().55 kDLt. Tllere Vlcre bands ill treated snails bLlt 110t in cOlltrols(41.2 & 4S kDa). A b'lnd WelS detected for snails treated \\'it}l
545
etlC 11 s~ll t hLll n (ll \v j th C()11 tr() Js ()r tIle ()t her tre at c.c.l grC)lllJs (28.6 XkD~l Llt I\T(lCl~ 1()().75 kDa at C~aS()-+ (\[ 2C)5 Kcl,t ~lt KC~J).
Introduction
Fascl()li(1s1s is a ChrC)11ic disease lflfecls anilllais. tIle 111(11n S()llrce () f II Ll ]11 ~ltl fase i() Ii 'l.~ is (E,l ~ S llazlye I (II. ~ 2()()2). Ii lll11 (.l n r~tS ~
.. ..i . ..
ci{)Ji~lSis in Eg)fl1l is (1 z()onotic IJrc)11Ie.Jl1 (H~lrid)' (!t (II., 1L)9~)} (.lnd11 Llllltll1 C~lSC S arc inc re.as in £ (H ,Jsec 11 (! r (II. ~ 2()()2 ). 1-6 \'IJ} 11.( {c(,
~ -
11(1{{I!('11Sis is tIle iJ11CrI11C(ji~ll.e l1()sl ()f F(IS(';()!{1 (~;(~(lllli('(1 ,111(1
111()st prC\f(lJCllt 111 tile 11lJrthern j){lrls l)f the Nile Delta (EJ-Aza7~~i
Lind SClli]]}l()rll~ 1983). l~lle C}lelllical C()J11P()Slti()Jl (Jf \\/ater 11"1
natllral \~"'atcr l)()(jies t"}c-lS <1 111 Llrkec! illflLlcncc ()tl sur\'i\'al .. ~r()\'v{tf1~ ~
antI fCClII1ciit), ()f sn~lil 1)()})ulatio11s. Tl'lis fact st1111Ll]atcd se\/e.raljn\.'cstlg,ltors l() eX]Jll)re tl1c. c()rrLI~ltiO]l het\V'C.Cll SI1Llils ;.JneJ tl1cCllt~l11ic~ll C()l11J)()sitt()11 ill tlleir \vater ll~ll)itals (EI-I-1i.1SS(Ul. 1974:DllSS~lrt~ 1<)7(1). l'\v'() t)'lJCS of Jlellrosecretor)' cells \vere ill 11)/
Ur( )III i Jl era] re·gl! Ia1" i() Jl j 11 IJ. St{llt?11{II is (B0 er ~lnc1 J0 () SC . ] C) 75).TIle It)\·v r~lti()s of calcillll1 to l11aglle.slLll11 aJld s()(fil.I111 t() 11()t~1~
S i lllll se\/c rc 1),1 i11 Ili bi ted gro\\t,th alld feCl111 (i i t)f ()f S11 ,11 Is (~Citl kaLlllli I-Iarris()ll~ 1976)~ Starvation of L~ Irl.fl1(·Clllt.!{{ ele\-'~lte(i tile hcl)~tt ()-I),lI1Cre,ls ellzYl11es' le\/eI (De:With~ I977 ). \1 e1(i 11ll iltZC11 an (lCalJerllS (1976) relJOrted that hel11()lylnph gll1C()Se C()llccntr~lti()11
llas ~l 1)()siti\le effect llpon \ritel1ogeIlesis <111d growtl1~ ~ltl(1 cel]L11~u~
ciiffercnti;'ltioll ()f felllale clccessory sex organs of L. st{ll~/l(l/is.
l'11c IJreSellt stllCi)! e\'aluatecj the, effect of Slll)leth~ll C()11Cel1
trati()flS ()!' 111agl1esillll1 sLllphate (MgS()-:l), soctiLtl11 chloricle (NaC)'). IJ()tassllll11 Clll()ride (KC])~ calcillll1 Sll]IJI1,ltc (Ca·So~) al1(i
S()C{ i ll111 Carl)()ll iltc (Na2Co.~) ()11 sur\! i \/ al alld egg Ia )fi ng c (ll)~lC it)l~1I1ci lJlfccti()Jl ()f L. 11(f!(llellsis. Also~ tl) cletect tile effect ()11 ]Jr()
tc i 11 C(lll tc Ill .. ~lC t i \! ities of anl inot r~lJl sfer~lse. ,Ill d I) ll()SP 11~lt,lses ell
Z)!lllCS itl S11ails tisslle )1()l110gcll(lte ,11lCi tIle cllanges in elcctr()pll~
aretic pattern ()f tiSStlC s()]ull1c l)l"oteins of L. l}t/lll/f'llSis eX]losedto fOllr sLICCessi\,~e v/ecKs to saIlS Slll))elilal C()llcentratiollS.
Materials and MethodsL~'<~/rl17(1 (~(l 11(lf(11ell s is (6-8 111111 ill heigIlt) Vlere Iaborator)f t)red ~
,111d Jllailltained in decl1lorillated aer,tted tape \\I(ltcr (10 sllails/liter) at COllditioll r()()lTI (24+ 1nC). TI1ey were fed O\'ell (lrlccl let.-
tuce leaves and hlue-green alga Cl\"ost()(' lllUSC()flf}}l}. Egg 111asses
\vere lsolated and \\leekly counting the eggs nunlher.The salts sodiulll chloride (NaCI L potasSiUI11 chloride (KCl J.
sodiull1 carbonate (Na~Co:)~ ca]clunl sulphate (CaSo~) and lll'-l
gnesl urn sulphate (!\1gSo4) \vere obtai ned in pure fornls T'hreereplicates \\Jere prepared for each salt.. each of ten adult snLlll~,
(6-8 111111 in height) was illll11ersed in a liter of the salt con
centrali()ll. l~hree replicates of control snails \vere lna\ntLllncdside by side under the sal11e eXllerilnental conditions in dechl()rlnated tap \vater. Fooet \vas added to daily~ dead snails \verc reITIoved daily and the salts cone. \\/ere changed \veckly. The nu
llll)er of eggs/snail (:t\1 x) and survival rate (Lx) \vere calculated
\veekly through the four successive \veeks.~')nai}s (3-4 11111) jn height) \\/ere eXl)Osed to F. ({j(~{{ntic{l 111i
racidia obtained fronl Schistosollle Biological Supply Center(SBSC). Each snail \\/(lS exptJsed ({) ~-5 1l1jracidia (J\1alek~ ]980:Moens~ 1991). l-'en snails \vere kept in Petri-dishes contalJ11ng5() ppnl of each salt. The salt concentration \vas changect tVv'icc
weekly. The snails \verc fed on blue green alga only and Inaintained in the Jahoralory at 22+ IDC. Plastic sheets \vere placed\vith snails after a 1110n111 franl date of infection and metacercarlae \vcre collected fro111 these sheets in each Petri-dish. Shedding of ccrc~lriac \V,1S for 72 hI" 'and lnetacercariae appeared ashrown l)oil1ts on the plastic sheets. Snails l soft parts \vere dissected ()ut 1'ron1 she Us by gentle crushing the tIssue vv'ei ghted andh01110genizcd in 1.5 1111 ice-cold distilled \vater hy a glass h01110
genizer for 5 Il1inutes. The h01l10}2;Cnates \vere centrifuQcc.! for 15~ ~ ~
1111 nutes at 3()OOrprn and the fresh sUl1c.rnatants \\'cre used fortotal protein and enzylTIC assays.
Assay: Aminotransferses Cllzynlcs (aspartate anlinotl'ansfcra
se (ASAT) and alanine anlinotransfer~c (~t\L}\'T') \\.Icre as~aycd
(Reitl11an and FrankIe, 1957). Alkaline phosphatase (AlkP) andacid IJhosphatase (AcP) \vere carried out according to Kind andKing (1954) and Moss (1984)~ respectively. The Biurt 1l1ethc)d(\\!ooton'\ 1964) was useel for the (Jeterlnination or lota) protein.The parUllleters were deterlnincd spectrophotolnetrically and all
the Kits \vere purchased (Bio-Mericux Co. France)" except acidphosphatase Kit (Qui1l11Ca Ctinica, APLICADA~ Spain).
Electr()1111()rctic LlllaJ)/sis \-\IL1S c.i()ne acc{)r({lng t() \.'1,-te(j~1 et (,I.( I <)X4- )+ [)cterJ11 in~lt i() 11 0 f the III () 1e·CLl ],1r \\/eig11 ts () f sell ~.tratc(il1~trlcls ()Jl tIle f11cth(){J t;111111()y'eci !)y' !\/l()star~t el (II. (]()()()),
Statistcal (U1{tl\,~jS: Datn jl1Clu(iin~ l11can \'aILJt~s. starl(l~tr(i (lc\'~~ ~
iat i() 11 a11 (1 Jl ~ \"' till.l CS \v ere c ~1rric (l () Ll t (S 1.r ike 10C) 1).
Results and Discussion
I-J. ,./(I f({I(J , Jsis \verc c()nt~i nl.l () LIS J Y' exP()seel t () cl if f'c rent S(l ] t ~ (() r
4 weeks. "fl1c sllrviva] r'.llc' (l~.\) as <I fractic)11 ()f ()nc (1.() =1O()(J;(,). IllC(ln l1l11111)er ()f e.ggslsllajI/\\/ccek (]\:,1:\) tIle Ilrc)lluct ()f
1-,\1\.1\ ~tll(1 the SlflllJl1atic)Jl ()r L\I\1\ (R l)) tllrC)llgllollt 4 \vcek,~
(Tab. I ). At the 4lh week survival rate of control group \\iaS
11ig]ler tllall th(),~e r()r sn~lils CXI1C)Scci tC1 t}1e e"xI)erit11ent~11 salts
IJcillg X()\k" C{)1l1})arc'(1 t() 33(/( :Jllci 1(-)(}(I f()r snail gr()l.ll)S CXll()secl
to SOC) 111)111 l"\~lcI1 frC)111 KCl <111(1 (~aS()~~ resl,ccti\/c):,r. l\!J()re()\'er~
110 Sllail sL.lr\/ivcc! at S(){) })f1l11 Na2(-"().~. l"hi~ incljc(ltecl tllat Na2
Co_~ \\/(\S lll())'C {C)X1C t() l.,. ll(ll(l!lJ!lSis sn~li1.s tl1an tIle ()tller testeclsalts~ C<'11111)~lritlg the effect ()f ]'<aC'l ~1t1l.1 I\ra~(_"()~ (5{)() llIJlll)
S}10\VCll tllat tile Ili~2J1 tC)X1C effect ()f Na1C'1(),~ \V~lS CillC t(J (_~t)- ~ i(}ll~~ - - ,
rather than Na+ ions. The exposed sllai Is to 1DO ppm or ?\Ja~Coi
could 11()t SLlr\lj\,c Jl1(Jrc tllJrl tllrec Sllc(~cssi\:'c, weeks. l~lle rcsLilts
agreed with Mostafa (1997) and. EI-Hassan (1974) who statedth<I t. B. {IZ (~),~.(Ill (I J~i'1 II (1 id 11 () t tal eratell i g h c()11 Centr~lt10]1S () f N ~l")
~ ~
C()le tIle dccre~lse. ill L. 11(lfcl!(111Sis })rOdllctivity' (R()) se,ell in tilepresel1t \\.!()rk in tIle C,1se ()f N~l:2C()J cllld C~lS()--l 111 a)" pLlrliLll1)/ lille.to C()llSi(ler~ll)l)'r lovv' SLlr\ri\lorslli}) (L\) of sllaiIs at tile salts C()Jll
p,lrecl t.() c()ntr(ll ()llC-S (111(1 (M.\) \/~lllles ()f SLlr\?j\'cci sl1aiJs \verever)! ]ov./. TI"le R() \/ailles \\Jerc· rcc{)rc{e.ci f()r snails expc)seej tel S()()
alld ]()O PI)111 ()f N(12(~()J 11e.ing 1.8 for eactl~ 7.() fc)r sJlail~ eXI)()
sed to CaS()4 (500 ppJll) witll a reclllctiOJl rate 95.2(;~" & X{J.JS(;~ ~
re-specti\lel)! frtltll control. The results Sll()\\ieci tll(1t i'V1g:S()4 ()()()!)}jlll) )la(1 the le~.lst harlllftt] effect ()Il L~ l/(tl(I!{)/lSis ill C()11111,lrlS()11
\\lith ()tllers vvhich s}l()\vecl Llllig}l SLlfvival rate (63(;(')~ Re~arLilll~~ 'L 1..,.
Ro \/~l]lles~ rcsLI1ts revc,llect tllat Sll,-llls CXP()scct t() MgS()4 \VLL~
40.4 ~lPl)roxiI11atcl)! as c()tltr()ls. DC-\\'llll (1977) rCI)()rtc(I tllat
IO\\J ratl()s ()f caiciLIJ11 t() l11(lgnesiLl1n~ s()liillIll ()r P()t~lssiL1111 se\~e
rely inllillitecl gr()\vttl & feCLltlciit)-, (Jf L. fl(l(tlleTlsis. Rel(lti()IlSllilll)et\\.leeJI \\/ater cllelllic~·l1 C()Jl'lI)()sit.i()n & rCIJr(")clllctlC)Jl \Vas
'-rable I: r:ccunJity 01' I.J_VIJUIUell l1(ltlllensis after 4 "leeks of continuous exposure to different salts for foursucces~ive \vcekS
,._---21~d week "'1(1 4 lh \\'eeKSaIl.":; in I ~~ wcd~ 'I w('ek Ru 1(, l)(
-[lPIl) L\ 1\1 \ L,I'vl, L\ h1, Lxl\1 \ L\ tv'l \ L)\t1\ L\ fv1 \ L,-M, (~L\M\) Chang.('
....----~
Coni rot U.lJ7 9.2:.1 X.l)_~ 0.93 I J.90 12.89 o.lrl 7.~U 6.70 0.80 12.lJ2 IO.J3 Jg.LJ
[\lgS{ )1
_~{JO (LYJ 1(~. j; I 15.)~ O.7C) I ~.-l() 14,01 0_76 S.IO 6.10 0.63 7.47 4.70 4ll.-l -t-3.XS----~ '~ ..~f---
l'\i~lCI
5()() l.OU S.6~) 0.63 O.7~ 22.)U 16.42 O.6J ~.J{) 5.20 0.56 8,64 4.84 35.1 - t). ()l)
KCI
)l)(J U.70 0.23 0.16 O.S6 K.OO -L--l-S O.-i:i 21.){) Y.20 ().3~ 8.70 2.37 [C)_X -:)6.')
C~S().j
:)U(} U.7h U.)2 ll._)l) O.3b 1I. ()(} .~ .t)l) 0.26 9.40 2.40 0.16 ~ .20 O.X3 7.6 -~().J)
-~---
N~l.'C()~
IlH) {l,l.») (). __~6 n..-=;) ILb's <l.-iU ll. )~ O,2~ :1.UO I.JO . 0.20 U () 1.X -Y5,24
-, .... , --I----
l~~;;o.X~ 2.S~ 2.2u O.t-lO tl U ~L65 0 0 (L~O () () '1 '") -94 ..:"+
ILX3 2.12 1.75 O. --tU O.JlJ (). l () () 0 0 {) 0 0 LS -l)S.2.:.\-I I ----.1_
).+LJ
l"able 2: SurvivaL infection and cercaria! IJrc)ductl()1l (n1eta
cercariae) 1'1'0111 L. nUlo/ens;s exposed to F. (![ijt!.o!ltic(l llliracidiasllbjected to sal ts for 4 successi ve. \\leeks .
~l)()
C\.' 1'1.-' ~l n ~Il'
r.... n~l1l{R;~I1~""J
Preparen!p~ril)d
(d~I.\~ )\:l=-".S I)(Ran~l')
46,6 ±. 2.0
( ,, (
Ink:\.>I ion
l).~ . .~
Sur\'iv~11
(i'; j""
:-.;IlL'tI
ding
Salt;..;
Conlrn]
.---------,r------.-~~-----.~--~~~---~_,_____~---_____tr--~-'--'-..-:Vl(';l:l I SL~·(h1!1~.:_' ~
! Ill'l ; j l'L r - 1 d I. 1J; l : i i ) ~ I
1~',!ll:I~'/Sj)~lil/ I rt!;I'...·.', \1-=-SI),l i ~ nu ];Ul\! I Ie] r] '~" I
n'~-I ~ II
1\1gS(}~ XO,() )7.4 ± 4.2 ~~
L')()·(-,O)
I () I I 1~L() ±
16, I :':;;: ::~
2~.() + 1.~.5
I39.7 +
( I 0--+ h)
~ 1()-...l J )
~). I ± l,).5 ;:~
( 15 -Hl)
1.+ ..'1 ±I 2'c) ;,~
( 1-77 )
.:47.7 ±~, I
!I
)(){)( 1
SOJHI
~(),( 1
~(),{)
7{). (\
KCI
CaS( ).j
ji
l. (), 1() I \
~--~--~---+~~~,----+--j-_._--~-~+-._------ ~-----~~~--~--l..+)l)() I ~4,(}:±:-+..1;:' 14." ~
I
Table 3: l~()tal pr()tcln content lllg/Illg tissue anel activIty ()f
ASAT & ALA l-'~ f\CP & AKl)unite/1l1g pr()tein in tiSSllCS of L.11 ([1({1e11 sis in (i i f fc re nt sa] ts fa r fc) ur sueccssive \veeks.
SaIl'..; pplll Prntci n ASAl ALAT }\C"P t\KP
Control 2! ,-+7 J.I ± 2.71 t Il,O ~ -+- (}.lJ I .'"' ±--
± 0.71 I,OY O.X2 {). ()( 12 o (J(lX
[\1 gS().l [~._~o 2X7 -+ 2.):') ± 1),027 + ().~()I ±(SO()) ± {), XS ~' 0.20 (), Jl) n. ()()(1 () ,()O:s_~ ;'- l
NaCL 20.~5 ~.os ± I.S5 ± l L(1) + l1,lJOlJ] !.-
L~{)() ) ± 4.()~ n.21 (). [lJ :( OJ)O.+ (},OOX :" ;::;;.
KCJ I X.t)h 2,99 ± 2.1 () ± (),()L~ ± O.O()6h ±(.~OO) ± l.l)) O.l.JO .0.20 () ,O(J2 :1'-;:- ().(}()! ):'-
CaS()~ 17-() ! .:"9 ± 2.06 ± O.{)()3 ± (J.OJ [ ±(.~()O} ± 3.~ :: OJ) ;~: O. I :) ().Oll~ ;':;~:; 0.020 ;,,:
Na~C()l 1.1, 2~ 1.50 + lJO ± O_O(l~ --.- 1(I.(lon +(Ion ) ± I .() I nAO :: () I () :'~ 0.\)( i3 .:: ,:, (} (l((l :' ,: 1
~----~
/.
V;
oV;'
I
"ij
»om
('-:r:?:""~r:"f.
........-'pc::,....'-":::s
~ 7' Z..... ..:;:;..
:;: 10
I"; -C' 'J, -+- '.,J-.)
~
'J . .":::)
-l".i-X~ '::;-.,
0"
'J.
0'.'~
.Q' ::;--.---J[0 ......J.C'- --1
1....J
'J,'JI>£
-.l --J C"£'+-:-0':.- ~. ~I...,) ~ ~
0. ~ lJ
?' .J 1 J- '.J-J I..J
':/..S('Jf
a· -..J%--..J
....'.j-J
'J)--.j
I.,)
'Jl
'J)
IJ
'l:.
'oj)
-+- 'JI
~;
' ..... .J
.4--
:z:[.J
-I-
~: ~~ 'J_' '~J [;!. ~ ~ ~ ,~ ~ !-,~ i:; 1" ~ ~
::~~
:-~,
,~'
i;~
'~Nf":~... ::~.: .. ~~~~. >~~~y.::~~ .. ~.
~.~; .:: .~-)~: .::: .....;:.:.),::~~:: ."
,". ~~ '~.' .
;:...
. <1",f$W';;
.~;.·.'ontt:· .'::tftrj;~ .." ~~:: ~ ~ .
~$j;
:Ai~IWi'.,~,
~~:~.····i~
l\.1arker C'nntro! KCI C-'aS{)~
Fi~. 1: SI)S-P;--\(,E pattern of l issue ~()l uhIe protei ns of Lynlllllco IIU t({ /CIlsi.\
exposed tu suhlcthal cone. of salts and controls, Polassiunl chloride. T11agnc
SiU111 sulphatc~ CttlciUlll sulphate. sodiuln 'carhonate and ~()diun1 chloride,
stuclicd by Nczlin (1997) \vho state(i that ablati()11 of osphraciill1l1(a chenlosensory organ) resulted in a significant increase or egglaying~ activity l)f L. S!OlfJl1l1Iis. No (10ul)t~ Osphradia are ChCl11()
recept()rs (Hynlan, ]967).l"he results (Tab. 2) indicated that the sllrvival rate at first
shedding of L. 1]Cl!(llensi,,,' expc)sed to F. g(f{{ll1tic(1 lniracidia andnlaintained at salts cone. V\las less than controls. The rate \vas 7()-8()o/c C()lll]Jared to 93.3 ck fc)r controls. The cone. of the usc<.isal ts did not ,)revent L, nt/to lellsis infection \\1 ith F. (1; i(~(ln tic({
kel)l at 50 ppnl Mg.So4 (31.25(10) with significant variation (P<0.01) fr()lll contr()ls (64.28Ckf). This reSlllt agreed \vith 1\1ostafa( 1997) \vh() fOllnd that n1aintenance of B. (lle.r(lne/rin{! at MgS()4~ NaCl, KC)~ CaSo4 & MnSo4 post exposure to ~S. 171(/1l.\'()lli
111iracidia significantly redllced th~. infection rates. Styznka-Jurewicz (1965) gave the saIne conclusion \vith F. hC/J(lticll. J\lso~
data (Tab. 2) sh()wed that the prepatent period of snai)s J11ain~
tained at MgSo4 was significantly longer than in c()ntro's~ 57.4& 46.6 days~ reslJectively. This was parallel to the results of Darel (II. (20()S) on Ro{/i.r nat('l/eTlsis and G{IIIJ({ fr{111C(11U/(I. the
552
vectors of F. ,f...~ig(/ntic(l. vv'hich prepatent periuds \vere 62.7 ex47. 5 ciays~ res])ecti ve Iy. The dala a1 so sho\ved l hat the total nu 111
ber of 111etacercariae/snail fronl infected sllail~ nlaintained atsalts V\/as significantly lo\vcr than 111 controls (p<O.OS).
The present study re\!ealed 1h~lt i11 reeled snail S ll1ai Iltal ned inMgS()~~ CaSo4 & Na.2Co; caused high .~uppre~sion in cercarial
productic)ll conlpared to controls. In ('aSo~ group. the nlt'~ln
nU111ber of llletacercariae \V(lS ] 4.5 COJl1llared to 59.3 rnctacer
cariae for control ones. This result a[!rees v/ith those of I\1ostafa0....-"
(1997) on schistosolne infected B. (lle.xLlll(!ri}}{J nlaintained at thesalts. The duration of shedding vVUS shortened for snails exposedto these salts cOll1pared to control. This period ranged fr()J11 25.6to 35. I days for snai Is Inaintai ned at the tested sal ts C01l1])arcc! to43.7 days f()r controls with significant variations (p«).Ol). l~he
present decrease in cercarial production under the salts f11ay bepartially due tt) short period of cercarial shedding.
No doubt~ these salts deteriorated the physiological activitiesof the snails (Tab. 3) as lnaintenance of L. llalo!ensis in Na~Co3
(100 ppln), CaSo4 (500 ppnl) and MgSo4 (SOC) ppnl) for foursllccessive weeks reduced the total protein value and activitiesof alllinotransferase and phosphatase enzynles in snail tissueshotnogenate conlpared to controls. The values were 13.23 nlg/g~
17.6 mg/g and 18.30 lng/g and the reduction rate were 38.49(J~
18% and 14.80/0 with significant difference (p«),05). Ishak et al.(] 975) and Bakry et ol (2003) noticed a decrease in the lllctab<)lie activities of trematode infected B. aJc);al1llrinti & B. frUllCO[
US~ a significant reduction in t()tal protein content of infected L.11(lt(llensis, respectively. This agreed with Gies (1988) \"/ho stat
ed that the hosts must have high levels of energy generatingnletabohtes to sustain nOfIna} nletabolic activities during the parasite development. Disturbing the snails ' energy nlet.abo)is]n bylTIolluscicides helped in rendering their physiologjcal processesless suitable for parasite developnlent. Also~ Bakry (2004) reported a large decline in ATP and total adenine nucleotide." le~
vels in tissues of B. a!exo!"1clrillo treated "lith JllethanoJ extract ofplant~ Furceraea g(l?antean and L{1111/Jranthus s/Jec(oIJilis.
The anl1notransferase enzylnes represent an inlportant linkbetween carbohydrate & anlino acid Inetabolic path~lays. Theactivities in tissue hOlllOgenate or blood of an Or2:anlSll1 \\Jerew ~
553
g()0 LJ t ()()1s ()f (1 etee t i() J1 () f \ J ~lr j at i()n in phY'sicd ()giC(J] J) r(1ce~s {, dtIle ()rg(tnislll (To)htJ ef (,r, 1997). 1~11e l)resent resLIJts revet-decl (.1
reCll1ctj(JIl in t}lC acti\lities ()f i\~S/\T~ /\I AT' & /\(~~P in tis~lle~ (Jf
." n~lj IS J11~li 11 taincc.i i 11 s~ll ts. "T"lle prc) n()ll fl ce(J reel llC t i()n i 11 t11e C Jl z~'
~,Illle.s cv:ti\'ities \'!a~ j II Sl1(111s fllainl,11rleCl III \J a,~(~() ~ {I (){)J l lliT1L
r-IIJle \',lllles f()r A·SAl~~ AIJi\l-' (~ f\CP \-\:ere 1.5 tl/l11g'. I .~~ l}/llj~
& ().()()4 L.l/l11g r)r()leiJl~ respectively' C()J11Ilarecl 1(' J. i U/!11g~ :.:
ll/lllg &, () .() ~~ il / I'llg 1) rt) te i t1 f(J r C(J 11 t r() I~ (};<() 0 () ~~ ) • rr'hc 1) ~ \ ~ S C" lit
rCl1ll\:tic)n ()r elev~lti()n in ASAT & l~I-Ji\ 1" acli\'jtie~ Jll(l)/ 11c Clue
t(') ,lei \:erse efrects () f t 11e s ~tJ ts ()n t}1e tisSIIe cells L. 11 {I r(IIe,-,sis.I~~.l\vj (J98f,) atl(l /\bci EI-l{clhecJl1 et (II. (1<)8(1) f()ll!1ci th;Jl (icc~
rCL.lSe 111 tr~lnsalllil1~lse ~tctivilies t()Ltlcl he allril)llleci l() hel,at()l()xic ]J()tency' tllr{)Llgh al)plic~ltjc)n ()f 11cstici{ies. Bl1t~ J--.I~l1l17a cl
(II. ( J 977) reI)()rtecl tllat tIle incre,lse in f\S/\ l' & ALAT ,-lel i vllics1111gJlt t,C tllle t() tt)Xi.lel111Ll. Batllr() ef (II. ( 19~)5) repl)rteci tll~}t C\-·
I){)Sllre IJ' /)(Ilifstr;s t() 11cXaclll()rc)l,eIlZCn stillllll~lled the, actl\·llj'
c)f 11()1)JS~lccll~lri(ies 11Y'droly/sing CllZ)-'llles atlcl de.teri{)r~.ltccl tileJllel,lb()]lS111 ()f rt~se,r,,!c glv'c()gcn \Vllich 111 hilliteci tIle 2.r()\vlh ~i11ll
~ ~ ~. _.incrcasecl sn~liJs· feCLlJl(iit)i A M()Jlte.il (lJ1(1 C'Jorci()J1 (1993) ll()ticcciall i 11 Crc ase ()r ;\CPill tisSLlC ()f i J1 fcc tec.i L. !r l.f Ii ( '( III {1(1 aIlli ~1t1r il")LIlCd it 1(1 the illcrc,lsc ()f }lv(il"())\',lSe e.nZ\!J11es. Hl)\\/e\lcr~ snai l~'
~ ~ ~
Ill'1il1tC.IlLll1CC ill Mg S()-l & CaS()~ f()r f()llr sLlcceSSlvc \\/ccks C~l-
llSCti ~l sigllificant e)c\/ati()1l In AKP aCllv'it)/ in C()1111)(lris())1 \\'~lt}'l
Cl)Jltr())s. Ttle actl\Jltic.s \vere ().2() 1 & ().().31 Ll/Illg pr()tcl11 resIJective))1 C()Jl1I}~lre(j t() G.() 13 Ll/ll1g l1rC)tein f()r C()lltr()}s (}J«).() I)
Tile clectr()IJl)()retic sel}~lr,lti()n ()f tisslle IlrC)tcll1S (f~ig. I. l-,~tl).
4 J frolll LJO Ilfl/(,lellSis sl'1()\ved clifferent effects ()f tIle testecl sa]t~
()Jl tl'1e S)!ntllc.sis ()f ]1r()tcin V/it}11Jl tIle snails )/lclliecl a C()Jl1plcx
J)~lttern ()f 11() IY"j)C IJl i(tes rallgi ng in lll()1cCLll ~lr \ve ight !lL~l \\'(~C r12~.()g ll) 2()S k l)a. TJlerc \vere three IJ~lll(ls Ilrescnt 111 c{)l1lr()l.~
an (! (I1s~111rC arcll fr() III treateli ()nes. "fhe Jll() lee u] ~lr \VL ig11 t~ () r t 11l~
h~lI1(tS \,verc 47.()~ 61.37 & 85.()8 k l),l. On Ihl~ (,)tllcr jl~Lnci, th(~re
\\Jere se\"cral 11UllCis ill the treatcej snails al1(I 11()t f{)LIIlCl 1n Cl)tl
tr()ls. S()ll1C of t}lC t1(\llcts \\:erc C()111111()Il1)' IJrcsent 111 s11Lll1s lrc<l
leet \vitl) KCl~ rv1~.S(t.i & (~~lS()_t \\'itl1 ~l r~ln~e (1r t11()lL'clll~lr \\-'Cl-L . L
UJll l1Cl\.\;ccn --+ 1.2 (l11(1 45 k Da. I-1()\\:e\'cr .. thl' ()llly l'()nll11{)11 lll.tllliL _
}) (' t \ \/e l-" 11 C() nt r\)]~ a11 It tre ate(t sn~ l i ]s \\/ a~ t f1~ It () f 7() .'+2 r() ~().):::; kI)~l. It \\'~l" \\'()J·thy IrteJlti()nin~ tllttl ()IlC !1anll \\-'~L~ llell~clt"d fClr- ~.
~n~Li)~ trC;tlL~(1 \\'ilh each ()r the tc~teLI s~_111~ antt neither (()Ullli 111
controls nor in other treated snails. The Il1o]ecular \vei£hts of theL
,banels \vere 28.68 k Da at NaCL 3().08 K Da at. J\1g.So~~ 53.84 kDa, 88.58 k Da & 1()().75 k Da at CaS()~ & 2()S k Da at KC~l.
The results also indicated djfferences in the !Jercent ()f C0J11rnOn
bands betvv!een contrc)ls and tested snails. F())" exallll11e. the handof 38.86 k D ash0 \.ve(1 11.2 o/(~ fareont1'01~ and 6 (}(~ fo r S 11 all S f X
}Josed t() CaSo4. that of 115.49 k Da sh()'v~recl 5,5 IS/(- for c'()n lr()L')
ZUlli 109(, fa r th()seat 1\1 a(~ 1. T11 e h1ghest rec()rded I!eree ill fu rbanets vv'as 16.6(k) & 15t;;() for those of 31.08 & 102.69 k l)iJ fr()1l1
snails exposed to CaSo~ ane! NaCl, resI)ectiveJy. This 111ay inclicale that SOllle factors increased or decreased dependj ng on thesal t llsed. Thi s alteration in protein fractlonatic)rl (Jnci I11etabc)] icl)fl)CeSSes cOllld be dlle to a production of additional IJrotcins ordinlinished ones by treated snails in addition tC) disturbance ()f
11()IYIJeIJtide lnetabolis111. El-Ansary et al. (20()O) used the sanletechniclue t{J separate tissue IJroteins of control anci lrenlatodcinfected B. clle.xolldriTl{{, and found correlation het\\-'een separa~
t 1()1l pattern and the llletab())ic redirecti()ll of the snail hc)st by thedeve]ol)ed sporocysts. EI-Khayat (2004) reported that the 111axiJ11Ull1 of protein fractions nUlllher and the Vv'idest range of llloleclllar weight were in prel11ature anti adults in B. CI!(!.'({1I1(lrini[; Is.tr;I(IIJrclt(1 and a hybrid of both. Although control and experil11enttal L. notc/lensi.\' possessed siJnllar !Jrotein ban(is~ they differeci inthe content of tTIolecular react on snail ]Jroductivity and survivorship. The data sho\ved the productivity of snails Inaintainedat MgSo~ was slightly higher than in controls. Vies versa \vasrecorded for snails at CaSo4. Thjs 111ay be due to the detectecl11ands of CaSo4 & MgSo4 snails ancl absence in other snails orcontrol (Tab. 4). The bands were one of 30.08 k Da for snails at.MgSo4 , three of 53.84~ 88.58 & 100.75 k Da for CaSo4.
It is concluded that the salinity in certain snail habitats inEg)lpt (specially iTl drains and \vater courses in ne\vly reclaill1edareas) could reach to high levels \vhich did not prevent infectionof L. JZ{J!lllensis \vith F. giKQ11tica although it should suppressinfection rates and cercaria) production that interrupt the parasltelransll11ssion. Consequently, fllrlher investigatlons are ongoingt() is()l ate such protei 11 detected in trealnlenl tC) characterize andto evaluate their effect on snails' survival and fecunditv .
.;
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