Don 1987 Aquacultural-Engineering

7/28/2019 Don 1987 Aquacultural-Engineering

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Aq uacu l tura l E ng ineering 6 (1987) 69-7 4

Increased Tilapia Em bryo V iabili ty U sing U ltravioletIrradiation in a C losed R ecirculating Z uger-B ottleSystemJ . D o n , M . K o i l l e r , O . Y e h e s k e l , R . R . A v t a l i o n

Departm ent of L ife Sciences, Bar-Ilan Un iversity, Ram at-Gan, I srae la n d

K . E . N u s s b a u mDepartment of M icrobiology,College of Veterinary M edicine, Au burn University,Alabama, USA

A B S T R A C TT h e a d d i t i o n o f a n u l t ra v i o l e t l i q u i d s te r i li z e r ( U V L S ) t o a c l o s e d r e c i rc u -l a ti n g - w a t er Z u g e r - b o t t l e i n c u b a t o r c o n s i d e r a b ly e n h a n c e d t h e v ia b il it y o fa r t if ic i a ll y f e r t i l i z e d e g g s . T h e s a m e a m o u n t s o f e gg s w e r e i n c u b a t e d i n t w os e p a r a t e Z u g e r - b o t t l e s y s te m s , e q u i p p e d w i t h t h e r m o r e g u l a t i o n ( 2 6 C )a n d c h a r c o a l f il te r s a n d w i t h o r w i t h o u t U V L S . T h e d i ff e re n c e i n e ffi -c i e n cy b e t w e e n t h e se t w o s y s t e m s i n c re a s e d o v e r ti m e f o l l o w i n g t h e i re x t e n s i v e u s e d u r i n g s e v e n w e e k s . W h i l e t h e e f fi c ie n c y o f t h e s e t w o s y s t e m sw a s q u i t e s i m i l a r f o r e gg s w h o s e i n c u b a t i o n s ta r ts u p t o t h e e n d o f t h e f ir s tw e e k ( 88 % ), t h e v i a b i l it y o f t h e e g gs i n c u b a t e d a t th e e n d o f t h e f i f th w e e k( fo r a n a d d i t i o n a l t w o w e e k s ) w a s h i g h l y a f fe c te d , m a i n l y i n t h e n o n -U V L S - e q u i p p e d s y s t e m ( 1 3 " 5 % ) i n c o m p a r i s o n t o t h e U V L S - e q u i p p e di n c u b a t o r ( 60 % ). T h i s d e c r e a s e o f v ia b i li ty s e e m s t o b e d u e b o t h t o a b r u p te x h a u s t io n o f c h a r c o a l a ct iv i ty a n d t o s l o w d e c re a s e o f U V L S e ff ic i en c y a sa r e s u lt o f s li m e a n d m i n e r a l d e p o s it s.

I N T R O D U C T I O NU l t r a v i o l e t r a d i a t i o n ( U V ) w a s f o u n d t o b e a n e f f e c t i v e a g e n t f o r n o n -c h e m i c a l d i s i n f e c t i o n o f w a t e r f o r a q u a c u l t u r e ( S p o t t e, 1 9 7 9 ; G i l p in e ta l . , 1 9 8 5 ). S a l m o n i d s i n U V - t r e a t e d w a t e r h a d a s i g n if ic a n t ly d e c r e a s e d

69Aquacul tural Engineer ing 014 4-86 09/8 7/S0 3.50-- Elsevier Applied SciencePublishers Ltd, En gland. 1987. Printed in G reat B ritain


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7 0 J . D o n , M . K o i l l e r , O . Y e h e s k e l , R . R . A v t a l i o n , K . E . N u s s b a u minc idence o f d i sease , and UV t rea tment was h igh ly l e tha l fo r bac te r i a ,v i ru s es , p ro t ozoa and o t he r mi c roo rgan i s ms (Bur rows and Combs ,1968 ) . In an aqua r i um o f s ea wa t e r , 99% o f s u s pended bac t e r i a wereinac tiva ted by a U V s te r il izer (Hera ld e t a l . , 1962) . The effect iveness ofU V for d i s in fec tion i s limi ted by the v ery sha llow pen et ra t ion o f U V in toturb id wa ter (C olbe rg an d Lingg, 1978 ; Phill ips, 1983).The t i l ap ia subgenera S a r o t h e r o d o n a n d . O r e o c h r o m i s (Trewavas ,1982) a re cur ren t ly used in our l abora to ry fo r the s tudy of po lyp lo idy ,gynogenes i s and t rans fer o f immuni ty f rom the mother to the embryo .Eggs are ar t i f ic ia l ly fer t i l ized, or o therwise manipulated, and thenincubated in a Zuger -bo t t l e sys tem, as p rev ious ly descr ibed (Don andAvta l ion , 1986; Rothbard and Hula ta , 1980) . A UV l iqu id s t e r i l i zer(UV LS) was tes ted in an a t t em pt to overco m e h igh embryonic mor ta li ty .

M A T E R IA L S A N D M E T H O D SI n c u b a t o rTwo separa te c losed rec i rcu la t ing incubator sys tems each o f 20 Zugerja rs were used s imul taneous ly . Both sys tems were equ ipped wi thac t iva ted charcoa l f i l t e r s and independen t t empera tu re-con t ro l l ed 750-l i ter reservoi rs . Flow to each Zuger jar could be regulated separately .The UVLS (TANA-10 , UV l iqu id s t e r i l i zer , NH Indus t r i es , Nt ivHalamed Hei , I s rae l ) was ins ta l l ed in the water feed l ine o f one o f thei ncuba t o r s . T he UVL S has t wo l ow p res s u re H g UV t ubes p roduc i ngapprox imate ly 3 x 104 k tW cm -z wi th 85% of rad ia t ion be ing 2 53 -7 nm.Maxi mum f l ow t h rough t he UVL S i s 30 l i t e r mi n - t . L amp ou t pu t i sm on i t o red by an am met e r .P r o c e d u r eBefo re expe r i men t s s t a r t ed , bo t h t he UVL S-equ i pped and t he non -UVLS-equipped incubators were c leaned thoroughly and d i s in fec tedwith hypochlor i te . Both f i l ters were c leaned and repacked wi th f reshact ivated charcoal . The incubator was then f i l led wi th f resh tap water .Du r ing the spaw ning per iod (Ap r i l -A ugus t ) , a se r ies o f b roo ds o f ar ti fi -cially fertil ized eggs of O. a u r e u s a n d O . n i l o t i c u s were s imul taneouslyand equal ly d is t r ibuted in to the jars in both incubators , as previouslyrepor ted (Don and Avta l ion , 1986) . The incubators were ex tens ive lyused dur ing th ree per iods o f exper imenta t ion o f seven weeks each .Before each per iod o f exper imenta t ion , the sys tems were thoroughly


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Increased tilapia em bryo viability using ultraviolet irradiation 71c l e a n e d , a s d e t a i l e d a b o v e . E a c h w e e k , g r o u p s o f 1 5 0 t o 2 0 0 f e r t i l i z e de ggs w e re d i s t r i bu t e d e qua l l y i n dup l i c a t e o r t r i p l i c a t e i n t o t he j a r s o fb o t h s y st em s . T h e n u m b e r o f t h e e m b r y o s w a s c o u n t e d a t fo u r d i ff e r en te m bry on i c st age s : m oru l a (M ) , 6 - 8 h a f t e r f e r ti li z a t ion ; p i g m e n t a t i on (P ),4 5 -6 0 h a f t e r f e r ti l iz a t i on ; ha t c h i ng (H ), 7 2 -8 0 h a f t e r f e r t il i z a ti on ; a n dv i a b il it y ( V ) b e y o n d t h e y o l k s a c a d s o r p t i o n a t 1 2 - 1 3 d a y s a f t e r f e rt il iz a -t io n . T h e r e s u lt s o b t a i n e d w e r e a n a l y z e d s t a ti st ic a ll y u s i n g th e S t u d e n t t-t es t. L i v i n g b a c t e r i a i n w a t e r o f b o t h j a r s a n d r e s e r v o i rs w e r e c o u n t e d b yp l a ti n g in b r a i n - h a r d a g a r p l a t es .

R E S U L T S A N D D I S C U S S IO NT h e r e s u lt s p r e s e n t e d i n F ig s 1 a n d 2 s h o w t h a t t h e u se o f U V L S w a sc l e a r l y be ne f i c i a l t o t he f i na l v ia b i l it y o f t i l a p i a e mb ryo s ( e .g ., 60 + 10 .6%w i t h U V L S i n c o m p a r i s o n t o 1 3 "5 _+ 1 2 % w i t h o u t U V L S a t t h e e n d o f as e v e n - w e e k u s e o f w a t e r i n a c l o se d s y s te m ) . T h e d i f f e r e n c e b e t w e e n t h et w o s ys t em s w a s s t il l s ma l l up t o t he s e c o nd w e e k a f t e r c l e a n i ng , bu t i ts t a r t e d t o b e s t a ti s ti c a ll y s i gn i fi c a n t a t t he t h i rd w e e k , m a i n l y fo r H a n d V

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0 IM P H . V M P H V UV -U V

Fig. 1. Percentage of survivals (+S.D .) at the f if th week after cleaning for U V LS -equ ipped (+ U .V .) and non-U VL S-equ ipped ( -U .V . ) incuba to r s a t d if fe rent deve lop-mental stages of the e m bry os. M, morula; P, pigmentation; H , hatching; V, viabilitybey ond yolk sac adsorpt ion .


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72 J . D o n , M . K o i l l e r , O . Y e h e s k e l , R . R . A v t a l i o n , K . E . N u s s b a u m

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~_ 40Zwz c

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Fig . 2 . Tim e decline in the percentage of em bryo viability ( _+S.D.) at the V stage. A ,+ U.V.; m, - U.V.

s t a g e s ( P < 0 . 0 2 5 a n d 0 " 0 1 , r e s p e c t i v e l y ) ( T a b l e 1 ) . I n t h e e x p e r i m e n tw h i c h s t a r t e d a t t h e f i f th w e e k f o l lo w i n g th e c l e a n i n g o f t h e s y s t e m , a lls t a g e s , e x c e p t M , w e r e a f f e c t e d i n t h e n o n - U V L S - e q u i p p e d s y s t e m .C o n s e q u e n t l y t h e d i f f e re n c e s i n v i a b il it y b e t w e e n t h e t w o i n c u b a t o r sw e re h igh ly s i gn i fi c a n t : P < 0" 005 fo r s t a ge s P , H a nd V (T a b l e 1 a nd F ig .1 ) . A t no t im e d id t he M-s t a ge e ggs e x h ib i t a ny d i f f e r e nc e i n v i a b i l i t y .T h e d e c l i n e i n v i a b il it y o f e m b r y o a t th e V s ta g e s e e m s t o b e l i n e a r w i t ht im e (F ig . 2 ). T h i s s ugge s t s a c on s t a n t de c l i ne i n w a t e r qua l i t y , p ro ba b lydue t o t he a c c um ula t e d e f f e c t o f t ox i c a n t s t ha t a c c rue a s a r e s u l t o f t hec h a n g e s i n U V L S e ff ic ie n cy . T h i s m i g h t b e p r o d u c e d b y a d e c r e a s e dc a p a c i t y o f t h e U V l a m p d u e t o s li m e a n d m i n e r a l d e p o s i ts , a s w e l l a s t othe e x ha us t i on o f t he c ha rc oa l f i l te r ac t iv i ty . I n f a c t, a s u rve y o f t he w a t e rb a c t e ri a l c o n t a m i n a t i o n s h o w e d a c o n s t a n t h i g h n u m b e r o f l iv in gb a c t e r i a i n t h e Z u g e r - j a r w a t e r a n d i n t h e r e s e r v o i r o f t h e n o n - U V L S -e q u i p p e d s y s t e m (T a b l e 2).

A C K N O W L E D G E M E N TT h i s r e s e a r ch w a s s u p p o r t e d b y a g r a n t ( 1 - 7 1 5 - 8 3 ) f r o m t h e U n i t e dS t a t e s - I s r a e l B i n a t i o n a l A g r i c u l t u r a l R e s e a r c h a n d D e v e l o p m e n t F u n d( B A R D ) .


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I nc r eased i lap ia em bryo v iab i l i t y us ing u l t rav io le t r rad ia t ion 73

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74 J. Do n, M. Koiller, O. Yeheskel, R. R. Av talion , K. E . Nussb aumR E F E R E N C E S

Bur rows , R . E . & Com bs , B . D. (1968) . Con t ro l l ed env i ronmen t s f o r s a lmonpropagat ion . Prog. Fish Cu l t. , 3 0 , 1 2 3 - 3 6 .Colberg , E J . & Lingg, A. J . (1978) . Ef fec tor of ozonat ion on microbia l f i shpa thogen s , am m onia , n i t ra te and B O D in s im ula ted reuse ha tchery water. J .Fish Res . Bd. , Canada, 35, 10 .D on , J . & A vta lion , R. R. (1986) . Indu ct ion of t rip lo idy in Oreochromis aureusby h ea t -shock . T h e o r Ap p l . G e n e t., 7 2 , 1 8 6 - 9 2 .G i lpin, R. W ., D i l lon, S. B. , K eyser , P., A nd rok i tes , A. , B erub e, M ., Carpe nd ale ,N. , Skoring, J ., Hurley, J . & K aplan, A . M . (1985 ) . Dis infec t ion of c i rculatingw ater sys tems by u l t rav io le t l igh t and ha logenat ion , W ater Res ., 1 9 , 8 3 9 - 4 8 .H era ld , E . S ., D em pster , R. P ., N ol te rs , C. an d Hu nt , M . L. (1962). F i l tra t ion andul t rav io le t st e r il iza t ion of sea w ater in l a rge c losed a nd sem i-c losed aquar iumsystems. Bull. Inst . Oceanogr. , 1 B , 4 9 - 6 1 .Phillips, R. (1983). Sources a nd A pp l ica t ions o f Ul t rav io le t Rad ia t ion , A c a d e m i cPres s , Lo nd on , pp . 5 -11 .R othba rd , S . and Hula ta , G . (1980) . Close d sys tem incubator for c ich l id eggs .Prog. Fish C ult . , 4 2 , 2 0 3 - 4 .Spot te , S. (1979) . Fish and Inver tebra te Cu l ture -Water Managemen t in ClosedSystems, Joh n W i ley , N ew York , p . 145 .Trewavas, E. (1982) . Ti lapia: taxonomy and speciat ion. In: The Bio logy andCul ture o f T i lap ias , ed . R . S . V . Pu l li n and R . H . Low e-McC onne l l , ICL A R Medi t ion , pp . 3- 13 .

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Don 1987 Aquacultural-Engineering