Leger 1982 Aquacultural-Engineering

7/28/2019 Leger 1982 Aquacultural-Engineering

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Aquacultural Engineering 1 (1982) 45-53

A U T O M A T I O N I N S T O C K - C U L T U R E M A I N T E N A N C E A N DJ U V E N I L E S E P A R A T I O N O F T H E M Y S I D

M Y S I D O P S I S B A H I A ( M O L E N O C K )

PHILIPPE LINGER and PATRICK SORGELOOS~:Artemia Reference Center, State University of Ghent, J. Plateaustraat 22, B-9000 Ghent, Belgium

ABSTRACTA ba t ch s y s t em wi t h subgrave l f i l t e r f o r s t ock - cu l t u re m a i n t enance o f M y s id o p s is b a h i ain ar t if i cia l sea wa ter i s descr ibed . The feedin g o f my s ids o n A r t e m i a i s op t i m i zedthrough the cyc l ic pum pin g o f br ine shr imp ins tar I naup l i i f ro m a ref rigerator to thes tock-cul tures . A n easi ly c ons t ruc ted incubator-separator apparatus is descr ibed fo rthe s tanda rdized harves t ing o f m ys id ]uveni les to be u sed as t es t organisms .

INTRODUCTIONT h e e s t u a r i n e m y s i d s h r i m p M ys i dops i s bah i a M olenock (1 969 ) i s r egu l a r ly be ing u sedas a su i t ab l e t e s t o rgan i sm in t ox i co log i ca l (Bahne r et al . , 1 9 7 7 ; N i m m o et al . , 1977 ;A n o n y m o u s , 1 9 7 8 ) a n d n u t r i t i o n a l s t u d i e s ( J o h n s et al. , 1981) . The re fo r e M ys i dops i sbahia j uven i l e s we re s e l ec ted a s t e s t o rgan i sms i n s t anda rd i zed p r ed a to r -p r ey expe r i -m en t s t o eva lua t e the nu t r i t i ona l va lue o f va r i ous A r t e m i a produc t s , e . g . decapsu l a t edcys t s and naup l i i f r om d i f f e r en t geog raph i ca l o r i g in , va r i ous p r ep a ra t i ons o f f ed andunfed naup t i i , e t c .

Acc o rd ing t o t he l i t e r a tu r e s t ock cu l t u r e s o f t h is mys id shou ld be m a in t a ined i no p e n f l o w - t h r o u g h s y s t e m s w i t h n a t u r a l s e a w a t e r ( C l u t t e r a n d T h e i l a ck e r , 1 9 7 1 ;A n o n y m o u s , 1 9 7 8 ). A n o r i g in a l t e c h n i q u e w a s w o r k e d o u t f o r s t o c k m a i n t e n a n c e i nba t c h sys t em s w i th a r t if i c ia l s ea wa t e r i n s it ua t i ons whe re no runn ing s ea wa t e r i sa v a il ab l e. F o o d d i s t ri b u t i o n t o t h e s t o c k c u l t u r e s w a s a u t o m a t e d a n d a n i n c u b a t o r -s e p a r a t o r s y s t e m d e v e l o p e d f o r t h e s t a n d a r d iz e d h a r v e s ti n g o f m y s i d j u v e ni le s t o b eu s e d a s t e s t o r g an i s m s i n t h e p r e d a t o r - p r e y e x p e r i m e n t s .t Research assistant and ~ research associate at the National Fund for Scientific Research (Belgium).

45Aquacultural Engineering 0144-8609[82[0001-0045]$02.75 Applied Science Publishers Ltd,England, 1982Printed in Great Britain


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4 6 P . L I N G E R, P . S O R G E L O O Sa i r 4 ~ a ir

2

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F i g . 1 . S c h e m a t i c d i ag ra m o f t h e a q u a r i u m s e t - u p f o r s t o c k - c u l t u r e m a i n t e n a n c e o f Mysidopsisbahia w i t h d e t a i l s o n t h e s u b g r a v el f i lt e r s y s t e m . ( 1 ) I n f l o w o f f o o d , ( 2 ) a i r -w a t e r l i ft , ( 3 ) su b -g r a v e l f i lt e r s y s t e m , ( 4 ) m a r i n e s a n d , ( 5 ) c r u s h e d s e a s h e l l s , ( 6 ) s i l e x g r a i n s , ( 7 ) p e r f o r a t e d p l a s t i cp l a t e .

S T O C K - C U L T U R E M A I N T E N A N C E

G l a ss a q u a r ia ( 6 0 3 0 3 5 c m ; c a p a c i t y 6 0 l it r e s) w e r e e q u i p p e d w i t h a s u b g r a v e lf i lt e r a s s c h e m a t i c a l l y o u t l i n e d i n F i g . 1 . A p e r f o r a t e d p l a s t i c p l a t e w a s in s t a l l e d a t t h eb o t t o m o f e a c h a q u a r i u m a n d c o v e r e d w i t h t h r e e f i l t e r i n g l a y e r s : i . e . s i l e x - g r a i n s a t t h eb o t t o m ( a v e ra g e p a r t ic l e s i z e 3 r a m ) , f i n e m a r i n e s a n d a t t h e t o p , a n d a t h i n l a y e r o fc r u s h e d s e a s h e ll s in b e t w e e n . T w o a i r- w a t er li f ts p e r a q u a r i u m c i r c u l a t e d t h e w a t e rt h r o u g h t h e s u b g r a v e l f i lt e r a t a f l o w r a t e o f 5 0 - 8 0 l it r e s r ai n 1 m - 2 f i lt e r b e d , a s s u r in go p t i m a l b a c t e r i a l a c t i v it y i n t h e f i lt e r ( K i n n e , 1 9 7 6 ) .

A r t i f ic i a l s e a w a t e r p r e p a r e d a c c o r d i n g t o t h e f o r m u l a o f D i e t r i c h a n d K a l le (K a l l e ,1 9 7 1 ) b u t e n r i c h e d w i t h N a H C O 3 u n t i l t h e p H s t a b il iz e d in t h e r a ng e 8 . 0 - 8 . 5 ( S p o t t e ,1 9 7 9 ) , w a s s e l e c t e d a s a s u i ta b l e m e d i u m . T h e s a l i n it y o f th e 0 . 2 g m f i lt e r e d s e a w a t e r


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AUTOMATIONOF MYSID STOCK-CULTUREMAINTENANCE 47was adjusted to 30 ppt and the temperature maintained at 23C -+ 2C (Johns, personalcommunication).

Since mysids prefer dimmed light conditions (Kinne, 1977) the top and front sideof the aquaria were covered in such a way as to allow only diffuse indirect illumina-tion at 10-50 lux at the back side of the aquaria. The fluorescent light tubes in theculture room were set at a 12 h light, 12 h dark photoperiod.

In view of their high food value for M y s i d o p s i s b a h i a (Johns e t a l . , 1981)and theirlow content of chlorinated hydrocarbons (Olney et a l . , 1980), A r t e m i a nauplii fromthe Macau-Brazil variety were selected as food source. One food distribution per dayas advised in the literature (Clutter and Theilacker, 1971) resulted in an inefficientfood uptake, i.e. satiated juveniles and adult mysids continued to chase brine shrimpnauplii and bite them to pieces without consuming them. Whereas these decayingA r t e m i a fragments were continuously washed out in open flow-through systems, theyoverloaded the subgravel filter in batch systems. Maximal food uptake by the mysidswith minimal pollution effects was assured by four distributions per day of about40000 A r t e m i a nauplii to each aquarium (see below for technical details on auto-matic food-distribution).

The mysid stock cultures have been successfully maintained in batch condit ions forover one year. Each stock-aquarium continuously held a few hundred gravid females.Aside from the work concerning food preparation and distribution (see below) thefollowing operations were performed routinely:

1. Daily from Monday to Friday the filter bed was raked and quick checks weremade on water temperature, air-water lift activity, food presence, and overallstatus of the population (abnormal mortality, behaviour of adults);

2. Although in previous experiments the water quality was never a problem, thedissolved oxygen content, salinity, ammonia-N, nitrite and nitrate level weremonitored once a week. As soon as the nitrate level exceeded 20 ppm, 50% ofthe culture water was changed;

3. Every month the reproductive activity of the population was checked, i.e. a fewspawnings were controlled for the size of the brood, the viability, and malforma-tions of the juveniles. As a function o f age, a healthy female was carrying from 5to 20 young;4. As advised by Kinne (1976), the filter bed was renewed at least once a year.

A U T O M A T I O N I N F E E D I N G O P E R A T I O N S

In order to minimize the manual work of feeding the three 60 litre culture-stocks withA r t e m i a nauplii, the A r t e m i a cyst incubation and nauplii distribution was automatedthrough the operation of a microprocessor universal timer (TMS 1122, PBJ Electronik,Denmark). Manual intervention was required only three times a week for about 15 min.


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4 8 P . L E G E R , P . S O R G E L O O S

C 2 ] : [ l - - - 3 ( 3 1 D " :

A g ~ j j ~

CFig. 2. Set-up for the au tom atic incu bat io n and hatching of Artemia cysts. A, plastic bag w ithd r y c y s t s; B , p l a s t ic b a g w i t h i n c u b a t e d c y s t s ; C , s e a w a t e r r e s e r v o ir ; ( 1 ) c o n t i n u o u s a e r a t i o n ;( 2 ) f l u o r e s c e n t l ig h t t u b e ; ( 3 ) a e r a t i o n f r o m a i r -p u m p a c t i v a t ed b y t i m e r .

T h e s e t u p f o r n a u p l i i p r o d u c t i o n is s c h e m a t i c a l l y o u t l i n e d i n F i g . 2 . T h r e e t i m e s aw e e k 2 5 g M a c a u c y s t s w e r e h a t c h e d i n 1 0 l i tr e s o f s e a w a t e r a t 3 0 p p t s a l i ni ty , 2 3 C ,a n d u n d e r c o n t i n u o u s i l l u m i n a t i o n ( S o r g e l o o s , 1 9 8 0 ) . I n o r d e r t o a ss u re a m a x i m u mh a t c h i n g e f f i c i e n c y a s w e ll a s a h o m o g e n e o u s h a r v e s t o f i ns t a r I n a u p l i i, t h e c y s t s w e reh y d r a t e d a n d i n c u b a t e d e x a c t l y 3 0 h p r i o r t o h a r v e s t i n g , i .e . a t t h e p r es e t m o m e n t ,a i r - p u m p ( 3 ) w a s s w i t c h e d o n b y t h e t i m e r a n d s e a w a t e r w a s p u m p e d f r o m r e s er v o ir( C ) i n t o t h e h a t c h i n g b a g c o n t a i n i n g t h e d r y c y s t s .

M a n u a l h a r v e s t i n g o f th e A r t e m i a n a u p l i i w a s d o n e a s f o l l o w s : t h e a e r a t i o n w a ss w i t c h e d o f f a n d a b l a c k p l a s ti c c y l i n d e r w a s f i t t e d o v e r t h e h a t c h i n g b a g , l e av i ng t h el o w e r p a r t o f t h e f u n n e l i l lu m i n a t e d . D u e t o t h e i r lo w s p e c if i c g r a v it y , t h e e m p t y c y s ts h el ls f l o a t e d a t t h e w a t e r s u r fa c e w h i le t h e p h o t o t a c t i c n a u p l i i o f M a c a u -o r i g in c o n -c e n t r a t e d i n t h e f u n n e l o f t h e h a t c h i n g b a g . A f t e r 1 0 r a i n s e p a r a t i o n t h e n a u p l ii w e r es i p h o n e d o f f o n a 1 1 0 / a m f i l t er s c r e en , r i ns e d w i t h c h i l l e d s ea w a t e r ( 3 0 p p t , 2 - 5 C )a n d d i l u t e d t o a f i n a l c o n c e n t r a t i o n o f a b o u t 4 0 0 0 n a u p l ii m 1 -1 i n t o t h e f o o d s t o c kc o n t a in e r ( 1 l i tr e c a p a c i t y , s ee F ig . 3 ) wh ic h w a s i n s t a l le d i n a r e f r i g e r a to r . A u t o m a t i o nin f o o d d i s t r i b u t i o n wa s g r e a t l y f a c i l i t a t e d b y t h e p o s s ib i l i t y o f s t o r i n g l iv e i n s t a r IA r t e m i a i n t h e r e f r ig e r a t o r , i. e. p r o v i d e d t h e n a u p l i i w e r e c o n t i n u o u s l y k e p t i n s u s p e n -s io n b y s l ig h t a e r a t i o n , o v e r 9 5 % s u r v iva l w i th m in im a l l o s se s o f t h e i r i n d iv id u a l d r yw e i g h t w a s o b t a i n e d a f t e r 4 8 h s to r a g e o f M a c a u - n a u p l i i a t 2 - 5 C ( I A g e r e t a l . , 1981 a ) .

F o u r t im e s a d a y t h e t im e r a c t i v a t e d t h r e e e a s i l y a s s e m b le d p e r i s t a l t i c p u m p s ( s e eF ig . 4 ) a s s u r in g t h e d i s t r i b u t i o n o f n a u p l i i f r o m th e s t o c k - c y l i n d e r i n t h e r e f r i g e r a to rt o t h e t h r e e s t o c k - a q u a r ia . A f e w s e c o n d s b e f o r e t h e p u m p i n g s t o p p e d , t h e f e e d l in e sw e r e a u t o m a t i c a l l y r i n s e d w i t h s e a w a t e r , i. e. t h e e l e c t r o m a g n e t i c v a lv e ( 1 ) o p e n e d a n ds e a w a t e r w a s d r a i n e d i n t o t h e f o o d d i s t r i b u t i o n t u b e ; as a r e s u lt o f th e h y d r o s t a t i c


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A U T O M A T I O N O F M Y S I D S T O C K - C U L T U R E M A I N T E N A N C E 4 9(2 )

(3 )

F i g . 3 . S c h e m a t i c d i a g r a m o f t h e f o o d s t o c k - c o n t a i n e r w i t h a c c e s s or i e s. ( 1 ) E l e c t r o m a g n e t i cv a lv e a c t i v a t e d b y t i m e r , ( 2 ) i n f l o w o f s e a w a t e r f r o m r i n s i ng r e s e rv o i r , ( 3 ) f e e d l i ne c o n n e c t e dw i t h i n f l o w t u b e o f p e r i s t a l ti c p u m p s , ( 4 ) o n e - w a y v a l ve .

pr e ssu r e o f t he s ea w a t e r i n f low , t he one - w ay va lve ( 4 ) c l o sed o f f t he supp l y o f naup l ii ,and c l ean s ea w a t e r w as pum ped t h r oug h t he f eed li ne s i n t o t he s t ock - aqua ri a .

The p r e sen t au t o m a t i ons i n f oo d p r epa r a t i on and d i s t ri bu t i on no t on l y g r ea t lyf ac i l it a t ed bu t a t t he sam e t i m e op t i m i zed m ys i d s t ock - cu lt u r e m a i n t enanc e . Manua li n t e r ven t i ons on Mondays , Wednesdays , and F r i days w e r e l i m i t ed t o t he f o l l ow i ngope r a t i ons :

1 . i n t e r r u p t i on o f t he ae r a t i on i n ha t ch i ng bag A (o r B)2 . ha i ve s ti ng o f t he naup l i i f r om ha t ch i ng bag A ( o r B)3 . c l ean ing o f the f o od s t ock - con t a i ne r i n t he r e f r i ge r a t o r4 . t r ans f e r o f t he naup l ii in t o t he f o od s t ock - con t a i ne r5 . r insing of ha tchin g bag A (or B)6 . r e s t o ri ng t he ae r a t i on i n ha t ch i ng bag A ( o r B)7 . i npu t o f 25 g cys t s i n ha t ch i ng bag B (o r A )8 . f i ll ing of the sea water reservoi r w i th 10 l i tres of 3 0 p pt sea w ater9 . t r ans f e r o f t he t u be f o r s ea w a t e r i n f l ow t o ha t ch i ng bag B ( o r A )


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50 P . LEGER, P . SORGELO OS(1)

a - bFig. 4 . Schem at ic d iagram of per is ta l tic pum ps for Artemianaupl i i d is t r ibut ion. (1) Inf low (3 mminner d iam eter ) connec ted w i th fo od stock-container , (2) s i licone tubing (6 mm inne r d iameter )f ixed on tw o suppor ts , (3) ro ta t ing PVC-cam, (4) out f low (1 mm inner d iameter ) to s tock-aquar ium,(5) Crouz et synchronous m otor - red uctor 20 rpm, (6) e lect r ic act ivat ion by t ime r .

INCUBATOR-SEPARATOR FOR THE STANDARDIZED HARVESTING OFMYSID JUVENILES

F o r c o m p a r a t i v e n u t r i t i o n a l a n d t o x i c o l o g i c a l t e s ts w i t h m y s i d j u v e n i le s , i t is a l w a y si m p e r a t i v e t o u s e e x p e r i m e n t a l a n i m a l s o f a s p ec i fi c ag e . T h e c o n t i n u o u s c u l t u r es y s t e m f o r Artemia, Daphnia, a n d o t h e r i n v e r t e b r a t e s as d e v e l o p e d b y S o r g e l o o s a n dP e r s o o n e ( 1 9 7 3 ) w a s m o d i f i e d f o r s h o r t - te r m i n c u b a t i o n o f g ra v i d m y s i d s , a n d a u t o -m a t i c s e p a r a t i o n o f t h e i r 0 - 2 4 h o l d o f f sp r i n g . A d v a n t a g e w a s t a k e n o f t w o b e h a v i o u r a lc h a r a c t e r i s t i c s o f m y s i d s h r i m p s i n t h e d e v e l o p m e n t o f t h e p r e s e n t i n c u b a t o r - s e p a r a t o rs y s t e m :

a n i n c r ea s e o f t h e w a t e r - t e m p e r a t u r e b y 2 C s t i m u l a t e s t h e f e m a l e s t o re le a s et h e i r o f f s p r in g (J o h n s , p e r s o n a l c o m m u n i c a t i o n ) ;

- u p o n b i r t h j u v e n i l e m y s i d s d o n o t s w i m b u t s i n k t o t h e b o t t o m o r a r e c a r r i e da w a y b y c u r r e n t s . A f ew m i n u t e s l a t e r th e y m o u l t i n t o a s w i m m i n g s t ag e .

T h e a p p a r a t u s ( F i g . 5 ) w a s c o m p o s e d o f t w o i n t e r c o n n e c t e d c y l i n d r o c o n i c a l v e ss e ls ,w h i c h w e r e e n t i r e l y m a d e o f a c r y l ic g la ss a n d t h r o u g h w h i c h w a t e r c i r c u l a t e d b y t h ef o r c e o f a n a i r - w a t e r li f t . V e s s e l A ( 2 5 li t r e c a p a c i t y ) w a s e q u i p p e d w i t h a f i l te r c y l i n d e r


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AUTOMATIONOF MYSIDSTOCK-CULTUREMAINTENANCE 51~2J

~ - ( 3 )! , i t

\ \ /\ ' , /

~ ~x'4air

\\ /

71Fig. 5 . Schem at ic d iagram of incubator -separator apparatus for juveni le harvesting inM ysido psisbahia. A, incub ator for adul t mys ids wi th f i l ter baske t (1) , aerat ion line (2) , thermos tat -heater (3) ,and stopcock (5) w i th large opening; B, separator box for m ysid juveni les wi th f i l ter screen (4),watert ight f ixing system (a) of l id and stopcocks (6 and 7) .

( 1 2 0 0 / ~ m s c r e e n ) w h i c h r e t a i n e d a d u l t m y s i d s b u t l e t j u v e n i l e m y s i d s a n d b r i n e s h r im pn a u p l i i p a s s . W a t e r in v e s s e l A w a s a e r a t e d w i t h a n a i r s t o n e a n d k e p t a t 2 5 C w i t h as u b m e r s i b l e h e a t e r w i t h a t h e r m o s t a t . S t o p c o c k ( 5 ) a t t h e e n d o f t h e f u n n e l h a d aw i d e o p e n i n g ( 1 0 m m i n n e r d i a m e t e r ) t o f a c i l i ta t e pa s s ag e o f t h e m y s i d ju v e n i l e s .


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52 P. LEGER, P. SORG ELOOS

Vessel B (4 litre capacity) was made up of a cylindroconical bottom part on towhich a watertight cover was fixed with rubber bands and spring-back binders. Thebottom part held a 500/am filter screen, which for about half its surface was coveredby silicone strips that provided shelters for the mysid juveniles and at the same timeassured a more efficient suction of the A r t e m i a nauplii through the filter. The coverof vessel B was equipped with a purge tap (6) and with an open tube to be connectedto stopcock (5). The funnel was connected to the lateral air-water lift which pumpedwater and A r t e m i a nauplii from B into A.

A homogenous dispersion of the A r t e m i a nauplii over the entire apparatus wasfurthermore achieved by shading vessel A, i.e. the phototactic A r t e m i a nauplii (fromMacau origin) migrated to the unshaded bottom part from where they were easilycirculated through B and back into A.

A few hours prior to routine operation, the incubator-separator was filled with arti-ficial sea water (formula of Dietrich and Kalle, 30 ppt salinity, 0-2/lm filtered) and theheating and aeration were switched on. In order to attain a food density of about oneA r t e m i a nauplius per ml of culture medium about 30 000 freshly hatched nauplii wereadded to vessel A. As soon as the temperature had stabilized at 25C -+ 2C (warmerthan in the stock aquaria), as many gravid females as the number of juveniles wantedwere collected from the stock cultures and transferred into the filter basket o f vessel A.

After 24 h incubation a first batch of mysid juveniles could be harvested: i.e.stopcock (5) and the air were closed off while stopcock (6) was opened. Upon discon-necting the air-water lift tubing located just after stopcock (7), water could be drainedoff from B. As soon as the cover of vessel B was empty, stopcock (7) was closed, vesselB opened, and the mysid juveniles were collected.

If more juveniles were needed, vessel B was reassembled and connected to A, and anew batch of freshly hatched A r t e m i a nauplii was added. The second separationalways resulted in the greatest number of juveniles. More separations could be per-formed but the number of juveniles was sharply decreasing from the third separationonwards.

As soon as the separation was finished the adult mysids were collected from vesselA and put back into the stock-aquaria. The entire apparatus was rinsed with tap waterand air dried.

This incubator-separator system has been successfully used for routine productionof hundreds of mysid juveniles at a time. The high reproducibility of survival andgrowth results obtained with these juveniles in standard culture tests (IAger et al.,1981b) were the best proof of the suitability of this new incubator-separator systemfor the production of experimental test-organisms.

ACKNOWLEDGEMENTS

We are very indebted to Allan D. Beck and D. Michael Johns from the US-EPAEnvironmental Research Laboratory at Narragansett (RI-USA) for providing us with a


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A U T O M A T I O N O F M Y S ID S T O C K - C U L T U R E M A I N T E N A N C E 5 3

s t ock - cu l t u r e o f Mysidopsis bahia and f o r t he i r i n f o r m a t i on w i t h r ega rd to m ys i dcu l t u r ing ; t o Pau l V anhaecke f o r t he t r ansp or t o f t he m ys i ds t o ou r l abo r a t o r y ; t oFr ed dy D e t r y and Joh an Tave r n i e r f o r t he ir sugges ti ons and t e chn i ca l he l p in deve l op -i ng the va r i ous sys t em s , and t o E dm ond e J a spe rs f o r p r oo f r ead i ng the m anusc r i p t .

R E F E R E N C E SA n o n . ( 1 9 7 8 ) . B i o a s s a y p r o c e d u r e s f o r t h e o c e a n d i s p o s a l p e r m i t p r o g r a m . E P A - 6 0 0 / 9 - 7 8 - 0 1 0 .U S E n v i r o n m e n t a l P r o t e c t i o n A g e n c y , G u l f B r e e z e, 1 2 1 p p .B a hn e r , L . H . , W i l son , A . J . , j r , S he p pa r d , J . M . , P a t r i c k , J . M . , j r , G oo dm a n , L . R . & W a l sh , J . E .( 1 9 77 ) . K e p o n e b i o c o n c e n t r a t i o n , a c c u m u l a t i o n , l o ss a n d t r a n s f e r t h r o u g h e s t u a r i n e f o o d c h a i n s .C h e s a p e a k e S c i . , 1 8 , 2 9 9 - 3 0 8 .C lu t t e r , R . I . & T he i l a c ke r , G . H . ( 1971 ) . E c o log ic a l e f f i c i e nc y o f a pe l a g i c sh r im p : e s t im a te s o f

g r o w t h , e n e r g y b u d g e t , a n d m o r t a l i t y s t u d ie s . F i s h . B u l l . , 6 9 , 9 3 - 1 1 5 .John s , D . M . , B e r r y , W . J . & W a l ton , W . ( 1981 ) . I n t e r na t ion a l S tud y o n A r t e m i a . X V I . S u r v iva l ,g r o w t h a n d r e p r o d u c t i o n p o t e n t i a l o f t h e m y s i d , M y s i d o p s i s b a h i a ( M o l e n o c k ) f e d v a r i o u sg e o g r a p h i c a l c o l l e c t i o n s o f t h e b r i n e s h r i m p A r t e m i a . J . e x p . m a r . B i o l . E c o l . ( S u b m i t t e d f o rp u b l i c a t i o n . )K a l l e , K . ( 1971 ) . S a l in i t y : ge ne r a l i n t r oduc t ion . I n : M a r i n e e c o l o g y - 1 , p a r t 2 , e d . O . K inne .L o n d o n , W i l e y - In t e rs c i e n ce , p p . 6 8 3 - 8 .K i n n e , O . ( 1 9 7 6 ) . C u l t i v a t i o n o f m a r i n e o r g a n is m s : w a t e r - q u a l it y m a n a g e m e n t a n d t e c h n o l o g y . I n :M a r i n e e c o l o g y - 3 , p a r t 1 , e d . O . K i n n e . C h i c h e s t e r , J o h n W i l ey & S o n s , p p . 1 9 - 3 0 0 .K i n n e , O . ( 1 9 7 7 ) . C u l t i v a t i o n o f a n i m a l s -r e s e ar c h c u l ti v a t i o n . I n : M a r i n e e c o l o g y - 3 , p a r t 2 , ed .O . K i n n e . C h i c h e s t e r , J o h n W i le y & S o n s , p p . 5 7 9 - 1 2 9 3 .L ~ ge r , P h . & S o r ge loos , P . ( 1981a ) . I n t e r na t iona l s t udy on A r t e m i a . X X I I . S to r a ge o f A r t e m i ana up l i i f r om va r ious ge og r a ph ic a l o r ig in s i n t he r e f r i ge r a to r - po t e n t i a l s a nd lim i t s o f i t s a pp l i -c a b i l it y i n a q u a c u l t u r e h a t c h e r i e s . ( I n p r e p a r a t i o n . )L ~ ge r , P h . , V a nha e c ke , P . & S o r ge loos , P . ( 1981b ) . I n t e r na t iona l s t udy on A r t e m i a . X X I I I . N u t r i-t i o n a l v a l u e o f A r t e m i a na up l i i f r om v a r ious ge og r a ph ic a l o r ig in s f o r t he m ys id M y s i d o p s i s b a h i a( M o l e n o c k ) . ( I n p r e p a r a t i o n . )N im m o , D . R . , B a hne r , L . H . , R igby , R . A . , S he p pa r d , J . M . & W i l son , A . J . j r . ( 1977 ) . M y s i d o p s i sb a h ia , a n e s tua r ine spe c i e s su i t a b l e f o r l i f e - c y c l e t o x i c i t y t e s t s to d e t e r m i n e t h e e f f e c ts o f ap o l l u t a n t . I n : A q u a t i c t o x i c o l o g y a n d h a z a r d e v a l ua t io n , e ds F . L . M a ye r & J . L . H a m e l ink ,W a s h i n g t o n , A m e r i c a n S o c i e t y f o r T e s t i n g a n d M a t er i al s , p p . 1 0 9 - 1 6 .O lne y , C . E ., S c ha u e r , P . S ., M c L e a n , S . , Y o u L u & S im p son , K . L . ( 1980 ) . I n t e r na t ion a l s t udy onA r t e m i a . V I I I . C o m p a r i s o n o f t h e c h l o r i n a t e d h y d r o c a r b o n s a n d h e a v y m e t a l s i n f iv e d i f f e r e n ts t r a i n s o f n e w l y h a t c h e d A r t e m i a a n d a l a b o r a t o r y - r e a r e d m a r i n e f i s h . I n : T h e b r i n e s h r i m pA r t e m i a - 3 , e ds G . P e r soone , P . S o r ge loos , O . R oe l s & E . J a spe r s . W e t t e r e n , U n ive r sa P re s s ,p p . 3 4 3 - 5 2 .

S o r g e lo o s , P . ( 1 9 8 0 ) . T h e u s e o f t h e b r i n e s h r i m p A r t e m i a i n a q u a c u l t u r e . I n : T h e b r i n e s h r i m pA r t e m i a 3 , e ds G . P e r soo ne , P . S o r ge loos , O . R oe l s & E . J a spe r s . W e t t e r e n , U n ive r sa P r e s s,p p . 2 5 - 4 6 .S o r ge loos , P . & P e r soone , G . ( 1973 ) . A c u l tu r e sy s t e m f o r A r t e m i a , D a p h n i a a n d o t h e r i n v e r t e b r a t e s ,w i t h c o n t i n u o u s s e p a r a t i o n o f t h e l a rv a e . A r c h . H y d r o b i o l ., 7 2 , 1 3 3 - 8 .S po t t e , S . ( 1979 ) . S e a w a t e r a q u a r i u m s . N e w Y o r k , J o h n W i l e y & S o n s .

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