Lavens 1984 Aquacultural-Engineering

7/28/2019 Lavens 1984 Aquacultural-Engineering

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Aqua cu l rura l Eng ineer ing 3 ~1984) 22 I- 235

C o n t r o l l e d P r o d u c t i o n o f Artemia C y s t s U n d e r S t a n d a r dC o n d i t io n s in a R e c i r cu l a t in g C u l tu r e S y s t e m *

Patrick Lavens and tPatrick So rge loosA r t e m i a Re ference Center, State U niversity of G hent, J . Plateaustraat 22,B-900 Ghent. Belgium

A B S T R A C TA f l o w - t h ro u g h c u l tu re s y s t e m is d e sc ri b e d f o r t h e c o n t ro l le d p ro d u c t i o no f Artemia cys ts in 10 l i tre aquaria u sing 500/00 art i f icial seaw ater enric hedwi th Fe-E DT A as cu l ture m ed iu m , micron i sed ricebran as fo o d source andc y c li c f lu sh e s o f t h e cu l tu re s w i t h N rg a s a s i n d u c t i o n m e c h a n i sm f o rovipari ty . Th e cul ture wa ter is rec irculated over a ro ta t ing b io logicalcon tac tor , a se t tl i ng ta nk a nd a U V d i s in f ec t ion un i t . C ys t s are con thu t -ous ly harves t ed by f i l t ra t ion o f t he cu lture e ff luen t . Pr oduc t ion y i e ldsaverage 0 .5 g dry cys t s per day and per aquar ium.

I N T R O D U C T I O NT h e b r i n e s h r i m p A r t e m i a is u s e d a s a f a v o u r i t e s t u d y - o b j e c t f o r f u n d a -m e n t a l a s w e l l as a p p l i e d r e s e a r c h in n u m e r o u s d i s c ip l i n es o f t h eb i o l o g i ca l s c ie n c e s (C l eg g an d C o n t e , 1 9 8 0 ; S o r g e l o o s , 1 9 8 0 ) . T h em a i n r e a s o n f o r t h e p o p u l a r i t y o f A r t e m i a r e s u l t s f r o m i t s w o r l d w i d ea v a i l a b i li t y in t h e f o r m o f d r y c y s t s . P r e c is e i n f o r m a t i o n , h o w e v e r , o nt h e p r o d u c t i o n c o n d i t io n s o f th e s e d o r m a n t e m b r y o s , w h i c h in f a c t a rea l w a y s h a r v e s te d f r o m w i ld p o p u l a t i o n s , a r e m o s t l y u n k n o w n ; e .g .b i o t i c a n d a b i o t i c c o n d i t i o n s u n d e r w h i c h t h e c y s t s h a v e b e e n p r o d u c e d ,t i m e a n d p l a c e o f t h e ir d e p o s i t i o n , e tc . L a c k o f t h e s e e s s e n ti a l d a t a c a nb e a n i m p o r t a n t o b s t a c l e in t h e i n t e r p r e t a t i o n o f e x p e r i m e n t a l r e s u lt s

* Technique pro tected by Dutch Patent N o. 83201 335-3.t Senior Research Associate at the Belgian National Science Fou nd atio n (NFW O).

221Aquacu l tura l Eng ineer ing 0144-8609/84/$03.00 Elsevier Appl ied SciencePublishers Ltd, England, 1984. Printed in Great Britain


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222 Patr ick Lavens , Pa t r ick Sorge loos( G u n t h e r S i U e r o e t a L , 1 9 8 0 ; S o r g e l o o s . 1 9 8 0 ; V a l l e j o e t a L , 1 9 8 0 ;

V a n h a e c k e a n d S o r g e l o o s, 1 9 8 2) .I n t h i s r e g a r d t h e r e i s a c r i t i c a l n e e d f o r a c u l t u r e t e c h n i q u e t h a t

s h o u l d a llo w t h e p r o d u c t i o n o f c y st s u n d e r c o n t r o l l e d c o n d i t i o n s o fs a l i n i t y , t e m p e r a t u r e , l i g h t , f o o d q u a l i t y a n d q u a n t i t y , p o p u l a t i o nd e n s i t y , e tc . T h i s c o n t r o l l e d c y st p r o d u c t i o n t e c h n i q u e s h o u l d i n d e edp r o v i d e u n i q u e o p p o r t u n i t i e s t o s t u d y t h e s in gle a n d / o r c o m b i n e de f fe c t (s ) o f a n y a b i o t ic a n d / o r b i o t i c p a r a m e t e r o n t h e r e p r o d u c t i o nb i o l o g y a s w e l l as o n t h e c h a r a c t e r is t i c s o f t h e o f f s p r in g .

T h e t e c h n i c a l f e a si b il it y o f c o n t r o l l e d c y s t i n d u c t i o n h a s b e e nd e m o n s t r a t e d b y V e r s ic h e le a n d S o r g e l o o s ( 1 9 8 0 ) f o r l a b o r a to r yp o p u l a t i o n s c u l t u r e d u n d e r b a t c h c o n d i t i o n s i n a i r - w a t e r l i f t o p e r a t e dr a c e w a y s ( B o s s u y t a n d S o r g e l o o s , 1 9 8 0 ) . I n t h e m e a n t i m e , h o w e v e r ,m u c h p r o g r e s s h a s b e e n m a d e i n i n t e n s i v e A r t e m i a c u l t u r i n g u s i n gf l o w - t h r o u g h c u l t u r e t e c h n i q u e s a s d e s c r i b e d b y S h a r f s t e i n e t a l .( 1 9 7 9 ) , T o b i a s e t a l . ( 1 9 7 9 ) a n d B r i s s e t e t a l . (1 98 2 ) ; i.e . h igh surv iva lr a t e s c a n b e a s s u re d in c u l t u r e d e n s i ti e s o f m o r e t h a n 18 0 0 0 a n i m a l sp e r l i tr e . W e h a v e a d a p t e d t h e l a t te r f l o w - t h r o u g h t e c h n i q u e f o r re a ri n gh i g h d e n s i t i e s o f a d u l t b r in e s h r i m p in a c l os e d c u l t u r e s y s t e m a n d h a v ed e v e l o p e d a s i m p l e s t r e s s t e c h n i q u e w h i c h d o e s n o t r e d u c e p o p u l a t i o ns u rv iv a l b u t a ss ur es a m a x i m a l i n d u c t i o n o f t h e o v i p a r o u s m o d e o fr e p r o d u c t i o n .

D E S C R I P T IO N O F T H E C U L T U R E S Y S T EM

T h e c u l t u r e s y s t e m , 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 , e s s e n t i a l l y c o n s i s t so f t h e f o l lo w i n g c o m p o n e n t s : f o u r r e a ri n g u n i t s o f f iv e c u l t u r e a q u ar iae a c h ; a d i s t r i b u t i o n s y s t e m f o r t h e c u l t u r e w a t e r i n c l u d i n g a r e s e r v o i r ,h e a d t a n k , d i s i n f e c t i o n u n i t a n d s u p p l y p i p e ; a n e f f l u e n t d r a in a g es y s t e m w i t h o n e c y s t r e c u p e r a t i o n f i lt e r f o r e a c h c u l t u r e - u n i t ; a b io -l o g i c a l f i l t r a t i o n u n i t a n d a s e t t l i n g t a n k .

T h e a q u a r i a a re m a d e o f t r a n s p a r e n t P V C ( se e s p e c if ic d e s ig n inF i g. 2 ) a n d h o l d a c u l t u r e v o l u m e o f 1 0 l it re s e a c h. T h e y a r e g r o u p e d inf iv es in f o u r s e p a r a t e u n i t s , s o a s t o f o l l o w t h e c o m p a r a t i v e s t u d y o fd i f f e r e n t p a r a m e t e r s , e .g . t e m p e r a t u r e ( s e p a r a te w a t e r b a t h f o r e a c hu n i t) , f o o d q u a l i t y a n d / o r q u a n t i t y , l ig h t c o n d i t i o n s , e tc .

C u l t u r e w a t e r is p u m p e d u p f r o m a 3 m 3 r e s e r v o i r i n t o a 1 0 0-1 itreh e a d t a n k w h i c h is c o n n e c t e d w i t h a f l o w - t h r o u g h d i s i n f e c ti o n u n i t


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Controlled A r t e m i a cyst production in fTow-through culture system 2 2 3

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[ i f - l !Fig . 1 . Sch em atic d iagram of the rec ircu la t ing cul tu re sys tem for Arternia cys tp roduc t ion . (1 ) r e se rvo i r ; (2 ) pump ac t iva ted by leve l de tec to r (4 ) ; (3 ) head tank ;(5 ) UV s te ri li se r: (6 ) w a te r supp ly p ipe : (7 )a ir /N ,_ supp ly ; (8 )cu l tu re un i t con ta in ingf ive aqua r ia ; (9 ) d ra in gu t te r ; (10 ) co l lec to r t ank con ta in ing cys t r ecupera t ionf il te rs; (11 ) ro ta t ing b io logica l co nta c to r ; ( I 2) se t t l ing tank .

t h a t e n d s in t h e s u p p l y p i p e m o u n t e d a b o v e t h e a q u a ri a . T h e d i s i n fe c -t i o n s y s t e m ( s c h e m a t i c a l l y o u t l i n e d in F ig . 3 ) c o n s i s t s o f t w o U V t u b e s( P h iU i p s T U V 3 0 W ) a r o u n d w h i c h g la ss j a c k e t s w i t h in - a n d o u t l e tt u b e s h a v e b e e n f i x e d w i t h s i l i c o n e g l u e . O p t i m a l d i s i n f e c t i o n i s a s s u r e db y t h e f l o w o f c u l t u r e w a t e r t h r o u g h t h e 2 .5 c m i n t e r s p a c e w i t h i n t h et w o g la ss c y l i n d e r s t h a t e a c h h o l d a U V t u b e . W a t e r i n t a k e i n e a c ha q u a r i u m c a n b e a d j u s t e d w i t h t w o t a p s f i t t e d to t h e p la s t i c t u b i n gt h a t is c o n n e c t e d w i t h t h e s u p p l y p i p e . A s t h e f il li ng o f t h e h e a d t a n kis c o n t r o l l e d b y a t w o - le v e l e l e c t r o n i c d e t e c t o r , d i f f e re n c e s i n w a t e rp r e s s u r e i n t h e s u p p l y p i p e c a n b e m i n i m i s e d .

D r a i n a g e o f t h e c u l t u r e w a t e r ' s e f f l u e n t , l o a d e d w i t h fa e c a l p e l l e ts ,m o l t s , f o o d w a s t e s a n d e v e n t u a l l y c y s t s is a s s u r e d b y t h e c y l i n d r i c a ls t a n d p i p e f i l te r ( m e s h s iz es 3 0 0 , 4 0 0 , 5 5 0 o r 8 0 0 # m d e p e n d i n g o n t h el i f e s t a g e o f t h e Artemia i n c u l t u r e ) . T h e f i l te r s c r e e n is k e p t f r o mc l o gg i n g b y a c u r t a i n o f r i s in g a i r b u b b l e s p r o d u c e d b y t h e a e r a t i o nc o l l a r i n s t a l l e d a t t h e b a s e o f t h e s t a n d p i p e f i lt e r . A s a r e s u l t o f t h e


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Fig. 2. Schematic d iagram o f the cu lture aquarium. (A ) Lateral view; (B) upp erview. ( i ) w ater intake: (2) standpipe fi l ter w ith PVC ring (3) and aera tion collar (4);(5) drain with T-piece; (6) lid.

s p e ci fi c d e s ig n o f t h e a q u a r i a , t h e a e r a t i o n a s su re s m a x i m a l o x y g e n a t i o na n d c u l t u r e m i x i n g w i t h m i n i m a l h a r m t o t h e a ni m a ls . A n i m a l s t h a te v e n t u a l l y h a v e b e e n s k i m m e d o f f b y t h e a i r b u b b l e s c a n g e t b ac k in t ot h e c u l t u r e b y c o v e r in g t h e u p p e r p a r t o f t h e s ta n d p i p e f il te r w i t ha P V C - ri ng t h a t e x t e n d s u n t i l j u s t u n d e r n e a t h t h e w a t e r s u rf a ce . T h ed r a i n o f t h e s t a n d p i p e f i lt e r h o l d s a T - p i e c e t o a v o id o b s t r u c t i o n o ft h e e f f l u e n t f lo w b y a i rb u b b l e s . T h e a e r a t i o n r a te i n e a c h a q u a r i u mc a n b e a d j u s t e d w i t h a n a i rt a p c o n n e c t e d t o a c e n t r a l a ir d i s t r i b u t i o np i p e i n s t a ll e d a b o v e t h e a q u a r i a . T h e c e n t r a l a i r p i p e is c o n n e c t e d w i t ha n a ir b l o w e r a n d a N 2 g a s c y l i n d e r v ia a t h r e e - w a y e l e c t r i c va lv e .

T h e e f f l u e n t s f r o m t h e f i v e a q u a r i a o f e a c h u n i t a r e c o l l e c t e d i n t ot h e s a m e g u t t e r t h a t d r a i n s in t o o n e o f t h e f o u r c y s t r e c u p e r a t i o n f il te rs


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Controlled A r t e r n i a cyst production in flow-through culture sysrern 2 2 5

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Schematic diagram of the U V ster il iser. ( I ) UV tube: f2)glass- jacket ; (3)water inflow: 14) water outtlow.

i n s t a ll e d in a c o l l e c t o r t a n k . T h e s p e c i f i c d e s ig n o f t h e s e c y s t r e c u p e r a -t i o n f il te r s p r e v e n t s t h e ir f i l t e rs c r e e n c lo g g in g o v e r o p e r a t i o n a l p e r i o d so f a m i n i m u m o f 2 4 h.

T h e e f f l u e n t s f r o m t h e c o l l e c t o r t a n k a re d r a in e d i n to th e b i o l o g ic a lf i l t r a t io n u n i t. T h e p r i n c ip l e o f t h e r o t a t i n g b i o l o g i c a l c o n t a c t o r ( R B C )h a s b e e n s e l e c t e d b e c a u s e o f i ts s u c c e s s f u l u s e in fi sh a n d p e n a e i ds h r i m p c u l t u r i n g i n c l o s e d s y s t e m s ( L e w i s a n d B y n a k , 1 9 7 6 ; M o c ke t a l . , 1 9 7 7 ) a n d b e c a u s e o f i ts s ta b l e o p e r a t i o n u n d e r c o n d i t i o n s o ff l u c t u a t i n g h y d r a u l i c a n d o r g a n i c l o a d in g s ( C l ar k e t a l . , 1 9 7 7 ; A u t o t r o lC o r p . , 1 9 7 8 ) . T h e R B C s y s t e m ( s c h e m a t i c a l l y o u t l i n e d in F ig . 4 )c o n s i s t s o r 4 u n i t s o f 1 5 s a n d - b l a s t e d P V C - d i sc s ( 5 0 c m i n d i a m e t e r , 0 . 1c m t h ic k ) t h a t r o t a t e a t 6 r p m in f o u r i n t e r c o n n e c t e d c o m p a r t m e n t s o fa V - s h a p e d c o n t a i n e r . A t a n y t i m e 4 0 % o f t h e 2 5 m 2 d i s c -s u r f ac e iss u b m e r g e d in t h e e f f l u e n t . F l u o r e s c e n t l ig h t t u b e s a re i n st a ll e d j u s t


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226 Patr ick Lavens , Pa t r ick Sorge loos

[ g : . : : : : : . i i : : i i i = : : i i i i : . : : i :: i ! : : i : :i i i i il i i ii i i : : i :i i i i i i i :i i i i i i i } ! i L

E 5 5J l 4 6 I x 2 6 ,

Fig . 4 .C B

Schematic dia gra m o f the rotating biological conta ctor . (A) T op view;(B) lateral view; (C) view o f transverse s ection through one unit.

a b o v e t h e l as t R B C u n i t s o a s t o i n d u c e a lg al g r o w t h r e s u l ti n g i n N O 3r e m o v a l ( K e n n e d y , 1 9 8 0 ) . A s a r e su l t o f t h e s p e c if ic f l o w p a t t e r n o ft h e e f f l u e n t w a t e r t h r o u g h t h e f o u r R B C - c o m p a r t m e n t s se q u e n ti a ls u b s t r a t e c o n t a c t is o p t i m i s e d a n d m a x i m a l p u r i f ic a t i o n a s s u r ed . P r i o rt o i ts d r a i n a g e i n t o t h e c u l t u r e w a t e r r e s e r v o ir , t h e R B C e f f l u e n t f l o w si n t o a 5 0 0 l it re d e c a n t a t i o n t a n k f o r r e m o v a l o f t h e b i o f l o c c u l e s t h a te v e n t u a l l y s e p a r a t e f r o m t h e R B C d i s cs a n d a r e c ar r ie d a l o n g w i t h t h ep u r if ie d w a t e r . C o n c e n t r a t e d f o o d s u s p e n s i o n , e .g . m i c r o n i s e d r i c e b r a ns u s p e n s i o n , is d i s t r i b u t e d t o t h e i n d i v i d u a l c u l t u r e a q u a r i a v ia a m u l ti -c h a n n e l p e r i s t a lt i c p u m p w h i c h is a c t i v a t e d b y an a d j u s t a b l e e l e c t r o n i ct i m e r .


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Controlled Artemia cyst production in flow-through culture system 227PRACTICAL OPERATION OF THE CULTURE SYSTEM

Artificial seawater made up with six salts and tapwater, as advised bySorgeloos e t a l . (1983) for culturing A r t e r n i a from nauplius to adult,appeared to be a suitable culture medium for brine shrimp reproductionprovided the salinity was increased to 50%o by addition of saturatedNaC1 brine; i.e. A r t e m i a populations have been maintained in thepresent system over periods of several months (Lavens, 1981). Thefollowing abiotic parameters were analysed at weekly intervals in orderto evaluate the water quality changes: pH, BODs, NH3-N, NO2-N andNO3-N. Purification rates of the recirculation system approximated87%. NH3-N and NO2-N concent rations remained always below 0.01and 0-05 ppm respectively. The level of NO3-N, however, was graduallyincreasing but could be kept at a maximum of 2 ppm by a 20% waterchange every month. The pH fluctuated within the range 7-8-8-4.

Culture tests performed at different hydraulic retention times ofculture medium in the aquaria revealed that an optimal compromisebetween faeces/exuviae/cysts removal from the aquaria, feedingquantities and rate of clogging of the standpipe filters was achieved atflo* rates of 3 litres h -x per aquar ium, i.e. at a retention time of about3 h (Lavens, 1981).

Growth and survival tests with animal densities ranging from 10 to20 per mt revealed no significant differences; i.e. mortalit ies were nevermore than 20% and adult sizes were attained after the same cultureperiod of about three weeks; reproduction rates, however, were slightlyhigher at 10 adults per ml as compared to 20 per ml (Lavens, 1981). Inthis regard and also to minimise the risks of filter clogging with exuviaeand faeces, animal densities were kept at 10 per ml, i.e. 100000A r t e m i a per aquarium.

By analogy with the results of Versichele and Sorgeloos (1980),cyclic oxygen stresses in combination with Fe-EDTA addition to theculture medium have been used as an efficient induction mechanism foroviparous reproduction. The Fe-EDTA concentration in the culturemedium was kept at 30 ppm by regular addition of new solution to thereservoir. As compared to the aeration interruption technique ofVersichele and Sorgeloos (1980) much more efficient oxygen stressesin culture media of much lower satinities can be achieved with muchmore restricted impact on animal survival by the cyclic switch fromairbubbling to N2 gas flushing in the culture aquaria that are covered


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228 2arrick Lavens, Patrick Sorgeloosb y a li d. i.e . d e t a i l e d s t u d i e s r e v e a l e d t h a t m a x i m a l o v i p a r i t y i n d u c t i o nis a t t a i n e d b y a 4 m i n s w i t c h f r o m a i r t o N 2 a t 3 h in t e r v a l s ( L a v e n s ,1 9 8 1 ). A s c a n b e s e e n f r o m F i g. 5 t h e c y c l i c N 2 - t lu s h e s r e s u l t in v e r ye f f i c i e n t 0 2 s t re s se s w h i c h a r e o f b u t s h o r t d u r a t i o n . T h e Ar tem iap o p u l a t i o n s d o n o t a d a p t t o t h is c y c l i c t r e a t m e n t , r e m a i n in g o v i p a r o u so v e r p e r io d s o f s ev e ra l m o n t h s .

E v e r y d a y t h e c o n t e n t o f e a c h c y s t r e c u p e r a t i o n f i l te r is d r a i n e d o v e ra 1 50 ~ m s c r e e n ; t h e l a t t e r is r i n s e d t h o r o u g h l y w i t h t a p w a t e r s o a s t ow a s h o u t t h e s m a l l p a r t i c le s . C y s t s a re s e p a r a t e d f r o m t h e r e m a i n i n gs l u d g e ( e .g . f o o d p a r t i c l e s , f a e c a l p e l l e t s, e x u v i a e , e t c . ) b y a t w o s t e pd i f f e re n t i a l s e d i m e n t a t i o n p r o c e d u r e in b ri n e a n d t a p w a t e r a s d e s c r i b e db y S o r g e l o o s et aL ( 1 9 7 8 ) . C y s t s a r e f in a l ly s t o r e d in d e h y d r a t e d f o r mf o l lo w i n g o n e o r a n o t h e r t e c h n i q u e f o r d r y i n g o r d i a p a u s e - d i s a c t i v a t i o n( L a v e n s et aL , 1 9 8 4 b ) .

T h e f o l lo w i n g p r o c e d u r e s h a v e t o be p e r f o r m e d r o u t i n e l y in o r d e r t oa ss ure o p t i m a l m a i n t e n a n c e o f t h e c u l t u r e s y s t e m :- d a il y m o n i t o r i n g a n d e v e n t u a l a d j u s t m e n t o f t e m p e r a t u r e , a e r a t i o n

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Contro l l ed Artemia c y s t p r o d u c t i o n in f l o w - t h r o u g h c u l tu r e s y s t e m 229aquarium; standpipe filters are daily removed for cleaning (brushingand washing of filters);

- once a week all supply and outlet tubes, food distribution tubesas well as the tank holding the cyst collector filters are rinsedthoroughly;

- every month settled sludge and biofloccules are syphoned out ofthe RBC and the sedimentation tank; waterlosses are compensatedby adding tap water to the reservoir.

EXPERIMENTAL RESULTSIn one experiment A r t e m i a populations from Lavalduc (France,parthenogenetic) and Great Salt Lake (Utah, USA, bisexual), have beenkept in the present culture system for a period of two months (25C,50%0). Cultures were started with animals at different stages of theirlarval development, i.e. juveniles, preadults and reproductive adults.The reproductive characteristics of the Lavalduc and Great Salt LakeA r t e m i a in the different culture combinations are summarised in Table1. It appears from these data that in all cultures more than 80% of thefemales reproduce by oviparity. The results also reveal tile importanceof the larval stage at which induction is started. Indeed, only thoseanimals that were already induced at their juvenile stage gave oviparousoffspring as first brood whereas those transferred in the preadult oradult stage into the cyst production culture system only produced cystsfrom their second brood onwards. The present observation appears tocontradict with the theory of D'Agostino (1980) that tt~e first broodof A r t e m i a is invariably ovoviviparous. Our findings furthermoreindicate that a certain period of time is required for the shift tooviparity, which might reveal the need for a specific substance, whicheventually should be present in but a small concentra tion to induce cystproduction in brine shrimp. Since haematin, in fact a decompositionproduct of haemoglobin, is an essential component of the cyst shell(Gilchrist, 1960) the key product could be a specific haemoglobin. Theearlier reported pathway of haemoglobin-induced oviparity (Dutrieu,1960; Versichele and Sorgeloos, 1980) can indeed be further unravelled,i.e. low oxygen levels stimulate the p roduct ion of hemoglobins ingeneral (Bowen e t a l . , 1969) but especially enhance the synthesis of onespecific haemoglobin Hb-III (Heip e t a l . , 1978; Van den Branden e t a l . ,


10/15

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11/15

Controlled Artemia cyst prod uctio n in ITow-through culture system 2311 9 7 8 ; H e i p e r al . , 1 9 8 0 ) , t h e s y n t h e s i s o f w h i c h , a c c o r d i n g t o G r ig l i at ti( 1 9 6 9 ) , is d e p e n d e n t o n th e c o n c e n t r a t i o n o f ir o n in t h e m e d i u m .P r e l i m i n a r y o b s e r v a t i o n s o f s i g n i f i c a n t i n c r e a s e s i n t h e H b - I I I c o n -c e n t r a t i o n u p t o 13% o f th e t o t a l h a e m o g l o b i n c o n t e n t in p r e -a d u l tA r t e m i a u p o n t h e i r tr a n s f e r in t h e p r e s e n t c u l t u r e s y s t e m a p p a r e n t l yc o n f i r m t h e h y p o t h e s i s o f H b - I II r e g u l a t i o n o f o v i p a r i t y in a d u l tA r t e m i a ( L a ve ns , 1982) .

A n o t h e r p r o o f t h a t h a e m o g l o b i n s 'i n g en e ra l ' d o n o t c o n t r o l th em o d e o f r e p r o d u c t i o n in A r t e m i a r e s u lt s f r o m o u r o b s e r v a t i o n s o f r e dA r t e m i a t h a t c o n t i n u o u s l y p r o d u c e n a u p l i i a n d o f p i n k - re d o v i p a r o u sa n im a l s. A c c o r d i n g t o C h o w ( 1 9 6 8 ) a n i m a l c o l o r a t i o n is i n d e e d o n l ya c r u d e w a y t o e s t i m a t e h a e m o g l o b i n c o n c e n t r a t i o n s . In t h i s r e g a rd t h ep r o p o s e d c o r re l a ti o n b e t w e e n a n i m a l c o lo r a t io n a n d m o d e o f r e p r o d u c -t i o n , a s p r o p o s e d b y V e r s i c h e le a n d S o r g e l o o s ( 1 9 8 0 ) , s h o u l d b er e c o n s i d e r e d .

P R O D U C T I O N Y I E L D ST h e p r e s e n t c u l t u r e s y s t e m ha s n o t b e e n o p t i m i s e d y e t s o a s t o m a x i m i s ep r o d u c t i o n y i e l d s . N o n e t h e l e s s w i t h a p o p u l a t i o n o f p a r t h e n o g e n e t i cL a v a l d u c A r t e m i a c u l t u r e d o n m i c r o n i s e d r i c e b r a n a t 2 5 C s u s t a i n a b l ey i e l d s o f 0 . 5 g d r i e d c y s t s p e r a q u a r i u m ( 1 0 0 0 0 0 a n i m a l s) a n d p e r d a yh a v e b e e n a c h i e v e d . I n c r e a s e d p r o d u c t i o n r e su l ts c a n b e e x p e c t e d f ro md i e t m a n i p u l a t i o n s ; e.g . b a se d o n t h e o b s e r v a t i o n s o f P r o v a s o l i a n dP i n t n e r ( 1 9 8 0 ) t h a t l i p i d s a r e e s s e n t i a l f o r t h e c o n t i n u e d f e r t i l i t y o fA r t e m i a , w e e n r i c h e d t h e r i c e b r a n w i t h 9 % o il a n d n o t i c e d n o t o n l yi n c re a s e d f e r ti l is a t io n r a te s b u t e s p e c i al ly a m u c h h i g h e r f e c u n d i t y r a tep e r f e m a l e ( L a v e n s e t a l . , 1 9 8 4 a ) .

C O N C L U S I O N ST h e p r e s e n t c u l t u r e s y s t e m h a s p r o v e n t o b e a r e li a b le t e c h n i q u e f o r t h ec o n t r o l l e d p r o d u c t i o n o f A r t e m i a c y s t s , i.e . w i t h b r i n e s h r i m p ( f r o mL a v a l d u c , F r a n c e ; G r e a t S a l t L a k e , U t a h , U S A ; S a n P a b l o B a y , C a l i f o r n i a ,U S A ; T i e n t s i n , C h i n a ; a n d C h a p l i n L a k e , C a n a d a ), a d o m i n a n t ( > 9 0 % )o v i p a r o u s r e p r o d u c t i o n w a s m a i n t a i n e d c o n t i n u o u s l y . A s e a r l i e r s t a t e db y V e r s i c h e le a n d S o r g e l o o s ( 1 9 8 0 ) ' . . . t h e k e y t o s uc c es s is t h e


12/15

232 Patr ick Lavens , Pa t r ick Sorge loosa p p l i c a t i o n o f c y c l i c o x y g e n s t re ss es in m e d i a e n r i c h e d w i t h F e - E D T A ' .A s c o m p a r e d t o t h e b a t c h t e c h n i q u e o f t h e l a tt e r a u t h o r s th e n e wt e c h n i q u e i n f l o w - t h r o u g h a q u a r i a w i t h c y c l i c N : f l u s h e s o f f e r s s e v e r a li m p o r t a n t a d v a n t a g e s :- m u c h h i gh e r d e n si ti es o f A r t e m i a c a n b e m a i n t a i n e d f o r l o n g e r

p e r i o d s o f t i m e a t r e d u c e d m o r t a l i t y r a te s , r e s u l t i n g ha i n c r e a se dy i e l d s p e r c u l t u r e v o l u m e ;

- s i m p l e , e f f i c ie n t a n d c o n t i n u o u s h a r v e s ti n g o f ti le p r o d u c e d c y s t s;- m u c h m o r e e f f i c i e n t o x y g e n s tr es se s a t r e d u c e d s a li n it ie s w i t h

m i n i m a l in t e r f e re n c e o n o t h e r c u l t u r e c o n d i t i o n s .T h i s n e w t e c h n i q u e f o r c o n t r o l l e d c y s t p r o d u c t i o n o f f e rs ver?. i n te r e st -in g o p p o r t u n i t i e s f o r d i ff e r e n t c o m p a r a t iv e s tu d i es , fo r e x a m p l e :- d e t e rmina t i on o f s t r a i n - s p e c i f i c ( g e n o t y p i c a l ) c h a r a c t e r i s t i c s , i . e .

s t u d y o f t h e i n f l u e n c e o f (a ) b i o t i c f a c to r s o n t h e c h a r a c t e ri s ti c s o fc y s ts a n d t h e i r p r o g e n y - b a se d o n t h e re s u lt s o b t a i n e d , s p e ci fi cc r o s s- b r e ed i n g t e s ts c o u l d b e c o n s i d e r e d fo r t h e p r o d u c t i o n o f n ewv a r i e t i e s o f A r t e m i a w i t h a t t r a c t i v e f e a t u r e s f o r s p e c if ic u s e s ina q u a c u l t u r e ;

- t h e e ff e c t o f p r o d u c t i o n c o n d i t i o n s a n d / o r p ro c e ss in g m e t h o d s o nt h e i n h i b i t i o n o f t h e d i a p a u s e m e c h a n i s m in t h e c r y p t o b i o t i c c y st s( L a v e n s e t a l . , 1 9 8 4 b ) ;

- t h e i n f l u e n ce o f p r o d u c t i o n c o n d i t i o n s o n t h e n u t r i t i o n a l q u a l i t y o ft h e c y s t s (n a u p l i i ) f o r s p e c i f i c p r e d a t o r l ar v ae ( L a v e n s e t a l . . 1 9 8 4 a ) ;t h r o u g h u p s c a l i n g o f t h e p r e s e n t t e c h n o l o g y , t h is c o u l d l ea d to t h ec o n t r o l l e d p r o d u c t i o n o f s ta n d a r d r e fe r en c e A r t e m i a c y s ts ( S i m p s o ne t a l . , 1 9 8 0 ; S o r g e l o o s , 19 8 1 ) w h i c h w ill b e o f c o n s t a n t a n d e v e n t u a l l ys u p r a n a t u r a l q u a l i t y ( e . g . f a t t y a c i d p r o f i l e ) .

A C K N O W L E D G E M E N T ST h i s s t u d y h a s b e e n r e p o r t e d b y a sc h o l ar sh i p f ro m t h e I n s t i tu t e f o rS c i e n ti f i c R e s e a r c h f o r t h e P r o m o t i o n o f I n d u s t r y a n d A g r i c u l tu r e( IW O N L ) t o P a t r i c k L a v e n s a n d b y a r e s e a rc h g r a n t f r o m t h e B e lg i anN a t i o n a l S c i en c e F o u n d a t i o n ( N F W O ) t o P a tr ic k S o rg e l o os .


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Contro l l ed Ar te mia c y s t p r o d u c t i o n in f l o w - t h r o u g h c u l tu r e s y s t e m 233R E F E R E N C E S

Au to t ro l Co rp . ( 1 9 7 8 ) . Au to t ro l W a s te wa te r t r e a tme n t sy s t e ms . Des ign manua l .Au to t ro t Co rp o ra t io n .Bossuy t . E . & Sorge loos. P . (1980) . Techno log ica l aspec ts o f the ba tch cu l tu r ing o fArtemia in h igh densi t ies . In : T h e B r i n e S h r i m p Artemia . Vol . 3 : Eco logy ,Cu ltur ing. Use in A qua cul tu re . eds G . Persoone, P . Sorgeloos , O. Roels andE. Jaspers, Universa Press. Wetteren, Belgium, pp. 133-52.

B ow en. S. T. . Leb herz , H. G.. P oo n. M .-C., C ho w , V. H. S. & G rigliatt i , T. A. (1969 ).T h e h e mo g lo b in s o f Ar temia sa l ina I. De te rmin a t io n o f p h e n o ty p e b y g e n o ty p ea n d e n v i ro n me n t . C o m p . B i o c h e m . P h y s i o l . , 3 1 , 7 3 3 - 4 7 .

Brisset, P. , Versichele, D., Bossuyt. E. , De Ruyck, L. & Sorgeloos, P. (1982). Highdens i ty f low- th rough cu l tu r ing o f b r ine sh r imp A r t e m i a o n i n e r t f e e d s - p r e -l iminary resu l t s wi th a modi f ied cu l tu re sys tem. Aquacu l tura l Eng ineer ing , 1,115-19 .

Chow. V. (1968) . Phys io logy o f A r t e m i a : e f f e c t o f n u t r i t i o n a n d o x y g e n t e n s io n o nsurvival and h em oglo bin pro du ct io n. Thesis , San F ransisco Sta te C ollege .

Clark, J . W., V iessm an, W. & Ha m m er, M. (Ed s) (1977 ). Biological-disc process. In :Water Supp ly ' and Po l lu t ion Con tro l , Harper and Row, New York pp . 557-62 .

Clegg. J . S . & C onte , F . P . (198 0) . A review of the ce llu lar and deve lopm ent b io logyo f A r t e m i a . In : T h e B r i n e S h r i m p A rtemia . Vol . 2 : Phy siology , Bioc hem istry ,Molecular Biology, eds G. Persoone, P. Sorgeloos, O. Roels and E. Jaspers,Universa Press, Wetteren pp. 11-54.

D'Agost ino. A. S . (1980) . The vi ta l requirements of A r t e m i a : phys io logy andnu t r i t ion . In : T h e B r i n e S h r i m p Artem ia . Vol. 2 : Phys io log y , Biochemis t ry ,Molecular Biology, eds G. Persoone, P. Sorgeloos, O. A. Rods and E. Jaspers,Universa Press, Wetteren.

Du tr ieu . J . (196 0) . O bservat ions b iochim iques e t physiolog iques sur le d& etoppe-m e n t d 'Ar temia sa l ina Leach . Arch . Zoo l . exp . gdn . , 99, 1-134.Gi lchr i s t , B . M. (1960) . Growth and fo rm of the b r ine sh r imp . Ar tem ia salina (L) .Proc. zool . Soc . Lond. . 134 (2) , 221-35.

Gr ig l ia tt i, T . A. (1969) . Di f fe ren t ia l indu c t ion o f hem oglob in syn thes is in A r t e m i asalina. Thesis, San Fransisco State College.

Gunther Sillero, M. A., Burillo, S. L. , Dominguez, E. , Olalla, A., Osuna, C. , Renart.J . , Sebastian, J . & Sillero, A. 11980). Multiple proteolytic enzymes in A r t e m i a .In : T h e B r i n e S h r i m p Artem ia , V ol . 2 : Phys io logy , B iochem is t ry , Molecu la rB iology , eds G. Perso one , P. Sorgelo os, O. Roels an d E. Jaspers, U niversa Press,Wet te ren , pp . 345-54 .

He lp , J. , Moens, L . & K ond o , M . (1978) . E f fec t o f con cen t ra t ions o f sal t andoxygen on the syn thes is o f ex t race l lu la r l l emoglob ins dur ing deve lopment o fA r t e m i a s a li n a D e v l B i o l . , 6 3 , 2 4 7 - 5 1 .


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234 Patrick Lavens, Patrick SorgeloosHeip, J . , Moens, L . , Hertsens , R. , Wood, E . J . , Heyl igen , H. , Van Broekhoven , A. ,

Vr in ts , R . , de C haf f oy , D. & Ko nd o , M. (1980) . Ar tem ia ex t race l lu la r hem o-g lob ins : on togeny , s t ruc tu re and in vivo radiolabel ing. In : The Brine ShrimpA rtemia , V ol . 2: Phy s io logy , Biochem is t ry , Molecu la r Bio logy , eds G. Persoone ,P . Sorgeloos , O. Roels and E . Jaspers , Universa Press , W etteren , pp . 4 27 -4 8.

K en ned y , R. M. (1980) . Recyc l ing and reuse o f w a te r fo r in tensive cu l tu re o fsh rimp . Re p o r t , Un iv e r s it y o f Ho u s to n .

Lavens , P . (1981) . S tud ie van de inv loed van d ive rse kon d i t ie s op de kenm erken vano v o v i v i - e n o v ip are n a k o m e l in g e n v a n Artemia g e p ro d u c e e rd in n a u wk e u r iggeko n t ro lee rde kw eeksys tem en . IWONL - Akt iv i te i tsve r lag .

Lavens , P . (1982) . S tud ie van de inv loed van d iverse kond i t ie s op de kenm erkenvan ovovivi - en ovipare n ako m eling en van Artemia geproduceerd in nauwkeur iggek ontro leerd e kw eek syste m en. IW ON L - Aktiv i te itsvers lag .

Lavens, P ., IAger, Ph. & Sorgeloos, P . (1984a) . M anipu la t ion of the fa t t y ac idprof i le in Artemia offsp r ing in a con t ro l led p ro duc t ion u n i t . (In p repara t ion . )

Lavens, P . , Tackaer t , W. & Sorgeloos, P . (1984b) . The inf luence of abiot ic andb io t i c c o n d i t i o n s o n th e h a t c h a b i l i t y a n d d i a p a u se -d e a c t iv a t io n o f Artemia-cys ts p rod uce d in a s tandard cu l tu re sys tem . ( ln p repara t ion . )

Lewis , W. M. & Bynak, G. L . (1976) . Evaluat ion of a revolving pla te type b iof l l terfo r use in rec i rcu la ted f i sh p ro duc t ion and ho ld ing un i t s . Trans. Am. Fish. Soc.,6 , 7 0 4 - 8 .M ock, C. R . , R oss, L . R. & Salser, B. R. (1977 ) . Design an d pre l imin ary evalu at ionof a c losed system for shr imp cul ture . In : Proc. 8th Ann. Meeting WMS, ed.J . W. A vault , J r , L ouis iana Sta te U nivers i ty , B aton Rou ge, pp . 335 -69.

Provasoli , L . & Pintner , I . J . (1980) . Biphasic par t icu la te m edia for the pa r theno -genet ic Artemia of S~te . In : The Brine Shrimp Artem ia , Vol. 2 : Phys io logy ,Biochemistry , Molecular Biology, eds G. Persoone, P . Sorgeloos , O. Roels andE. Jaspers, Universa Press, Wetteren, pp. 231-8.

Sharfs te in , B. A. , Tobias , W. J . & Roels , O. A. (1979) . The technical feasib i l i tyof mass cu l tu r ing o f the b r ine sh r imp Artemia sp. in the St-Croix artif icialupwel l ing sys tem. I I . Maximum s tock ing dens i ty and y ie lds and g rowthef f ic iency in f low- th rough raceway cu l tu re . In : Artificial Upwelling: Cultivationof the Brine Shrimp Ar te mia , Progress Report 1978-1979, Un ivers i ty of Texas.

Simpson, K. L . , Beck. A. D. & Sorgeloos, P . (1980) . Report workshop. I . Charac-t e r i z a t io n o f Arremia s t ra ins fo r app l ica t ion in aquacu l tu re . In : The BrineShrimp Artem ia , Vol . 3 : Eco logy , Cu l tu r ing , Use in A quac u l tu re , eds G. Persoone ,P . Sorgeloo s , O. Roels and E. Jaspers , U niversa Press, We tteren , pp . 409 -11 .Sorgeloos , P . (1980) . The use of the br ine shr imp Artemia in aquacu l tu re . In : TheBrine Shrimp A rtemia , V ol. 3 : Eco logy, Cultur ing, Use in A qua cul ture , edsG. Persoone, P. Sorgeloos, O. Roels and E. Jaspers, Universa Press, Wetteren,p p . 2 5 -4 6 .


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Controlled A rt em i a cyst production in flow-through culture system 235Sorgeloos, P . (19 81 ). Av ailabi l ity o f referen ce Artemia cysts . Aquaculture. 23 ,

381-2 .Sorgeloos, P. , Persoone, G. , Baeza-Mesa, M., Bossuyt , E. & Bruggeman, E. (1978).

The use of Artemia cysts in aquac ul ture: the con cept of 'ha tchin g ' and descrip-t ion of a new m eth od of cys t p rocessing . In : Proc. 9th Ann. Meeting IMS, ed.J . W. Av aul t , Jr . , L ouisiana State Un iversi ty , Baton Rou ge, pp. 71 5-21 .

So rgeloo s, P. , Bo ssu yt , E., Lavens, P. , l_~ger, Ph., Va nh aec ke , P. & Versichele, D.(1983 ). The use of the brine shrimp Artemia in crustacean hatcheries andnurseries. In: CRC Handbook ofMaric,dture. Vol . I : Crustacean A qua cul ture ,ed. J . P. McVey, CRC Press, Boca Ra ton. pp. 71-9 6.

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Lavens 1984 Aquacultural-Engineering

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