Leger 1983 Aquacultural-Engineering

7/28/2019 Leger 1983 Aquacultural-Engineering

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Aqua cu l t ura l Eng i neer ing 2 (1983) 6 9 -78

I n t e r n a t i o n a l S t u d y o n A r t em i a * X X I V . C o l d S to r a g e o fL i v e A r t e m i a N a u p l ii f ro m V a r i o u s G e o g r a p h i c a lS o u r c e s : P o t e n t i a l s a n d L i m i t s i n A q u a c u l t u r e

Philippe L6ger~-, Pau l Van haecke and Patrick Sorg eloos$A r t e m i a Re ferenc e Center, State U niversi ty o f Gh ent , J . Plateaustraat 22,

B-9000 Ghent , Belgium

A B S T R A C TFresh l y -ha t ched Artemia nau pli i fro m various geograp hical sources surviveds torage in a re fr igerator (2-4 C ) at d ens i t ies o f 2 00 0 per ml and above .E x c e p t f o r Artemia f ro m Chaplin L ake an d Buen os Aires , naupl iar v iabi li t ywas very high e ven a f ter 4 8 h storage, an d did n ot decrease signi f ican t lyaf ter a 24 h po s t - s torage t ransfer to 25C. Nei th er the naupl iar dry w eightnor b iochem ical co mp osi t ion change d s igni fi cant ly dur ing re fr igeration form ost s t rains tes ted . Com parat ive cul ture-tes t s wi th s tor ed and f reshly-h a t c h e d n a u p l i i a s J o o d f o r / u v e n i l e m a r in e m y s id s Mysidopsis bahia M.and larval carp Cyprinus carpio L. revealed s imi lar product ion per form-at lces .

I N T R O D U C T I O NA r t e r n i a is a p r a c t i c a l a n d s u i t a b l e l ar v a f o o d f o r b o t h m a r i n e a n d f re s h -w a t e r c r u s t a c e a n s a n d f i s h es ( K i n n e , 1 9 7 7 ) . B r in e s h r im p a r e c o m m o n l yu s e d a s f r e s h l y - h a t c h e d n a u p l i i, b e c a u s e o l d e r u n f e d n a u p l i i l o se t h e i rn u t r i t i o n a l v a l u e ( M o r r i s , 1 9 5 6 ; M a d d o x a n d M a n z i , 1 9 7 6 ; W a t a n a b e*Intern at ional interdisciplinary stud y on A r t e m i a strains coordinated by theA r t e m i a Reference C enter , State Univers ity of Gh ent , B elgium.t Research Assistant a nd :~Research Associate at the Belgian National ScienceFoundat ion (NFWO) . 69

A q u a c u l t u r a l E n g i n ee r in g 0 1 4 4 - 8 6 0 9 [ 8 3 / 0 0 0 2 - 0 0 6 9 [ $ 0 3 . 0 0 - Applied Science


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70 P. L~ger, P. Vanhaecke, P. Sorgelooset al., 1 9 7 8 ; D y e , 1 9 8 0 ) . I t is n o t c l e a r i f t h is is d u e t o b i o c h e m i c a lc h a n g e s ( B e n i j t s et al., 1 9 7 6 ) , c r it ic a l s iz e i n c r e a s e ( H e n t s c h e l , 1 9 6 8 ;S m i t h , 1 9 7 6 ) o r i n c r e a s e d s w i m m i n g a c t iv i t y ( M il l er et al., 1 9 7 9 ) .A l t h o u g h i t is o b v i o u s th a t f r e s h l y - h a t c h e d n a u p l i i a r e a p r e f e r r e d f o o d ,l it tl e a t t e n t i o n h a s b e e n p a i d t o t h e in s t a r -s t a g e a t w h i c h Artemia l a r v aea re o f f e r e d t o t h e p r e y i n g la rv a e . T h e o p t i m a l u s e o f Artemia n a u p l i i a sa s o u r c e o f li ve f o o d i m p l i e s t h e d a i l y i n c u b a t i o n o f c y s t s a n d h a r v e s t-in g o f n a u p li i . M e t h o d s o f o p t im i z i n g Artemia u s e a n d r e d u c i n g c y s t s -h a t c h i n g o p e r a t i o n s b y s t o r in g f r e s h l y - h a t c h e d n a u p l ii a t l o w t e m p e r a -t u r e w a s t h e r e f o r e s t u d i e d .

M A T E R I A L S A N D M E T H O D ST h e Artemia s t ra i n s u s e d i n th e e x p e r i m e n t s a r e l i s te d i n T a b l e 1 .C y s t s w e r e i n c u b a t e d u n d e r o p t i m a l h a t c h i n g c o n d i t i o n s ( S o r g e l o o s ,

T A B L E 1Percent Survival o f Artemia Naupli i from Different Geographical Sources Stored at2-4C for 24 h and 48 h at Densit ies o f 2 000 or 8 000 per ml

Source of Artemia cysts 24 h 48 h2-4C 2_4C

Macau (Brazil ) batch no. 87117 2 96.3 96.1Macau (Brazil) batc h no. 9710 51 94-0 91-4Macau (Brazil) ba tch no. 971 051 - 8 00 0 naupli i/ml 92-6 91-8San Francisco Ba y (San Francisco, USA ) ba tch no. 259 6 93"0 86-7*Shark Ba y (Australia) 94.5 93 "9San Pab lo Ba y (San Francisco, USA ) ba tch no. 1628 99.1 99-1Tientsin (People ' s Rep ubl ic of China) 100 97.0Reference Artemia cysts (Sorgeloos, 1981 ) 94.0 93-0Grea t Sal t Lak e (Uta h, USA ) 95 95-1Chaplin Lake (Canada) 12-4** 7-1 **Bue nos Aires (Argentina) 71.6* * 73-7 "*Lavalduc (Fra nce ) 97-8 95-6

*Significant ly differe nt at the level 0.05 .** Significantly diff ere nt a t the level 0-01.


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In ternat ional s tudy on Artemia - X X I V 711980), and a homogenous instar-I population was harvested afterdifferent cyst incubation periods (Vanhaecke and Sorgeloos, 1982a).The nauplii were separated from the hatching debris in a cylindricalseparator box (Persoone and Sorgeloos, 1972), rinsed with chilledseawater (2-4C), concentrated at densities of 2000 or 8000 nauplii perml seawater, and transferred to cylindro-conica l vessels (150 ml conten t).These were placed in a (thermostatic) refrigerator (2-4C) and con-stantly aerated. The percentage survival of nauplii was calculated bysubsampling from a uniform suspension with an automatic micropipetteafter 24 h and 48 h. Extra subsamples of a few strains were removedand held for 24 h at 25C in petri dishes after which time survival of thenauplii was determined. The instar-stages were determined according toHentschel (1968).

Dry-weight analyses were performed by the procedure of Vanhaeckeand Sorgeloos (1980), and determinations of total lipid were carriedout according to the method of Schauer and Simpson (1978). Separa-tion of fatty acid methyl esters was performed by gas chromatographyon a glass column (1 .80 m 2 mm I.D.) packed with 10% Altech CS-8on Chromosorb W-AW 100-120. Gas chromatographic conditions were:200C isothermal; carrier gas N: at 40 ml/min; F.I.D.-detection. Identi-fication and quantification was performed using a calibrated method ona HP3 390A plotter- integrator. Nutritional evaluation tests were:carried out according to the standard culture test procedure withM y s i d o p s i s b a h i a M. (L6ger and Sorgeloos, 1982a) and the culture testwith C y p r i n u s c a r p i o L. (Vanhaecke and Sorgeloos, 1982b).

Except for the fatty acid analysis, all data were treated statistically ina one way analysis of variance. Duncan's Multiple Range Test was usedto determine significant differences among means. Before analysis,survival data were normalized through an arcsin~/% transformation(Snedecor and Cochran, 1967).

RESULTS AND DISCUSSIONFor most A r t e m i a strains tested, storage of nauplii in densities up to8000 per ml at 2-4C did not significantly ( P > 0-05) affec t theirsurvival (Table 1). The high mor tal ity rates found for Chaplin Lake andBuenos Aires A r t e m i a confirm earlier findings that nauplii from the


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72 P . L ~ g e r , P . V a n h a e c k e , P . S o r g e l o o ss a m e s t r a in s d o n o t s u r vi v e w e l l a t 2 0 C a n d 3 0 C ; t h i s a p p e a r s t o b ec o r r e l a t e d w i t h l o w e r c a l o r ic c o n t e n t s o f t h e n a u p l i i o f t h e s e t w os t r a i n s ( V a n h a e c k e e t a l . , 1 9 8 2 ) .A f t e r 4 8 h s t o r a g e , t h e n a u p l i i w e r e s ti ll a t t h e i n s t a r I s ta g e . U p o nt r a n s f e r t o 2 5 C t h e i r m e t a b o l i s m w a s e n h a n c e d a n d m o l t in g in t o i n s ta rI I a n d I I I s ta g e s o c c u r r e d , a p p a r e n t l y w i t h o u t a n y s i g n i fi c a n t ( P > 0 . 0 5 )e f f e c t o n t h e i r s u r v iv a l ( T a b l e 2 ) .

A f t e r 2 4 h o f s t o ra g e a t 2 - 4 C , t h e n a u p l i a r d r y - w e i g h t l os se s w e r ei n s i g n i f i c a n t ( P > 0 - 0 5 ) . A f t e r 4 8 h s t o r a g e , t h e d i f f e r e n c e s w e r e st il lv e r y s m a l l a s c o m p a r e d w i t h t h e d r y w e i g h t l o ss e s a f t e r 2 4 h a t 2 5 C( T a b l e 3 ) .

A s c o m p a r e d t o a 26 % d e c r e a s e i n t o t a l l i pi d c o n t e n t w h e n k e p t a t2 5 C f o r 2 4 h ( B e n i j t s e t a l . , 1 9 7 6 ) , A r t e m i a n a u p l i i a p p a r e n t l y d i dn o t m e t a b o l i z e t h e i r l ip id r e s e r ve s w h e n s t o r e d a t 2 - 4 C , a n d t h e f a t t ya c id p a t t e r n s w e r e n o t s i g n if i c a n tl y c h a n g e d d u r i n g c o o l s t o ra g e( T a b l e 4 ) .

T h e s u rv iv a l, g r o w t h a n d r e p r o d u c t i v e d e v e l o p m e n t o f m y s i dj u v e n i le s w e r e a t l ea s t a s g o o d w h e n f e d n a u p l i i s t o r e d f o r 2 4 - 4 8 h a sw h e n f e d f r e s h l y - h a t c h e d A r t e m i a n a u p l i i ( T a b l e 5 ) .

T A B L E 2Percent Survival of Artemia Nauplii from D iffere nt Geographical Sources Stored at25C for 24 h Post-storageSource o f Artemia Storage conditions

2 4 h , 2 -4 C 2 4 h , 2 -4 ~ C+24 h, 25C

4 8 h , 2 -4 C 48 h , 2 -4 C+24 h, 25C

Macau, no . 971051 94-0 93.4 91-4M acau, no . 971051 92-6 94-5 91-8(8 000 nauplii per m l)Reference Artemia cys ts 90.6 89.2 88-4Shark Bay 94-5 94.8 93-9San Pablo Bay, no . 1628 99.1 97-0 99.1Tientsin 100 95.8 97

89 "588 "687 "894"210 094"1


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I n t e rna t i ona l s t udy on Artemia X X I V 73T A B L E 3Individual Dry W eights (/2g) of A r t e m i a Nauplii from D ifferent Geographical Sources

Stored Un der Various ConditionsS o u r c e o f Artemia Fresh ly - S t o red naup l i i

h a t c h e dn a up lii 2 4 h , 2 - 4 C 4 8 h , 2 - 4 C 2 4 h , 2 5 C a

Great Sa lt Lake 2 .42 2 - 36 -2 -5% ) 2 .22* ( -8 .0% ) 1 -59 ( -34-3% )San Pablo B ay, n o . 1 6 2 8 1 . 9 2 1 -8 7 (2 . 6 % ) 1 .7 5" ( -8 -0 %) 1 . 3 6 ( -2 9 . 2 % )Tients in 3 -0 9 3 - 0 2 -2 . 3 % ) 2 .8 5* ( -7 -8 %) 2 -3 7 ( -2 3 . 3 % )* Significantly diffe rent at the 0.05 level.a From Vanhaecke et al . (1982).

C a r p l ar v ae g re w w e l l w h e n f e d f r e s h l y - h a t c h e d a n d 2 4 h s t o r e dA r t e m i a n a u p l i i ( T a b l e 6 ) . T h e g r o w t h r a t e w a s , h o w e v e r , 8 % l o w e rw h e n f e d 4 8 h s t o r e d n a u p l i i ; a f t e r t h i s l o n g s t o r a g e , t h e n a u p l i i h a dp r o b a b l y l o st r e s i s t a n c e t o s u rv i v e t h e e x t r a s t re s s c r e a t e d b y t h et r a n s f e r i n t o f r e s h w a t e r a n d d i e d o f f r a p i d l y . T h e s u r v iv a l o f c a r p w a sn o t a f f e c t e d b y s t o ra g e c o n d i t i o n s o f t h e A r t e m i a ( T a b l e 6 ) .

C O N C L U S I O N SA r t e m i a n a u p l i i c a n b e s t o r e d f o r 2 4 - 4 8 h w i t h o u t l o s in g t h e i r n u t r i -t i o n a l v a l u e f o r f i sh a n d c r u s t a c e a n l a rv a e ; d e n s i t ie s u p t o 8 0 0 0 n a u p l i ip e r m l c a n b e k e p t in m o d e r a t e l y a e r a t e d c y l in d r o - c o n i c a l c o n t a i n e r sa t a t e m p e r a t u r e o f 2 - 4 C .

T h i s i s v a l i d f o r A r t e m i a f r o m M a c a u ( B r a z i l ) , S a n F r a n c i s c o B a y( U S A ) , T i e n t s in ( P e o p l e ' s R e p u b l i c o f C h i n a ) , S h a r k B a y (A u s t r a li a ) ,L a v a l d u c ( F r a n c e ) a n d G r e a t S a lt L a k e ( U t a h , U S A ) , b u t n o t f o rC h a p l i n L a k e ( C a n a d a ) a n d B u e n o s A i r e s ( A r g e n t i n a ) b r i n e s h r i m p th a td i e o f f r a p i d l y d u r i n g s t o r a g e .

T h i s t e c h n i q u e o f c o o l s t o r a g e o f A r t e m i a n a u p l i i o f f e r s u n i q u ea d v a n t a g e s f o r a p p l i c a t i o n i n a q u a c u l t u r e h a t c h e r i e s , e .g . :

1. t h e f r e q u e n c y o f A r t e m i a c y s t - h a t c h i n g a n d n a u p l i i - h a r v e s t i n g c a nb e c o n s i d e r a b l y r e d u c e d ( h a l f t o q u a r t e r o f p r e s e n t a c ti v it ie s );


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7 4 P. LOger, P. Vanhaecke, P. Sorgeloos

T A B L E 4P r o c e n t u a l C o m p o s i t i o n o f F a t t y A c i d M e t h y l E s te r s ( F A M E ) a n d P e rc e n t T o t a lL i p i d ( o n a D r y W e i g h t B a s i s) o f Artemia N a u p l i i f r o m R e f e r e n c e Artemia C y s t s

S t o r e d U n d e r V a r i o u s C o n d i t i o n sFAME Storage conditions

0 h 24 h, 48 h, 24 h, 48 h,2-4C 2-4C 25C 25C

1 4 : 0 1 . 8 6 1 . 8 9 1 . 8 4 1 - 8 9 1 . 5 91 4 : 1 2 . 2 3 2 . 2 3 2 - 2 0 2 .3 3 1 -2 51 5 : 0 0 . 8 3 0 . 8 5 0 . 8 5 0 . 8 7 0 . 7 21 5 : 1 0 - 9 4 0 " 9 4 0 . 9 5 1 . 0 5 0 . 6 51 6 : 0 1 3 . 6 5 1 3 . 5 0 1 3 - 1 6 1 3 - 0 2 1 2 - 0 41 6 : 1 6 0 7 1 6 - 3 9 1 6 - 1 8 1 5 - 8 5 1 5 . 8 9 1 1 . 8 81 6 : 2 6 0 7 o 1 7 : 0 2 . 2 2 2 - 2 8 2 - 1 8 2 . 2 7 1 . 501 6 : 3 6 o 4 - 1 7 : 1 6 0 8 3 . 6 6 3 . 8 9 3 . 8 6 3 - 76 2 . 2 11 8 : 0 3 . 2 4 3 . 2 8 3 - 3 4 3 .9 1 6 . 1 01 8 : 1 6 0 7 /6 0 9 3 1 . 1 9 3 1 . 4 5 3 2 . 3 2 3 1 . 8 0 3 6 - 8 61 8 : 2 6 0 6 9 . 7 8 9 . 4 1 1 0 - 0 0 8 - 9 6 8 . 8 42 0 : 0 }18 3603/606 a 1 -30 1 .17 1 .08 1-30 1-372 0 : 1 6 0 7 / 6 0 9 0 . 9 4 0 - 9 1 0 . 7 9 0 - 9 0 1 .1 11 8 : 4 6 0 3 . . . .2 1 : 0 0 -3 1 0 -3 1 0 . 3 2 0 . 3 3 0 . 1 02 0 : 2 60 6/609 . . . . .2 0 : 3 60 3 0 - 1 6 0 . 0 8 0 . 0 6 0 . 0 5 -2 0 : 4 6 0 3 / 6 0 6 4 . 2 4 4 . 2 8 4 . 2 0 4 . 5 0 5 . 9 92 2 : 1 0 . 0 6 t r a c e - -2 0 : 5 60 3 7 - 0 5 7 . 0 7 7 - 0 7 7 . 2 2 8 . 0 7% T o t a l l i p i d / d r y 2 0 . 9 4 2 0 . 4 7 2 0 - 7 0 b b

w eig ht -+0"82 -+1-06 -+1-15a M o r e t h a n 9 9 % - 1 8 : 3 6 0 3 .b N o d a t a a v a i l a b le .


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In ternat ional s tudy on Artemia - X X I V 75

T A B L E 5Results of the Mysid Culture Test with Ar t em i a Nauplii (Macau no. 871051) Stored

Under Various ConditionsStorage condit ions of Artemia nauplii0 h 2 4 h , 2 - 4 C 4 8 h , 2 - 4 C

Percent survival 89.7 -+ 8-9 96.5 -+ 13.5 93-3 -+ 13-6Individual dry weight (/ag) 254 -+ 28-0a 352-6 -+ 55.3 ~ 335.8 -+ 33"8 bIndividual length (~m) 4 376 -+ 233 4 591 -+ 200 4 593 -+ 220Reproductive characteristics

% Sexual differentiation 100 100 100% di/dd 14.6 6-7 3"7% 9i/99 53-0 33-0 18-3% 9, /9 9 43.6 53-1 78.0% ~/99 3-3 13-9 3-7

~ i , Immature male; 9 i , immature female; 86, total number of males; 99, totalnumber of females; 9, , female with eggs in ovaria; ~, female with eggs in marsupium.a'bMeans with a different superscript (a ,b ) are significantly different (P~< 0-05).

T A B L E 6Results of the Carp Culture Test with Ar t em i a Nauplii (Macau no. 871051) Stored

Under Various ConditionsCarp results Storage con dit ions o f Artemia nauplii

O h 2 4 h , 2 - 4 C 4 8 h , 2 - 4 CPercentage survivalIndividual wet weight (mg)

95.7 -+ 1.4 95-7 + 1.4 93.3 -+ 0.8173 + 2'9a 169.5 -+ 6.4 a 159'3 -+ 3.2 b

a, b Means with different superscript (a,b) are significantly different (P ~< 0.05).


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76 P. L bger, P. V anh aecke, P. Sorgeloos

c

iii(

~m.

~

Fig. 1. Schematic diagram of auto m atic distr ibut ion system for coo led Artemianauplii (m odified from IA ge r and Sorgeloos, 1982 b). (a) Tan k fil led with r insingwa ter; (b) elec trom agne tic valve; (c) refrigerator; (d) stock-cy linder w ith Artemia

nauplii ; (e) on e-w ay valve; (f) peristalt ic pum p; (g) to cul ture tan k.

2 . n a u p l i i d i s t r i b u t i o n f r o m a c o o l e d s t o c k t o t h e p r e y i n g la r v a e c a nb e a u t o m a t e d , e .g . w i t h t h e s y s t e m o u t l i n e d i n F ig . 1. T h e f re -q u e n c y o f l iv e - f o o d d i s t r i b u t i o n c a n b e i n c r e a s e d a t w i ll , a s s u ri n gs h o r t e r r e t e n t i o n t im e s o f t h e Artemia n a u p l i i i n t h e c u l t u r e t a n k st h u s p r o v i d i n g a h i g h e r q u a l i t y f o o d f o r t h e p r e y i n g l a r v a e ;

3. Artemia l e f t o v e r s a r e n o t w a s t e d b u t c a n b e s to r e d f o r la t e r us e .


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I n t e rna t i ona l s t udy on A r t e m i a - X X I V 77A C K N O W L E D G M E N T S

W e a re v e r y i n d e b t e d t o D r E . J a s p e r s fo r r e a d in g t h r o u g h t h e m a n u -s c r i p t .

R E F E R E N C E SBeni j t s , F . , Vanvoorden, E. & Sorge loos, P . (1976) . Changes in the b iochemica l

composi t ion of the ear ly la rva l s tages of the br ine shr imp, Artemia sa l ina L. In :Proc . l O th European Sum pos i um on M ar i ne B i o l ogy - 1 , eds G. Persoone & EJaspers, Universa Press, Wetteren, pp. 1-9.

JDy e , J . E . (1980) . The p rodu c t ion an d e f f i c i en t u se o f f r e sh ly -ha tched b r ine sh r impnaupl i i A r t e m i a in the larval rearing of m arine f i sh a t the ha tcher ies o f theBri t i sh Whi te Fish Author i ty . In : The Br i ne Shr i m p A r t e m i a - 3 , e d s G . P e rs o on e .P. Sorgeloos, O. Roels & E. Jaspers, Universa Press, Wetteren, pp. 271-6.

Hen t sche l , E . (1968) . D ie pos tem bryo na len E n twick lungss t ad ien yon Artemia sa l inaLeach be i ve r sch iedenen Tem pera tu ren (A nos t raca , C rus tacea ) . Z o o l . A n z . , 180,3 7 2 - 8 4 .

Kin ne , O. (1977) . Cul t iva t ion of animals - - research cul t iva t ion . In : M ar i ne Eco l ogy ,vol. 3 , par t 2 , ed . O. Kinne , Jo hn Wiley & Sons, Chicheste r , pp . 5 79-1 293 .

l_~ger , Ph . & Sorge loos, P . (19 82a ) . Descr ip t ion o f a s tand ard cul ture tes t w i thjuven i l e s o f the mar ine mys id Mysidops i s bahia Molenock fo r the nu t r i t i ona leva lua t ion of l ive A r t e m i a naupl i i . ( In prepara t ion . )

l_~ger , Ph . & Sorge loos, P . (198 2b) . Au tom at ion in s toc k-cul ture m ainten ance andjuven i l e sepa ra t ion o f the mys id Mysidops i s bahia Molenock . Aquacu l t u ra tEngineer ing , 1 , 4 5 - 5 3 .

M addox , M. B. & Manzi, J . J . (1976 ) . The e ffec ts of a lga l supplem ents on s ta t icsys t em cu l tu re o f Macrobrachium rosenbergi i De Man larvae. In: Proc. 7thAn nu a l M ee t i ng o f t he W orl d M ar i cu l tu re Soc i e t y , ed. J . W. Avau l t , J r . , Louis ianaS ta t e Un ive rs i ty , Ba ton Rouge , pp . 677 -98 .

Mil le r, D. C . , L ang, W. H. , Marcy, M. , Clem, P. & Pechenik , J . (1979 ) . In te rn a t iona ls t u d y o n A r t e m i a . Naup l i a r l ocomoto ry ra t e s , pa t t e rns and pho to re sponses : Acompara t ive s tudy o f va r ious s t r a ins of Ar temia sa l ina . In : B o o k o f A b s t r a c t s ,I n t e rn a t i o n a l S y m p o s i u m o n t h e B r in e S h r im p , Artemia salina, Corpus Christ i ,Texas, U SA, 20 - 23 Augus t , 1979 . Ar t em i a Refe rence Cen te r , Gh en t , p . 91 .

Morr is , R . W. (1956) . Some aspec ts of the problem of rear ing marine f i shes . Bull .Ins t . Oceanogr . Monaco, 1082, 1-61.

Persoone , G. & Sorge loos, P . (1972) . An improved separa tor box for A r t e m i anaup l i i and o the r pho to tac t i c inve r t eb ra t e s . Helgoldnd er w iss . Mee resunters . ,2 3 , 2 4 3 - 7 .


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78 P. Lkger, P. Vanhaecke, P. SorgeloosSchaue r , P . S . & S i m ps on , K . L . ( 197 8) . E f f ec t s o f d i e t s on t he f a t t y ac i d com -

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