Proc. World Maricul. SOC. 10:781-787 (1979)

S T U D I E S ON COMPOUNDED D I E T S

FOR P e n a e u s k e r a t h u r u s S H R I M P

Fam6n Ferngndez and Francisco Puchal Dpto. de Biologia y Bioquimica

Universidad P o l i t & n i c a de Barcelona Urge1 187, Barcelona 36, Spain

ABSTRACT

The apparent l a r g e d i f f e r e n c e s i n n u t r i t i v e requirements reported f o r d i f f e r e n t shrimp s p e c i e s precludes e x t r a p o l a t i o n o f a v a i l a b l e d a t a from one s p e c i e s t o another . This f a c t and t h e genera l s c a r c i t y of knowledge about c rus tacean d i e t a r y requirements p r e s e n t s a formidable problem t o t h e n u t r i t i o n i s t when beginning with a new shrimp candidate .

The s p e c i e s used i n t h e s e experiments w a s t h e Penaeus kera thurus , a shrimp n a t i v e t o t h e e a s t e r n A t l a n t i c and Mediterranean c o a s t l i n e . A t o t a l o f 1,000 p o s t l a r v a e were used i n s i x experimental t r i a l s , concerned w i t h t h e a c c e p t a b i l i t y of formulated r a t i o n s and t h e e f f e c t i v e n e s s of d i f f e r e n t p r o t e i n sources , and cons ider ing previous techniques c a r r i e d o u t i n t h i s f i e l d . Other aspec ts examined w e r e t h e inf luence of n a t u r a l food (mussel mant le and gonad) as a complement t o t h e a r t i f i c i a l d i e t , and t h e presence i n it of an u n i d e n t i f i e d growth f a c t o r ( s ) .

From t h e r e s u l t s obtained, t h e a c c e p t a b i l i t y of t h e compounded d i e t s s t a n d s out . However, t h e s e d i e t s f a i l e d t o suppor t g r e a t e r than 50-55% of t h e f r e s h mussel d i e t growth i n a l l experiments. This observat ion and t h e s t rong reduct ion i n n u t r i t i o n a l va lue of t h e f r e s h food by pro- cess ing a r e r e l a t e d by t w o p o s s i b l e explana t ions : f i r s t , t h e rap id leaching o f water-soluble n u t r i e n t s , and second, t h e idea of a l a b i l e unknown metabol i te e s s e n t i a l f o r the growth of t h e s e animals.

INTRODUCTION

The shrimp Penaeus kera thurus i s a commercially va luable spec ies of t h e Eastern A t l a n t i c and Mediterranean c o a s t l i n e . In t h e n a t u r a l envi- ronment, t h e s e l e c t i o n of food dur ing the year by t h i s spec ies has been s t u d i e d by San F e l i u and Alcaraz (1971). According to these au thors , 80% of t h e stomach content i s c o n s t i t u t e d o f c rus taceans , molluscs and polychetes , i n t h a t o rder . ments, such a s mud and sand. Nothing i s known, however, about s p e c i f i c n u t r i e n t requirements of t h i s s p e c i e s under c o n t r o l l e d c u l t u r e condi- t i o n s .

There i s also an apprec iab le amount of sedi-

During t h e l a s t few y e a r s , a number o f i n v e s t i g a t o r s have been

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involved i n shrimp n u t r i t i o n a l s tud ie s and considerable l i t e r a t u r e i s now ava i lab le on this subjec t . s u l t s obtained with the va r i e ty of shrimp species under d i f f e r ing ex- perimental conditions, together with the apparent l a rge d i f fe rences i n t h e i r n u t r i t i v e requirements, make it d i f f i c u l t i s not impossible to ex- t r apo la t e ava i lab le da ta from one species t o another.

The ob jec t of t h i s work was, therefore , t o test the acceptab i l i ty of formulated r a t ions and the e f fec t iveness of d i f f e r e n t pro te in sources, considering previous techniques ca r r i ed o u t i n t h i s f i e l d , i n order t o f i n d a standard d i e t which would replace t h e f resh mussel mantle and gonad t i s s u e present ly used a s a cont ro l .

Nevertheless, t h e g r e a t d ive r s i ty of re-

MATERIAL AND METHODS

A t o t a l o f 1,000 post-larval shrimp hatched i n the Fishery Research I n s t i t u t e , Castellon (Spain), were used i n t h e severa l experimental t r i a l s reported i n t h i s paper. The shrimp were lodged i n rectangular flow-through seawater tanks (300 l i t e r s and 0.5 m 2 ) . Each tank w a s pro- vided with a s e t of movable p l a s t i c mesh d iv iders , which allowed varia- t ions i n t h e number of ava i l ab le experimental sections. The v a r i a b i l i t y i n t h e growth pa t t e rn found i n these animals w a s compensated f o r by rep- l i c a t i n g each treatment from 2 t o 4 times.

The experiments were ca r r i ed o u t sequent ia l ly once the conclusions of the preceding ones were known. Since no d i e t was estimated t o be s u f f i c i e n t f o r t he feeding t r i a l s a s t o the n u t r i t i o n a l requirements of t he shrimp, severa l d i s t i n c t l y d i f f e r e n t d i e t s were tes ted .

I n i t i a l l y , t he d i e t s w e r e formulated by l i n e a r programming accord- ing t o some o f the premises pointed out by Kitabayashi e t a l . (1971a-d) and Deshimaru and Shigeno (1972), bu t using typ ica l animal feed ingre- d ien ts . As t he t r a i l s progressed, these d i e t s were replaced by o thers with pro te in of superior b io logica l value, i . e . , squid meal, shrimp meal and freeze-dried mussel mantle and gonad meal. Tables 1 and 2 show the composition and ana lys i s of two of t h e d i e t s which supported the bes t growth.

Agar was chosen a s the binding agent i n t h e preparation of the elaborated d i e t s . A volumetric f l a sk containing seawater with agar was allowed t o stand f o r 15 min i n a ho t water bath a t 100°C, then the f l a sk was heated d i r e c t l y with occasional s t i r r i n g u n t i l t h e agar was com- p l e t e ly melted. After cooling t o 50°C, t he so lu t ion was poured i n t o a p l a s t i c box containing the powdered d i e t and thoroughly mixed. l i d i f i e d t h e moist d i e t s were s tored under r e f r ige ra t ion u n t i l used. For comparative purposes and owing t o the v a r i a b i l i t y i n t h e experimen- t a l and environmental conditions among experiments, na tura l food ( f resh mussel mantle and gonad) was used a s a cont ro l i n a l l cases.

feed allowances of each group t o consumption. Leaching r a t e s were as- sessed by weighing t o t a l dry matter recovered by f i l t r a t i o n a f t e r keep- ing the d i e t 8 hours i n seawater, and the r e su l t i ng f igu res were consid- ered i n t h e ca lcu la t ions of feed consumption.

Once so-

The shrimp wexe fed before sunset t o approximate s a t i e t y , adjusting

TABLE 1. Composition o f t h e Experimental D i e t s i n Percent

Ingredien ts D i e t 1 D i e t 2

Squid meal 16.0 50.0 Shrimp m e a l 30.0 16.0 F ish meal--65% p r o t e i n 8.0 8.0 Molasses yeast--70% p r o t e i n 7 .O 7.0 Dextr in 18.5 8.5 Wheat f l o u r 10.0 0.0 Calcium carbonate 1.0 1.0 Calcium phosphate 1.0 1.0 Glucosamine--HC1 0.5 0.5 Glucose 3.5 3.5 Lec i th in 1.0 1.0 Choles te ro l 0 .5 0.5 Vitamin mixa 2 .o 2.0 Mineral mixb 1.0 1.0

Percent t o t a l

Seawater, g/100 q Agar, d 1 0 0 9

100.0 100.0 3.5 3.5

200.0 200.0

aVitamin mix p e r kg o f d r y d i e t : vi tamin A, 25,000 I U ; vitamin D, 2,400 I U ; v i tamin K, 20 mg; vi tamin E , 100 mq; r i b o f l a v i n , 40 mg; nia- c i n , 200 mg; Ca-pantothenate, 300 mg; pyridoxine (HCl), 60 mg; f o l i c a c i d , 4 mg; c h o l i n e c h l o r i d e , 3000 mg; cobalamine, 0.4 mg; b i o t i n , 2 mg; i n o s i t o l , 2000 mg; ascorb ic a c i d , 500 mq; thiamin ( H C l ) , 20 mg; ethoxy- quin, 300 mg.

bMineral mix p e r kg of dry d i e t : i r o n , 18 mg; z i n c , 24 mg; copper, 4 mq; i o d i n e , 1 mg; manganese, 44 mg; magnesium, 80 mg; c o b a l t , 0.2 mg.

TABLE 2 . Analysis of the Experimental D i e t s and Mussel Mantle

Parameters D i e t s ( % on dry b a s i s ) 1 2 3a

Pro te in 40.42 59.38 64.70 Fiber 3.07 1.80 - F a t 4.64 7.76 13.75 Ash 9.12 7 .43 7.62 C a l c i u m 2.78 2.22 0.22 Phosphorus 0.94 0.80 0.96

Amino Acids ( % o f crude p r o t e i n ) Arqinine 3.29 4.44 5.59 Lysine 3.26 4.74 8.38 Methionine 1.17 1.71 2.37 Threonine 1.84 2.97 5.16 Methionine+Cystine 1.45 2.13 3.48

aMussel mantle and gonad.

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RESULTS

The t y p i c a l p a t t e r n of r e s u l t s comparing t h e f r e s h food c o n t r o l and d i e t s 1 and 2 is shown i n Table 3. S imi la r t o o t h e r experiments, t h e r e w e r e no s i g n i f i c a n t d i f f e r e n c e s (p 0.05) between e labora ted d i e t s ; how- ever , t h e d i f f e r e n c e s between t h e s e d i e t s and t h e mussel i n growth incre- ment and conversion r a t i o were very s t r i k i n g .

TABLE 3 . Experimental Resul t s f o r Penaeus kera thurus Fed on T e s t Diets (Experimental condi t ions : feeding t r i a l , 30 days; temperature, 2321'C; s tocking r a t e , 30 animals/tank; 2 r e p l i c a t e s ; i n i t i a l mean body weight, 3.15 g ; running seawater , 1.75 l i t e r s / m i n / tank)

Treatments 1 2 3a

Surv iva l r a t e ( 9 ) 92 92 95 Growth increment (g ) 0.821 0.887 1.714 Feed consumptionb (g) 0.247 0.250 0.195 Conversion r a t i o c 9.14 8.49 3.41

aMussel mantle and gonad. bAnimals/day. 'Dry food consumed/live weight gained.

DISCUSSION

F i r s t of a l l , t h e s e experiments provide evidence t h a t t h e s p e c i e s Penaeus kera thurus i n g e s t s compounded d i e t s i n a q u a n t i t y s u p e r i o r even t o t h e n a t u r a l food o f mussel mantle and gonad with good s u r v i v a l and without p a l a t a b i l i t y problems. However, the s t rong d i f f e r e n c e s i n growth increment between f r e s h mussel and e labora ted d i e t s , i n a l l t h e experi- ments, r e q u i r e p a r t i c u l a r a t t e n t i o n .

The requirement by penaeid shrimp f o r d i e t a r y s t e r o l , owing t o t h e lack of b iosynthes is of t h i s substance from a c e t a t e , has been repor ted by s e v e r a l au thors (Kanazawa e t a l . , 1971; Shudo e t a l . , 1971; Deshimaru and Kuroki, 1974) . The i n c l u s i o n o f c h o l e s t e r o l i n our d i e t s a t t h e l e v e l of 0.1% r e s u l t e d i n improving t h e growth increment from 30 t o 40% i n r e l a t i o n t o mussel mantle and gonad growth. Fur ther increases of s t e r o l component t o 0.5% i n subsequent d i e t e v a l u a t i o n s , however, d i d n o t show b e t t e r r e s u l t s .

The u t i l i z a t i o n o f p r o t e i n - r i c h meals with s i m i l a r amino a c i d com- p o s i t i o n t o t h e shrimp, i . e . , squid m e a l (Kitabayashi e t a l . , 1971a; Deshimaru and Shigeno, 1972; K i t t a k a , 1975) and shrimp meal (Grajcer and Neal, 1972; Zein-Eldin and Meyers, 1973) produced a not iceable increase i n consumption, b u t f a i l e d t o promote growth rates g r e a t e r than 55% of those achieved using t h e mussel d i e t . S i m i l a r l y , t h e use of compounded d i e t s e labora ted with t y p i c a l animal feed i n g r e d i e n t s and formulated by l i n e a r programming t o match t h e amino a c i d d i s t r i b u t i o n r a t i o of P . ke- r a t h u r u s meat showed t h e same r e s u l t s .

Considering t h e d i v e r s i t y of i n g r e d i e n t s used i n d i e t e l a b o r a t i o n s t o g e t h e r with t h e g r e a t v a r i a t i o n o f n u t r i t i v e parameters r e f l e c t e d i n

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them, it i s d i f f i c u l t t o suppor t t h e idea t h a t i n some ins tances t h e n u t r i t i v e requirements have n o t been s a t i s f i e d . But t h e above r e s u l t s and t h e uniform growth o f j u v e n i l e shrimp reared on experimental d i e t s would sugges t t h e shrimp w e r e n o t rece iv ing some unknown metabol i te(s1 o r s p e c i f i c n u t r i e n t ( s ) e s s e n t i a l f o r t h e growth of these animals. This n u t r i e n t , absent i n t h e e labora ted d i e t s b u t p r e s e n t i n t h e f r e s h mus- se l , would prevent t h e shrimp from expressing t h e i r growth p o t e n t i a l , thus masking t h e response t o t h e d i f f e r e n t d i e t s .

The evidence f o r a growth f a c t o r i n some f r e s h food (mussel mantle and c r a b gonad and d i g e s t i v e gland) was suggested by F o r s t e r and Beard (1973) during t h e i r experiments wi th Palaemon s e r r a t u s . These au thors , s u b s t i t u t i n g t h e compounded d i e t one day a week f o r some of t h e f r e s h foods mentioned above, could improve t h e n u t r i t i v e va lue of these combi- na t ions t o t h e l e v e l of t h e f r e s h mussel mantle i t s e l f . The same sub- s t i t u t i o n c a r r i e d o u t with P. kera thurus produced a s i g n i f i c a n t improve- ment i n growth increment and conversion ra t io , b u t d i d not approach t h e r e s u l t o f f r e s h mussel i t s e l f . I n t h i s r e s p e c t , p a r t i a l explanat ion could l i e i n t h e f a c t t h a t t h e shrimp r e j e c t e d t h e compounded d i e t the day a f t e r feeding with mussel mantle and gonad.

t i t y of f r e s h food a s a complement t o t h e a r t i f i c i a l d i e t t o g e t good r e s u l t s was a l s o showed by Ki t taka (1975) . On t h e o t h e r hand, the in- c l u s i o n of these f r e s h foods i n t o t h e feed wi th t h e required processing techniques d i d n o t improve t h e q u a l i t y of t h e d i e t s themselves. q u a l i t y loss with process ing has been noted f o r a v a r i e t y of crustacean spec ies : P. serratus ( F o r s t e r and Beard, 1973); Homarus americanus (Conklin e t a l . , 1978); Carcinus maenas (Adelung and Ponat, 1977) . Sur- p r i s i n g l y , the s t r o n g reduct ion i n t h e n u t r i t i o n a l value of t h e mussel mantle and gonad w a s even more pronounced when using it exc lus ive ly , i n f reeze-dr ied form, recording p r a c t i c a l l y no growth increment.

The necess i ty o f cont inuing t o feed t h e shrimp with a c e r t a i n quan-

This

I t i s p o s s i b l e t o sugges t two explana t ions of t h e above r e s u l t s . F i r s t , t h e r a p i d leaching o f water-soluble substances leads t o t h e sub- optimal a m u n t o f e s s e n t i a l n u t r i e n t s a v a i l a b l e t o t h e animal. In t h i s c a s e , w e can a l s o assume an i n c r e a s e i n t h e leaching of s p e c i f i c n u t r i - e n t s a f t e r processing t h e f r e s h food. Recent exper inents being c a r r i e d o u t a t t h e Bodega Marine Laboratory are i n v e s t i g a t i n g t h i s p o s s i b i l i t y .

The second explana t ion would be t h e suggest ion r a i s e d by F o r s t e r and Beard (1973) t h a t t h e p o s s i b l e growth f a c t o r ( s ) i s destroyed o r de- natured by process ing of any kind. a c i d s i n f reeze-dr ied s t o r e d Myti lus e d u l i s was repor ted by Ranke and Ranke (Adelung and Ponat, 1977) . This process could a l s o a f f e c t o t h e r p a r t i c u l a r components o f t h e f r e s h food s t r o n g l y implicated i n t h e growth of these animals . In a d d i t i o n , we cannot d i s c a r d a combination of these two p o s s i b i l i t i e s .

The d e s t r u c t i o n of p r o t e i n and amino

ACKNOWLEDGMENTS

We would l i k e t o express our a p p r e c i a t i o n t o D r . Douglas E. Conklin f o r h i s a s s i s t a n c e i n prepar ing t h i s manuscr ipt .

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