Nutritional of Requirement of Shellfish

Introduction
Techniques for the artificial propagation and culture of tiger shrimp, Penaeus monodon, have been
established. In conjunction with the high market price and demand, the number of tiger shrimp farms and
the production of the species have increased tremendously in the last few years in various Asian countries.
However, problems still remain to be solved. Water quality is closely related to feed and feeding
management. A less polluting diet which is low in nitrogen and phosphorus has to be developed. Also,
nearly half of the factors that can be attributed to tiger shrimp diseases seem to be directly or indirectly
related to nutritional adequacy of diets. Penaeus monodon is the most important aquaculture shrimp
species in the world. The technology to develop mature spawners, propagate larvae and post larvae, and
grow juveniles to commercial size in relatively high density has been well established. Accordingly,
artificial feeds of different forms and composition have also been developed and widely used in every
phase of culture from larval rearing to broodstock maturation and spawning. However, when we review
our understanding of the nutritional requirements of this species and other marine shrimps, there is
considerable limitation in the existing knowledge which hinders the successful development of
nutritionally complete and cost-effective diets. Most of what is known about the nutritional requirements
of tiger shrimp has been derived from the studies of P. japonicus which is believed to be more
carnivorous than P. monodon. Dietary discrepancy due to species differences is obvious. However, until
more studies are conducted to determine the nutritional requirements of P. monodon, feed formulations
for this rapidly expanding shrimp culture industry would still be based largely on intuition and unknown
premises from secret sources rather than nutritional science.

Protein
It has been reported that protein is an essential nutrient for Shrimp. Protein, which is required for growth
and maintenance, is an expensive component in a diet. Excessive protein levels will increase feed cost and
nitrogenous waste. Therefore, knowledge of the optimum level of protein and the protein-sparing effects
of non-protein nutrients such as lipids or carbohydrates would be effective in reducing feed costs and
water pollution.
Salinity is among the factors known to influence the protein requirement of shrimp. A recent study
comparing the optimum dietary protein level for P. monodon reared in seawater and brackish water gave
an inverse relation between protein level and salinity. The optimum protein level for juvenile P. monodon
raised in full strength seawater (40%) has been studied by Shiau et al. (1991). When the energy content
was increased, the optimum protein level for the shrimp raised in seawater could be effectively lowered to
36% (Shiau and Chou 1991). From previous results, we know that shrimp growth is lower under high
salinity than that under low salinity. Although shrimp culture using seawater will benefit from a reduced
protein requirement, thus a lower feed cost; its overall results will still have to be decided by the balance
between reduced feed cost and decreased shrimp growth, plus other related factors. Manipulation of
dietary protein alone may not improve the growth of shrimp raised in seawater. Increased
supplementation of lipid may be helpful in supplying metabolic energy to enhance growth.
Despite much discussion on protein requirement or optimum dietary protein level for shrimp or other
animals, there is one basic concept in nutrition that can not be neglected: animals have requirements for
amino acids rather than proteins. Unless we know the exact requirements for indispensable amino acids,
trying to manipulate feedstuff proteins to achieve an efficient diet will be most likely fruitless. While
relatively abundant knowledge concerning protein nutrition are available for P. monodon and other
shrimp, there is very little data available concerning amino acid nutrition for successful commercial feed
formulation.

Amino acid requirements

Substitution of high quality proteins with low quality proteins supplemented with crystalline amino acid
does not enhance the growth of some fish and crustaceans including penaeid shrimp. The incorporation of
arginine originating from dietary protein into muscle protein of P. japonicus can be as high as 90.5%,
whereas the assimilation of crystalline arginine into muscle protein is only 0.6% after six days of feeding.
More than 50% of the arginine in an amino acid test diet (all amino acids in the diet were from a mixture
of crystalline amino acids) was absorbed in the posterior part of the alimentary tract of P. japonicus.
These results indicate that P. japonicus and probably other species of shrimp can effectively absorb
crystalline amino acids from the alimentary tract, but do not effectively utilize them in muscle protein
synthesis.
Among the possible explanations suggested for the failure of crystalline amino acids to enhance growth is
the comparatively low efficiency of utilization of free amino acids (Cowey and Sargent 1979). It was
hypothesized that crystalline amino acids used to supplement a protein are assimilated much more rapidly
than those linked by peptide bonds in the dietary protein (protein-bound amino acids) (Cowey and Sargent
1979). Tissue concentration of these supplemental amino acids may thus be temporarily elevated and may
be immediately catabolized rather than used for protein synthesis. If the hypothesis is true, reducing the
rate of release of supplemental amino acids should be advantageous in promoting protein synthesis and
thus growth. In a recent study on juvenile P. monodon, Chen et al. (1992a) demonstrated that L-arginine
microencapsulated with either cellulose acetate phthalate (CAP) or glycerol monostearate (GM) could be
effectively added to the diet of the shrimp. Test diet containing supplementary CAP- or GM-arginine
resulted in a significantly better growth and food conversion than diet supplemented with crystalline
arginine alone. These results indicate that microencapsulation of arginine may have delayed the release of
arginine in the gastro-intestinal tract of the shrimp. The delayed release, and thus absorption, may have
increased the effective provision of arginine to shrimp juveniles. With the use of microencapsulated L-
arginine, the quantitative requirement for the amino acid was determined (Chen et al. 1992b). The
arginine level required for optimal growth, determined by a broken-line analysis of weight gain, was 25.0
g/kg diet or 54.7 g/kg protein.

Carbohydrates
Carbohydrate is the most economical dietary energy source (cost/kcal). There is little information on the
carbohydrate nutrition of shrimp. The types and levels of carbohydrate in the diet have been shown to
affect the growth of P. japonicus and P. duorarum. For juvenile P. monodon, a significant influence of
different types and levels of carbohydrate in the diet on survival is observed. P. monodon utilize trehalose
and sucrose better than glucose. Among P. monodon fed glucose, dextrin and starch diet, those fed starch
showed the best weight gain followed by those fed dextrin and glucose (Table 3). Poor utilization of
glucose by P. monodon found in the study is in general agreement with Alava and Pascual (1987).
Furthermore, Shiau (1992) found that as the dietary starch level increased from 20 to 30% and dietary
protein levels decreased from 40 to 30%, protein digestibility and dry matter digestibility were not
affected (P > 0.05) but protein deposition significantly increased (Table 4). In contrast, weight gains were
reduced (P > 0.05) when dextrin and glucose were used as carbohydrate sources (Table 3). It is reasonable
to suggest that the optimal dietary protein level for P. monodon would be lower if starch were chosen as
the carbohydrate source instead of glucose or dextrin. The poor ability to utilize glucose has also been
reported for other penaeid species such as P. japonicus (Kitabayashi et al 1971b; Deshimaru and Yone
1978; Abdel-Rahman et al. 1979), P. setiferus (Andrews et al 1972) and P. duorarum (Sick and Andrews
1973). The mechanism of the poor utilization of glucose is not yet fully understood. Abdel-Rahman et al
(1979) reported that serum glucose levels in P. japonicas increased rapidly after administration of glucose
and remained at high levels for 24 hours. When the diet contained disaccharides, serum glucose increased
to a maximum level at 3 hours and then decreased to a low level. The authors suggested that dietary
glucose was quickly absorbed from the alimentary canal and released into the hemolymph resulting in a
physiologically abnormal elevation of serum glucose concentration thereby impairing its utilization as an
energy source. It was also found that serum glucose levels in P. monodon fed the diets containing glucose
peaked faster than in shrimps fed the dextrin and starch diets.
Another possible factor that may relate to the poor growth of shrimp fed glucose containing diet is the
inhibition of amino acid absorption in the intestine by the presence of glucose (Alvarado and Robinson
1979). Hokazono et al. (1979) reported that the presence of 10 mM glucose reduced the uptake of L-
lysine from 26.64 to 12.34% in the mid-intestine and from 23.24 to 5.4% in the posterior intestine of the
rainbow trout.

Lipids

Lipids refer to a group of fat-soluble compounds found in the tissues of plants and animals and are
classified as fat, phospholipids, sphingomyelin, waxes, and sterols. Lipids are important sources of
metabolic energy. In this respect, dietary lipids may be used to spare the more valuable protein for
growth. In particular, free fatty acids derived from triglycerides are the major energy source for
metabolism in the muscle. Lipids provide essential fatty acids which are essential for the maintenance of
the integrity of cellular membranes required for optimal lipid transport (bound to phospholipids as
emulsifying agents) and are precursors of the prostaglandin hormones. They are also a source of essential
steroids, which in turn perform a wide range of important biological functions. From a viewpoint of feed
technology, lipids act as lubricants for the passage of feed through pellet die, as substances which reduce
the dustiness of feeds, and as enhancer of feed palatability. Factors such as salinity, temperature, dietary
lipids, and sexual maturity affect the fatty acid composition of fresh fish or shrimp. The fatty acid patterns
among crustaceans in freshwater and marine environments tend to be similar to those of fish. Linoleic
acid (18:2n-6) and n-6 polyunsaturated fatty acids tend to be the predominant fatty acids in freshwater
shrimp while linolenic acid (18:3n-3) and 20 and 22 carbons, n-3 highly unsaturated fatty acids
predominate in marine crustaceans. Crustacean lipids exhibit a similar response to changes in
environmental temperature. Both desaturation and chain elongation increase with a decrease in
temperature. The influence of dietary lipids on fatty acid composition of P. setiferus fed a diet high in
18:2n-6 and low in 20 and 22 carbon HUFA was apparent after one month. Specific retention of n-6 fatty
acids was evident. The n-6/n-3 ratio changed from 1:1.8 to 1.6:1 after feeding shrimp diet with n-6/n-3
ratio of 4.5:1. Hayashi (1976) reported higher concentration of n-3 fatty acids in the ovary than in the
body of adult crustaceans. These indicate the trend of dietary n-3 HUFA requirement for marine shrimp
and for broodstock.

Lipid requirement
A study showed that growth of P. monodon was significantly enhanced by cod liver oil at 12%, compared
with 4% and 8%. At the same dietary levels tested, soybean oil and corn oil were good sources of lipid
next to cod liver oil, while coconut oil, pork lard, and beef tallow were poor lipid sources (Catacutan
1991).

Sheen and Chen (1992) reported that P. monodon fed a diet containing 8, 10, or 12% cod liver oil/corn oil
(2:1 w/w) mixture had significantly higher weight gain than those fed lower lipid levels. This shows that
juvenile tiger shrimp do not require an absolute level of dietary lipid. Rather, the optimum amount of
dietary lipid depends on the amount and quality of dietary protein, the level of available dietary energy,
and satisfaction of specific fatty acid requirement. Millamena et al. (1986) found that cod liver oil was a
better lipid source for the broodstock than soybean lecithin or a combination of cod liver oil and soybean
lecithin at 1:1 ratio.

Essential fatty acid requirement

Kanazawa et al. (1979) have shown the absence of de novo synthesis of linoleic (18:2n-6), linolenic
(18:3n-3), eicosapentaenoic (20:5n-3) and docosahexaenoic (22:6n-3) acids from 14C acetate or 14C
palmitic acid in P. monodon. This suggests that P. monodon require some of these fatty acids as essential
nutrients. Catacutan (1991) reported that 2.6% dietary n-3 HUFA enhanced growth of juvenile P.
monodon, but levels of 18:2n-6 higher than 5% have a negative effect. Kontara (1986) found that P.
monodon larvae which received Artemia enriched with n-3 HUFA had higher growth and survival rate
than shrimp larvae fed Artemia nauplii of the same strain. Millamena (1989) showed that the fatty acid
composition of broodstock diet influenced the tissue fatty acid patterns and hatchability of eggs from
pond-reared P. monodon. Females fed a diet high in 20:4n-6, 20:5n-3, and 22:6n-3 highly unsaturated
fatty acid and n-3/n-6 fatty acid ratio gave higher percentages of fertilized eggs and higher hatching rate
than females fed diets with high phospholipid and lower n-3 HUFA. Catacutan (1991) and Millamena
(1989) indicated that n-3 HUFA was an essential fatty acid for P. monodon and the requirement was
estimated (from 15.03% n-3 HUFA in a 12.14% dietary lipid) to be 1.8-2.6% of the diet.
Carotenoid
Shrimp, like other crustaceans, entirely depend on their dietary supply of carotenoid. The major
carotenoid found in crustacean tissues and responsible for typical color of P. monodon is astaxanthin
(Latscha 1989; Boonyaratpalin et al. unpubl.; Howell and Matthews 1991). Nutritional deficiency with
respect to carotenoids has been suggested as the cause of blue disease in farmed P. monodon Although P.
monodon can convert canthaxanthin to asthaxanthin, the efficiency of conversion was only 2-3 to 1 or
lower (Boonyaratpalin et al., unpubl.). Distinction of color was developed in 15 days of feeding a diet
containing 50-75 ppm astaxanthin.

Vitamin requirements
All of the eight B-complex, water-soluble vitamins are known to be required by fish although the
requirement for each has not been established for every cultured species (NRC 1981,1983). For P.
japonicus, the suggested dietary requirement for thiamin is 60 mg/kg and pyridoxine, 120 mg/kg of diet
(Deshimaru and Kuroki 1979). Requirements were estimated on the basis of differences in growth and a
decline in whole animal tissue levels of the vitamin. Deficiencies of thiamin and pyridoxine in fish are
often characterized by nervous disorders. Over a 12-week period, the growth rate of juvenile shrimp
reared on the thiamin-deficient diet was only slightly reduced and survival was similar to those groups
reared on the supplemented control diet. Total body thiamin content in shrimp reared on the deficient diet
was 0.4 mg/kg. Shrimp provided with 60 mg thiamin/kg diet had a total body content of 1.0 mg
thiamin/kg; tissues became saturated at 1.4 mg thiamin/kg as feed levels were increased to above 120
mg/kg diet.
Lack of pyridoxine led to a decrease in both survival and growth. Tissue levels of shrimp grown on the
deficient diet after 12 weeks was only 0.08 mg pyridoxine/kg of whole animal. Survival and growth were
best at a pyridoxine level of 120 mg/kg feed. At this level, the vitamin content of the shrimp increased to
0.43 mg/kg shrimp. Doubling the dietary pyridoxine level saturated the total body level at 0.54 mg/kg, but
growth did not improve. In a 15-week feeding trial, juvenile P. monodon (initial mean weight, 0.13 ± 0.05
g) fed purified diets containing six levels (0, 12, 16, 20, 40, and 80 mg/kg diet) of supplemental riboflavin
showed no significant differences in weight gains, feed conversion ratios, and survival. The dietary
riboflavin required for optimum growth in P. monodon was found to be 20.3 mg/kg diet based on the
broken-line analysis of body riboflavin data. Shrimp fed non supplemented diet did not show apparent
deficiency signs, except for reduced ratios of shrimp head length to body length (Chen and Hwang 1992).
Kanazawa et al. (1976) reported that the provision of choline chloride at 600 mg/kg clearly improved
growth and survival of P. japonicus juveniles. Many vertebrates which can synthesize choline require an
additional dietary source during periods of rapid growth and when fed diets high in lipids. Inclusion of
inositol at 2,000 mg/kg of diet maximized both growth and survival of P. japonicas (Kanazawa et al.
1976). One uncertainty with regard to inositol is its availability. Plant materials, particularly seeds,
contain a high level of inositol but it tends to be complexed as phytic acid, an indigestible form for non-
ruminant farm animals. The requirements for some vitamins such as ascorbic acid were apparently higher
for P. japonicus larvae than for juveniles. It is conceivable, however, that some vitamins may have
leached into the water before feeding. Knowing the exact requirement for each of B-vitamins would be of
little advantage to the shrimp culturist because the cost of these vitamins is relatively low compared with
the overall cost of the feed. It is economically feasible to continue the present practice of providing
generous amount of supplemental B-vitamins. A potential problem with the above approach is the
tendency to provide excessive amounts (just to be on the safe side). Although conventional wisdom
suggests that toxicity is a concern only in fat-soluble vitamins, there are cases where decreased growth
rates of shrimp and prawns were noted as dietary levels of water-soluble vitamins were raised beyond an
optimum. Kitabayashi et al (1971) reported that an excess of vitamin C inhibited growth of P. japonicus.
Doubling the recommended amount (120 mg/kg) of pyridoxine decreased growth performance of P.
japonicas (Deshimaru and Kuroki 1979).

Minerals
It appears that shrimp can absorb some minerals from the water by drinking and by direct absorption via
the gills, skin or both (Deshimaru et al 1978). Minerals serve as structural components of hard-tissue
matrices (e.g., bone, fin, rays, scales, teeth, and exoskeleton) and components of soft tissues (e.g., sulfur
in proteins, phosphorus in phospholipids, and nucleic acids). They are also components of metalloproteins
(e.g., iron in hemoglobin, copper in hemocyanin, zinc in carboxypeptidase), and serve as cofactors and/or
activators of a variety of enzymes
(e.g., zinc activation of alkaline phosphatase). The more soluble minerals (calcium, phosphorus, sodium,
potassium, and chloride) function in osmoregulation, acid base balance, and production of membrane
potentials. With the exception of osmoregulation, the maintenance of osmotic balance between body
fluids and the water in which the animals live, the biochemical functions of minerals in aquatic species
appear to be similar to those in terrestrial animals (Lovell 1989).

Calcium and phosphorus

The calcium requirement may be totally or partially met through absorption of calcium from the water
(Deshimaru and Yone 1978). The low concentration of phosphorus in freshwater and seawater makes a
dietary source of phosphorus potentially more critical. Dietary phosphorus requirement of 1%
(Kitabayashi et al.
1971a), 1-2% (Kanazawa et al. 1984), and 2% (Deshimaru et al. 1978) have been recommended for P.
japonicus. A dietary Ca: P ratio of 1:1 with 1% dietary Ca and
1% dietary phosphorus has been recommended for P. monodon (Bautista and Baticados 1989). It appears
that calcium affects phosphorus availability and calcium levels in excess of 2.5 should be avoided. In
general, bioavailability of minerals especially phosphorus has been found to be positively correlated with
the solubility of the mineral in water. Monobasic phosphates of sodium or potassium are highly available
(90-95%) sources of phosphorus for all species of fish and shrimp (Lovell 1978; Ogino et al. 1979;
Sakamoto and Yone 1979). Availability of the dibasic and tribasic calcium phosphate differs from species
to species. For carp which lacks an acidic stomach, tribasic and dibasic calcium phosphate have an
availability of only 13% and 46%, respectively. The optimum pH in shrimp stomach is 7.0-8.0, which
suggests that utilization of di- and tricalcium phosphate is likely to be poor. Care should be taken to
minimize the over supplementation of dietary phosphorus. Considering that there is a great concern on the
deleterious effects of phosphorus from effluents of aquaculture facilities, minimizing phosphorus
supplements would not only be economical but also a practical approach in reducing phosphorus
pollution.
Magnesium and potassium
Deshimaru and Yone (1978) found that supplementation of 0.3% magnesium did not improve the
nutritive value of semi-purified diet for P. japonicus. Kanazawa et al. (1984) re-evaluated the magnesium
requirement of P. japonicus and reported that dietary supplementation of 0.1-0.5% magnesium improved
the nutritive value of the diet. Davis (1990), utilizing semi-purified diets, found that the individual
deletion of potassium did not result in a significant depression in tissue potassium or growth of P.
vannamei. However, tissue level of magnesium was affected. This indicates a potential interaction
between potassium and magnesium which has to be evaluated.
Kanazawa et al. (1984) reported that diets containing 0.9% potassium improved growth of P. japonicus
compared with diet containing 1.8% potassium.