UNDP/FAO PROJECT THA/89/003
FISH NUTRITION AND AQUACULTURE DIETS

By

John E. Halver

HALVER CORPORATION

Seattle, Washington, USA

Hyperlinks to non-FAO Internet sites do not imply any official endorsement of or responsibility for the opinions, ideas, data or products presented at these locations, or guarantee the validity of the information provided. The sole purpose of links to non-FAO sites is to indicate further information available on related topics.

This electronic document has been scanned using optical character recognition (OCR) software. FAO declines all responsibility for any discrepancies that may exist between the present document and its original printed version.

TABLE OF CONTENTS

Terms of Reference
Review of Progress on THA/87/004
Amino Acid and Protein Study Programme
Lipid Research Programme at NIFI
In-Service Training
Nutrition Laboratory and Diet Development Needs
General Nutrition Consultancy
Recommendations
Personnel Contacted

NUTRITION CONSULTANCY REPORT 6/7 - 26/8/90 - HALVER

TERMS OF REFERENCE POST 11-03:

Description of Duties:

1. To formulate and initiate a research programme to develop optimal needs for freshwater prawn and marine shrimp including:
   1. demonstration of lipid and amino acid analysis techniques;
   2. designing of nutritional experiments for brood stock and grow-out;

   3. advise on feed formulation for freshwater prawn and marine shrimp;

2. To provide in-service training of national staff on shrimp nutrition with special reference to amino acids and lipids.

3. To advise on the appropriate equipment needed for studies on lipids and amino acids, and their installation and operation.

4. To submit a technical report at the end of his assignment.

On arrival the TOR were focused upon demonstration of fatty acid profile analysis and development of the amino acid and protein requirement studies programme at NIFI. Amino acid analysis on the HPLC equipment was defered to a later period since the fume hood was not yet installed and preparation of the phenyisocyanate derivatives of the amino acids was too dangerous without a proper fume hood. A demonstration of reference amino acid standards derivatives on hand was performed for the in-service trainees in a special week long intensive nutrition review requested by the Government Project Coordinator. Protein requirement studies for Pangasius catfish were initiated. Special emphasis was also placed upon developing the capability and training personnel to determine and monitor vitamin C levels in diets and fish tissue. The fume hood listed as a primary need in the THA/87/004 consultancy report submitted in July 1989 was finally installed and became operational only during the last two weeks of this consultancy period. The experimental feed mill has not been completed and therefore only research planning and research proposal development was possible for diet development, diet manufacture and diet testing experiments for shrimp and catfish.

1. Review of Progress on THA/87/004

1.1 Dr. Mali Boonyaratpalin completed the study initiated last year with Sea Bass fed PHOSPHATAN-C (Vitamin C₃) as the vitamin C source in moist diets. All levels tested failed to provide adequate vitamin C intake and fish become scorbutic. In addition the donated supplies of ascorbate polyphosphate received from UTI became discolored upon storage at ambient temperature and could not be used one month after receipt.

1.2 Nanthiya Unprasert completed 50 assays of amino acids in various feedstuffs and diets used for shrimp and fish diets using the HPLC equipment in the new laboratory. She prepared the Phenylisocyanate derivatives in the preparation room without benefit of an adequate hood - - the temporary hood constructed at the station had insufficient air evacuation and therefore she burned her hands and fingers repeatedly during the preparations. I ordered a stop to amino acid derivative preparations for the HPLC analysis until a proper hood was available to protect personnel. Therefore no amino acid analysis was possible during the current consultancy. This situation has now been remedied, albeit only very recently, and Dr. Hardy should now be able to proceed with vitamin preparation for the HPLC analysis programme.

1.3 Vitamin C requirements for Pangasius catfish were initiated using laboratory preparations of semi-moist diets containing coated C which were then dried and stored in the refrigerator until fed. Eighteen lots of fish held in the wet laboratory circular tanks were started on 6 diets 2 July 90 containing 0, 25, 50, 100, 150, 200 mg of coated C per Kg dry diet. Fish have been fed at about 3 % body weight per day. At date of this report no scurvy has been detected in any lot (C stores not yet exhausted). When scurvy appears, repletion studies are planned using samples of L-ascorby1–2-sulfate (ASTON) donated by Pfizer Inc.; L-ascorly-2-triphosphate (STAY-C) donated by UTI, coated C, and reference L-ascorbic acid (C₁). Assays for C₁ content of fish diets and for C₁ and bound C content of fish tissue were performed. Residual C activity in the prepared diets, dried and kept frozen or in the refrigerator was very low indicating most of the bound C was lost during the preparatory or 6 week diet storage period.

1.4 A brief summary of results of the vitamin A, C, E and D experiments was reviewed with Nanthiya Unprasert before her departure for graduate study at Mississippi State University.

1.5 The study with growth promoter added to Penaeus monodon diets indicated about 15 % increase in growth when the promoter was added compared to the same diet devoid of the compound.

1.6 The following recommendations in consultancy report for THA/87/004 of 15 July 1989 were completed: 4.1 the clean room was done, 4.2 an additional air conditioner was installed in clean room; 4.8 a microwave oven was purchased; 4.9 the quality control proximate analysis room was completed including installation of a fume hood; 4.10 the milling equipment budget was increased by US$ 42,000; 4.17 two scientists were sent to the training school at University of Washington.

2. Amino Acid and Protein Study Programme

2.1 Amino Acid requirements of fish and shrimp determine growth rate and protein source profiles in efficient economical formulated feeds. A first approximation of indispensable amino acid requirements may be obtained from amino acid analysis of fish flesh and fish eggs. Therefore a three year amino acid and protein requirement experimental programme was developed with Dr. Wimol Jantrarotai submitting proposals for amino acid analysis using the HPLC equipment for profiles in tissues and eggs, then using this foundation information to develop amino acid increment test diets for experimental feeding of fish, and finally using experimental results to confirm quantitative requirements for the 10 IAA, then formulating practical diets from available feedstuffs for testing under practical rearing conditions. See Annex A.

2.2 Amino acid analysis of fish tissues and fish eggs is planned as soon as Dr. Hardy arrives on about 1 October 90. The Government Project Coordinator urged that catfish be examined first to be followed later by shrimp tissues and eggs. Dr. Wimol has been briefed on operation of the HPLC for amino acid analysis and Dr. Hardy is experienced in amino acid assays as well as vitamin assays. Therefore this element of the amino acid and protein study programme should be implemented early in the next nutrition consultancy period.

2.3 Test diets for determining amino acid quantitative requirements were planned using the Mertz - Halver technique of feeding a test diet containing whole protein components supplemented with crystalline amino acids to duplicate whole egg amino acid profiles except for graded levels of the indispensable amino acid to be tested. Estimates of material needed for these studies were made and purified test diet ingredients were ordered. These experiments will be initialed as soon as the amino acid profiles of flesh and eggs are completed. (See schedule in Annex A).

3. Lipid Research Programme at NIFI

3.1 Fatty acid analysis capability was demonstrated to Dr. Wimol Jantrarotai, Pairat Kosutarak, and Amornrat Sermwatanakul by Nanthiya Unprasert. Folch extraction of liver tissue from catfish, from diets, and from brine shrimps were made by the trainees following the techniques outlined in Watanabe's laboratory manual. After dried total lipid was obtained, the lipid was saponified, then acidified to yield free fatty acids hydrolysed from the triglycerides and phospholipid present in the tissues. These free fatty acids were then methylated and dried before dilution to levels appropriate for GLC fatty acid analysis. Each trainee then injected his/her sample into the gas chromatograph and estimated extracted lipid. Results were compared with fatty acid profiles of Clarias liver oil, pollack liver oil and fatty acid methyl ester standards.

3.2 Amornrat Sermwatanakul plans to assay for fatty acids in various brine shrimp preparations and larval diet mixtures she is testing during development of a larval feed for fish and shrimp.

3.3 New lipid research personnel are necessary at NIFI or a better cooperative and exchange programme with NICA is needed before more emphasis can be placed on developing a strong lipid research and diet testing effort for freshwater and marine shrimp nutrition.

4. In-Service Training

4.1 In response to a request from GPC an intensive review on principles of nutrition of fish and shrimp was organized (Annex B). A number of fishery officers, scientists and students attended the one week course schedule (Annex C). Principles and practices of nutrition and diet development were reviewed each morning with a tutorial ending these sessions. Demonstrations were organized for each afternoon. Practices of Proximate Analysis were demonstrated by Pairat and Wimol one afternoon on the new equipment in the feed proximate analysis laboratory. The new GLC was demonstrated for methylester analysis of fatty acids in feeds and fish tissues. The HPLC was demonstrated for amino acid analysis of reference standards on hand by Nanthiya Unprasert. The feed milling equipment was demonstrated by Pairat and Prasert Sitasit.
Details of the lectures and demonstrations can be found in the handouts of outlines used (Annex D).

4.2 Special training was organized for Dr. Wimol, Pairat and Amornrat, the UBC graduate student on thesis research with the Department of Fisheries, Fisheries Extension Division. Various methods for partial and total lipid extraction of feedstuffs, feeds and animal tissues were used and each individual prepared his own lipid preparations. Techniques for non - destructive free fatty acid preparations were performed by each person under close supervision. Careful methylation of the fatty acids was performed by each, then total methylesters were dried and quantitated. The qualitative and semi-quantitative fatty acid profiles of these methyl esters was determined on the Shimadzu Gas Chromatograph. SAFETY in preparation and in SAFE operations was stressed during the training. Chromatograms obtained were compared with Clarias liver oil and reference Pollack liver oil obtained from Dr. Watanabe at Tokyo University of Fisheries. Assays were compared with Watanabe's chromatograms run with a micro capillary column and the NIFI glass coil column. New standards of methylesters of the C20:5n-3; C22:6n-3 as well as the C18:3n-3; C18:2n-6; C18:1n-9; C18-0; and C16:0 were ordered to complete common fatty acid references for those important and essential for normal fish/shrimp physiology, growth and general nutrition.

4.3 Special training was organized for Pairat and Wimol for analysis of various forms of vitamers C. A simple method for extraction and assay for L-ascorbic acid in feeds and in tissues was reviewed with each person preparing standard curves and sample extractions. The general modified Roe-Keother method for C₁ using water, micro wave, and 6 % TCA extraction procedures were used (annex E). All feed samples showed heavy interference and a sample clarification modified procedure was developed (Annex F). Assays for total C yielding tissue bound C were tested and results verified the presence of bound C in animal tissues. Dr. Wimol will work with Dr. Hardy on the HPLC equipment for more exact quantitative determinations using the Felton-Halver method with tandem C-18 Bondapak columns (Annex G).

5. Nutrition Laboratory and Diet Development Needs.

5.1 Laboratory equipment arrangements in the clean room are adequate except for the electrical supply. Inadequate outlets necessitate use of extension cords and multiple outlets. Only one outlet is properly grounded to protect the expensive equipment, therefore a new set of outlets is needed above the work benches. In addition more outlets are needed on the wall above the outer room work tables. All these should be grounded or at least each outlet should have a + - indication on the main and ground side.

5.2 Outer preparatory table space is inadequate and can be easily improved by installing a work table under the observation windows into the clean room. Electric outlets are also needed above this work table.

5.3 Safety chains are needed for the gas cylinders in the work area by the hood and by the entrance door.

5.4 The proximate analysis and feed quality control laboratory needs an improved “adequate” water supply for the fiber extracting machine. Low water pressure prevent use for fiber determinations which are very important in assessing the nutrients present and cost effectiveness of diet formulations.

5.5 A list of minor equipment to facilitate assays in the nutrition laboratory is listed in Annex H.

5.6 The feed milling equipment is not organized into an efficient line operation. Several pieces of equipment have not been ordered; VIZ; boiler, hammermill, mixer, dryer, conveyors, etc. These were planned in July 1989 but orders and requisitions have been postponed pending receipt of quotations from local and regional equipment suppliers. It may be doubtful if equipment ordered now can be delivered and installed as a complete heat extruded pellet mill for shrimp and fish feeds in time to manufacture diets for field testing which is one major objective of Project THA/89/003, the modified project implemented in January, then June 1990. The existing CPM compressed pellet mill is still operational but one mill is down because of lack of appropriate Die for pellet manufacture. When the new boiler is ordered and installed, and a new die is purchased, this old but operational unit will expand capabilities for experimented diet formulation and pelleting for practical feed treats. These extensive delay may jeopardize the completion of the diet milling capabilities of NIFI and relegate the field diet testing to some future date beyond the tenure of THA/89/003.

6. General Nutrition Consultancy

6.1 Field trip to shrimp feed mill and shrimp rearing areas in SW Thailand was conducted with Prasert Sitasit, GPC for project. The shrimp rearors cooperative feed mill near Surat Thani was examined. The capacity is about 4 tons per day of dried 2 mm pellets made by reconstitution of dry meals, premixes, oils and water, then extrusion through a meat grinder with 2 mm die, then tunnel drying before bagging. Capacity is sufficient for the local farmers involved but limited for much expansion.

6.2 The Fisheries Research and Development station at Sichon was visited. This large shrimp and marine fish larval rearing station has a fairly good marine water supply and many tanks which could be used for shrimp and marine fish diet trials during the off season when demand is light for shrimp post larvae production. The indoor tanks could be used for closely monitored pilot plant diet trials where response of animals would be dependent upon nutrients in the diets used without supplementation from pond water environment.

6.3 Recruitment procedures for temporary employees (NTE 3 months) was reviewed with FAO administrative personnel and THA/89/003 personnel.

6.4 Purchasing procedures for Field Purchase Orders and for UNDP Requisition for purchase was reviewed with THA/89/003 personnel. Imre Csavas, RAPA Regional Aquaculture Officer will approve by signature those FPO's and UNDPR's issued. FPO's are issued for outside country purchases and UNDPR's issued for local purchase of equipment and supplies.

6.5 Assistance in administrative clearance for fellowship training abroad was offered for the Ph.D. training programme at Mississippi State University for Nanthiya Unprasert. After several delays she was able to obtain all clearances and travel space for a last minute departure on 20 August to arrive at MSU 21 August at noon on last day of registration before classes started on 22 August. Contacts with Mr. McNaughton of IDRC were very productive and his rapid response with official support letters to DTEC enabled clearance and passport release in time. Professor Robert Wilson at MSU was also very helpful and understanding about the late arrival.

6.6 Contacts with Professors Shell and Grover at Auburn University resulted in acceptance of Munsiri as a M.Sc. program graduate student and Ms. Sonkphan Lumlertdacha as a special student pending completion of her GRE examination at Auburn before she will be accepted there for graduate study. Mr. J. M. Meyour, the Senior Fellowships Officer, FAO Rome was also very helpful and prompt in response to requests for early release of official award papers to expedite its clearances necessary by various offices in Thailand before passport and visa are obtained.

6.7 A list of catalogues, references and special supplies for the GLC and HPLC were assembled and filed in the office cabinets for Dr. Hardy's use on arrival.

7. RECOMMENDATIONS

7.1 Complete the sample preparation room next to the GLC - HPLC clean room. Install work bench and electrical outlets below window.

7.2 Rewire clean room with grounded labelled outlets above table inner wall.

7.3 Install water purification system for HPLC sample preparations.

7.4 Recruit and train research technician to operate GLC and HPLC equipment as sole duties under supervision of Dr. Wimol.

7.5 Have Mr. Pairat redesign coated vitamin C experiments with diets manufactured at monthly intervals and monitored for C levels during feeding trials.

7.6 For all diet trials with small fishes/shrimp have personnel feed early (0700) and late (1600 – 1700) daily to cover animal not human needs.

7.7 Purchase 6–10 Carousel type automatic feeders for diet trials in wet laboratory.

7.8 Have Dr. Wimol redesign and repeat protein requirement study with Pangasius catfish extending protein levels to 50 % of diet treatments (25, 30, 35, 40, 45, 50 % CP).

7.9 Purchase or requisition for purchase all remaining equipment immediately to allow time for delivery, installation and operation before end of project period.

7.10 Design Layout for extruder expanded pellet mill for line operation complementary to existing compressed pellet line.

7.11 Plumb boiler (to be ordered) into existing CPM mills.

7.12 Purchase another die and put second CPM mill into operating capability.

7.13 Clean service and connect existing standly generator to feed quality control and nutrition research laboratory for emergency use during long overnight type assays.

7.14 Adopt a major project proposal proforma system to plan and implement research effort according to documented priorities. See examples enclosed in THA/87/004 report of July 1989.

7.15 Develop cooperative use of facilities at Sichon research station for feeding trial testing on pilot scale diets formulated for marine shrimp and fish.

7.16 Request return of nutrition consultant for 1.5 man months during August - September 1991.

Personnel Contacted

Annex A

NATIONAL INLAND FISHERIES INSTITUTE - RESEARCH PROPOSAL

Amino acids and protein requirement of walking catfish (Clarias batrachus)

Introduction

Amino acids profile in fish feeds influence fish growth since fish do not have a true protein requirement, but need a well-balanced mixture of indispensable and dispensable amino acids (Wilson and Halver 1986). Fish, like other monogastric animals, require the same 10 indispensable amino acids : arginine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, and valine. Qualitative and quantitative requirement for these indispensable amino acids has been studied intensively in only four species ; chinook salmon (Halver 1972), common carp and Japanese eel (Nose and Arai, cited by Cowey and Sergent, 1979), and channel catfish (Wilson, et al 1977).

Walking catfish (Clarias batrachus) are one of the most raised fish in South-east Asia because of their fast-growing and ability to tolerant low oxygen conditions. The fish are scavengers and depend on high protein in their diet to grow. Protein requirements range from 25–35 % and DE/P range from 5.8-12 kcal/g (Amnuey and Weing 1982 ; Kuanruthai 1984 ; Banyat 1989). The inconsistency of protein requirements are due to the different protein sources. Therefore if the amino acids requirement were known, it could serve as basic information for formulation of Clarias feed regardless of the protein source.

Justification

Protein is the most expensive part in the fish diets. Protein is required for fish growth because it provides the mixture of amino acids necessary for protein synthesis. At present, Clarias diets are formulated to contain high protein levels (at least 30 %) to satisfy growth of the fish. However, the actual amino acids requirement by Clarias are not known. Therefore, the protein content recommended maybe too low to promote the full potential growth or maybe too high and cost more than it should. Qualitative and Quantitative requirements for amino acids by Clarias are needed to be the foundation for solving the problems associated with fish growth and feed cost. Amino acids requirements will be more universal than protein requirement by fish and will be useful for farmers scientists or feed formulators from different regions to establish standard feeds for Clarias.

Objectives

The objectives of this proposal are as following :

1. Determine the amino acids profiles in Clarias flesh and eggs.
2. Determine the amino acids requirement by Clarias using amino acids profiles in flesh and eggs as reference amino acids levels.
3. Protein digestibility of practical ingredient ; fish meal, soybean meal, rice bran and broken rice for Clarias.
4. Formulate Clarias production feed with protein level to satisfy all 10 indispensible amino acids and test in large scale ponds.

Procedure

Objective 1

Walking catfish and matured female walking catfish fed traditionally formulated commercial feed will be collected from commercial farms in central plain of Thailand and wild walking catfish and matured female walking catfish from natural waters will be purchased from fisherman. As soon as possible after the fish have reached the laboratory, egg from the matured female walking catfish will be removed. Carcass and egg of the walking catfish from each group will be analyzed for all indispensable amino acids. Amino acids will be determined in the National Inland Fisheries Institute by method of Chang et al. (1981) using high performance liquid chromatography.

Objective 2

Profiles of the indispensable amino acids in fish eggs and carcass will be used as references for amino acids requirements by fish. To determine each indispensable amino acid requirement, five basal diets will be formulated for each amino acid study. The diets compose of Casein (Vitamin-free), Gelatin, White dextrin, Cellulose, Corn oil, Fish oil, Vitamin mix, Mineral mix, CMC and Amino acid mixture. The diets will be isonitrogenous and isocaloric which contain 24 % crude protein and 275 Kcal DE/100 g diet (Wilson 1982). The basal diet in each study will be supplemented with graded levels of the amino acid to provide the appropriate test range under study.

The experimental diets will be fed to triplicated groups of fingerling walking catfish (2–5 cm in size) held in 80 liter glass aquaria for 10 weeks. Each group will be fed 3 % of body weight three times each day. The fish will be sampled biweekly, and the feed allowance will be adjusted accordingly. At the end of the experiment, growth curves of each amino acids study will be plotted. The break point where the growth rate leveled off will be considered as level required by the fish.

Objective 3

To formulate practical walking catfish feed to satisfy indispensable amino acid requirement using fish meal, soybean meal, rice bran and broken rice as main ingredients. Protein digestibility in these ingredients need to be determined. Each ingredient evaluated will be combined as a mixture of test diet which composed of reference diet, test ingredient, and chromic oxide. The protein digestibility of test diet will be compared to the protein digestibility reference diet and allow to calculate the protein digestibility of the ingredient. The procedure for protein digestibility determination will follow Brown et al. (1985).

Objective 4

When each of 10 indispensable amino acids required by walking catfish and protein digestibility of raw ingredients are known, practical diets will be formulated to have similar amino acids patterns as requirements. Formulated feed will compose of fish meal, soybean meal, rice bran and broken rice, and supplemented with crystalline amino acids if necessary. The requirement for amino acids will determine protein level in the feeds. Formulated feeds will be then evaluated under practical conditions with comparison with commercial floating pellet for walking catfish, traditional trash fish + rice bran diet, and NIFI walking catfish feed. Sixteen earthen ponds, 400 m² in area, each will be stocked with 10,000 fingerlings (5 cm in size). Group of four ponds each will be randomized for each of the diet evaluated. The fish will be fed at 3 % body weight twice daily at 6:00 AM and 6:00 PM for three months. Fish from each ponds will be sampled biweekly and adjusted for feed allowances. At the end of the experiment, the ponds will be drained and fish will be harvested. Twenty five fish from each pond will be collected for further determination of composition of gain.

Literature Review

Amino acid compositions influence the dietary protein requirement for fish (Wilson and Halver 1986). Quantitatively, amino acids requirements in fish have been established in only four species; chinook salmon, channel catfish, Japanese eel and common carp (NRC 1983). All fish seem to require 10 indispensable amino acids. Even one amino acid present in the protein at a level below that require by the fish prohibited growth (Halver 1987) in fish feeds.

The first successful amino acid test diet for fish was developed by Halver et al. (1957). They compared crystalline-L-amino acid supplemented diet referenced to the amino acid patterns of whole chicken egg protein, chinook salmon egg protein, and chinook yolk sac fry protein. The results indicated that the test diet based on whole chicken egg protein gave the best growth and feed efficiency for chinook salmon for a 12-week period. Ketola (1982) suggested that although the amino acid contents of fish eggs appear to differ from the reported dietary requirements of the fish, the composition of the egg provided an effective guide for successful amino acid supplementation of proteins in fish feeds for Atlantic salmon and rainbow trout.

Test diet of Halver et al. (1957) was unsuccessful in common carp for determination of amino acids requirement since a marked reduction in growth rate was observed corresponding to the relative amount of free amino acids in the test diets (Aoe et al 1970).

It has been suggested that for warm water fish such as channel catfish and common carp, the amino acids test diets must be neutralized with base for utilization by the fish (Dupree and Halver 1970; Nose et al. 1974). Wilson and Robinson (1982) showed that a significant improvement in crystalline amino acids used by channel catfish was observed by adjusting the pH of the test diets, and a diet of pH 7 produced a growth rate similar to the whole egg protein control diet.

Facilities and Equipment

Facility are available at the National Inland Fisheries Institute for preparing the sample for amino acids analysis and for preparing feeds. A high performance liquid chromatograph for amino acids analysis is also available. Sixty-80L-glass aquaria with two compressors for air supply are required. Sixteen-400 m² -earthen ponds will be available at Supanburi Fisheries Station. Walking catfish fingerlings need to be bought from hatchery. Purified ingredients and crystalline amino acids need to be ordered from the United States. Practical feed stuffs are locally available. Two technicians are required. One for HPLC operation and another for feed preparation and feeding.

Research Time Table

1. HPLC determination of amino acids profiles in walking catfish carcass and eggs.

2. Qualitative and Quantitative 10 indispensable amino acids requirement of walking catfish.

3. Protein digestibility study and practical walking catfish feed evaluation.

References

Amnuey C. and C. Weing. 1982. Protein requirement by walking catfish fry. Thai Fish. Gazzette 35:251–260.

Aoe, H., I. Masuda, I. Abe, T. Saito, T. Toyoda, and S. Kitamura 1970. Nutrition of Protein in young carp. I. Nutritive value of free amino acids. Bull. Jap. Soc. Sci. Fish. 36:407–413.

Banyat, S. 1989. Growth Respons of Walking Catfish Fingerling Fed Isocaloric Diets with Variable Protein Levels. Master Thesis. Kasetsart University.

Brown, P. B., R. J. Strange, and K. R. Robbins. 1985. Protein digestibility coefficients for yearling channel catfish fed high protein feedstuffs. Prog. Fish-Cult. 47:94–97.

Chang, J-Y, R. Knecht, and D. G. Brawn. 1981. Amino acid analysis at the picomole level: Application to the C-terminal sequence analysis of polypeptide. Biochem J. 199:547.

Cowey, C. B. and J. R. Sargent. 1979. Nutrition P. 1–69 in Fish Physiology, Bioenergetics and Growth, Vol. VIII, W. S. Hoar, D. J. Randall, and J. R. Brett, (eds.). Academic Press. New York.

Dupree H. K. and J. E. Halver 1970. Amino acids essential for the growth of channel catfish. Trans. Am. Fish. Soc. 99:90–92.

Halver, J. E. (Ed). 1972. Fish Nutrition. Academic Press, New York.

Halver, J. E. (Ed). 1987. Fish Nutrition. Second Edition. Academic Press.

Halver, J. E., D. C. Delong, and E. T. Mertz 1957. Nutrition of salmonoid fishes. V. Classification of essential amino acids for chinook salmon. J. Nutr. 63:95–105.

Ketola, H. G. 1982. Amino acid nutrition of fishes: requirements and supplementation of diets. Comp. Biochem. Physiol. 73B:17–24.

Kuanruthai, T. 1984. Effect of Isonitrogenous Feed with Different Energy Levels on Growth and Surviving of Walking Catfish Fry. Master Thesis. Kasetsart University.

Nose T., S. Arai, D-L Lee, and Y. Hashimoto 1974. A note on amino acids essential for growth of young carp. Bull. Jap. Soc. Scient. Fish 40:903– 908.

NRC 1983. Nutrient Requirements of Warmwater Fishes and Shellfishes. National Academy Press. Washington D. C.

Wilson, R. P., D. E. Harding, and D. L. Garling, Jr. 1977. Effect of dietary pH on amino acid utilization and the lysine requirement of fingerling channel catfish. J. Nutrition 107:166–170.

Wilson, R. P., and J. E. Halver (1986). Protein and amino acid requirements of fishes. Ann. Rev. Nutr. 6:225–244.

Wilson, R. P. and E. H. Robinson 1982. Protein and Amino Acid Nutrition for Channel Catfish. Mississippi Agricultural and Forestry Experiment Station.

Budget (5 years duration proposed)

ALLOCATIONS REQUESTED

Annex B

Principles of Fish Nutrition

Annex C

LIST OF PARTICIPANTS IN THE IN-SERVICE TRAINING DURING 7–10 AUGUST 1990

Annex E

Protocol for C₁ and Bound C Assays

1. Weigh 1 g wet tissue into test tube.

2. Add 9 mL Cold 5 % Trichloracetic acid solution.

3. Homogenize in Polytron 1 min advancing to high speed.

4. Transfer to clinical centrifuge.

5. Spin 5–10 min hi speed angle head.

6. Take 2 mL of supernatant for A and B.

7. For A

   1. Add 1 mL color reagent;
   2. Incubate at 37° C one hour;
   3. Chill in ice bath;
   4. Add 2 mL 75% H₂SO₄ slowly;
   5. Mix and let stand 20–30 minutes;
   6. Read @ 515 mm in spectrophotometer.

   Compare with standard curve of 5 – 30 (Mg) C₁/mL.

   For B

   1. Add 1 mL color reagent;
   2. Hold in water bath at 90–100° C for 20 minutes;
   3. Chill in ice bath;
   4. Add 2 mL 75% H₂SO₄ slowly;
   5. Let stand 20–30 minutes;
   6. Read @ 515 mm in spectro photometer.

   Reads Total C in sample.
   Bound C = Total C - C₁

Color Reagent

1 mL 0.6% CuSO₄
1 mL 5% Thiourea
20 mL 2.2%, 2, 4-Dinitrophenylhydrazine solution in 25% H₂SO₄
Mix reagents daily - Color mix will hold in refrigerator for 1 day.

Note : Make up all C₁ standard solutions in 5 % TCA.

Annex F

Tucker - Halver Modification of C Analysis

Reagents :

75 % H₂SO₄
9N (25 %) H₂SO₄
DNPH : 2.2 % in 9NH₂SO₄
CuSO₄ : 0.5 g CuSO₄.5H₂O in 100 mL HOH
Thiourea : 1.0 g Thiourea in 100 mL . HOH
DTC reagent : Prone to selfoxidation
Mix 10 DNPH : 1 Thiourea : 1 CuSO₄ Solutions daily
TCA : 6 % Trichloracetic acid in HOH

Stock standards - good for one week in refrigerator
C₁ (MW 176) : 20 mg made to 100 mL with 5 % TCA
C₂ (MW 368) : 21 mg made to 50 mL with HOH
Dilute to concentrations desired (20 ug C₁ equivalent/mL)
Develop standard curve 5 – 25 ug/mL C₁

Procedure :

1. Extract 1–2 g Liver or 0.5 g feed + 9 parts 6 % TCA with polytron homegenizer for 1 minute.
2. Spin down in clinical centrifuge at high speed for 3–5 minutes. Save supernatant.
3. Take 1 mL. aliquote for Sample A Take 1 mL. aliquote for Sample B
4. Add 0.3 mL Color Reagent to A and B
5. Incubate Sample A 2–3 hrs @ 37° C
6. Treat Sample B at 95–100° C for exactly 20 minutes
7. Chill Sample B
8. Add 1.7 mL 75 % H₂SO₄
9. Chill Sample B
10. Let Stand 20–30 minutes @ room temperature
11. Read in Spectrophotometer at 515 nm = Total C
12. Add 1.7 mL 75 % H₂SO₄ to Sample A
13. Chill Sample A
14. Let stand 20–30 minutes @ room temperature
15. Read in Spectrophotometer at 515 nm = C₁

    Total C - C₁ = bound C

Annex G

Protocol for C₁, C₂, C₃ Assays

1. Weigh 0.50–1.00 g liver tissue or feed to plastic test tube.

2. Calculate weight and dilute W/ 9 parts water.

3. Homogenize one minute in Polytron.

4. Transfer homogenate to 100 mL glass beaker W/ cover.

5. Micro wave one minute. Cool in icebath.

6. Transfer beaker contents to centrifuge tube.

7. Centrifuge 3 min hi speed in clinical centrifuge.

8. Decant and save supernatant in icebath.

9. Add calculated 5 mL water to pellet in tube.

10. Homogenize one minute in Polytron.

11. Transfer to same 100 mL beaker W/ cover glass.

12. Microwave homogenate one minute ; Cool.

13. Transfer to centrifuge tube.

14. Spin 3 min hi speed in clinical centrifuge.

15. Decant - off supernatant. Combine with first.

16. Mix supernatants on Vortex mixer; Place in icebath.

17. Remove 1–2 mL; Pressure filter thru 0.45 um disk.

18. Collect 1–1.5 mL filtrate.

19. Inject 25–50 uL into HPLC.

20. Read C₁, C₂, C_x, C₃ etc.

21. Calculate Quantitative yield from aliquot used.

Annex H
Equipment Needed for Analysis

1. Clinical centrifuge with angle head 18 mm tubes.

2. Spectrophotometer, double beam, UV and visible light.

3. Protein hydrolysis tubes - 12.

4. Adjustable auto pipettes 0.1 – 1.0 mL size.

5. Tips for above, 200

6. Purified water supply for HPLC - - ordered.

7. Cuvettes for spectrophotometer 2 mL quartz - 4

8. Heating block, 12 space, 18 mm.

9. Test tubes 18 × 100 mm. - 100.

10. Separatory funnels 500 mL - 10.

11. Separatory funnels 250 mL - 10.

12. Water aspirator suction unit 3.

13. Voltage Regulator for HPLC.

Annex J
LIPIDS IN CRUSTACEAN NUTRITION

BIBLIOGRAPHY COMPILED

BY

Dr. LOUIS R. D'ABRAMO,
Dept. Wildlife and Fisheries, Mississippi State University,
P.O. Drawer LW, Mississippi State, MS 39762, USA.

and

Dr. John D. Castell,
Invertebrate Nutrition Group Leader, Dept. of Fisheries & Oceans, Biol. Sci. Br.,
Benthic Fish & Aquacult. Div., P.O. Box 550, Halifax, Nova Scotia, B3J 2S7, Canada.

Ackman, R.G.; Eaton, C.A.; Sipos, J.C.; Hooper, S.N.; Castell, J.D. 1970. Lipids and Fatty Acids of Two Species of North Allantic Krill (Meganyctiphanes norvegica and Thysanoessa inermis) and Their Role in the Aquatic Food Web. J. Fish. Res. Bd. Can. 27(3): 513–533.

Addison, R.F.; Ackman, R.G.; Hooper, S.N.; Hingley, J.; Regier, L.W. 1970. Lipid Content and Composition of Queen Crab (Chionoectes opilio) Tissue and Products. Fish. Res. Bd. Can. Tech. Rep. #198.

Ando, T.; Kanazawa, A.; Teshima. S.; Patrois, J.; Ceccaldi, H.J. 1977. Variation in the Lipids of Tissues During the Molt Cycle of Prawn. Bull. Jpn. Soc. Sci. Fish. 43(12): 1445–1449.

Bianchini, M. 1984. Effect of cholesterol in artifical diels for Mediterranean prawns Quad. Ist. ldrobiol. Acquacoll. 4: 16–25.

Bianchini, M.L. 1984. Effects of Cholesterol in Artificial Diets for Mediterranean Prawns Proc. 1st Intern. Cont. Cult. Penaeid Prawns/Shrimp 1985. P. 183.

Bottino, N.R. 1974. The Fatty Acids of Antartic Phytoplankion and Euphauslids. Fatty Acid Exchange among Trophic Levels of the Ross Sea. Mar. Biol. 27: 197–204.

Bottino, N.R.; Gennity, J.; Lilly, M.L.; Simmons, E.; Finne. G. 1980. Seasonal and Nutritional Effects on the Fatty Acids of Three Species of Shrimp, Penaeus setiferus, P. aztecus and P. duorarum. Aquaculture

Bottino, N.R.; Lilly, M.L.; Finne, G. 1979. Fatty Acid Stability of Gulf of Mexico Brown Shrimp (Penaeus aztecus) Held on Ice and in Frozen Storage. J. Food Sci. 44: 1778– 1779.

Bourdier, G.G.; Amblard, C.A. 1989. Lipids in Acanthodiaptomus denticornis during Starvation and Fed on Three Differant Algae J. Plankton Res. 11(6): 1201–1212.

Briggs, M.R.P.; Jauncey, K.; Brown, J.H. 1988. The Cholesterol and Lecithin Requirements of Juvenile Prawn (Macrobrachium rosenbergii) Fed Semi-Purified Diets Aquaculture 70(½): 121– 129.

Brockerhoff, H.; Stewart, J.E.; Tacreiter, W. 1967. Digestion of Triglycerides by Lobster. Can. J. Biochem. 45: 421–422.

Cahu, C.; Fauvel, C. 1986. Effect of Food Fatty Acid Composition of Penaeus vannamei Broodstock on Egg Quality ICES-CM-1986/F: 28 F:28

Cahu, C.: Quazuguel. P. 1989. Lipid Metabolism of Penaeus vannamei Broodstock: Influence of Dietary Lipids. Aquaculture Europe '89, International Aquaculture Conference. Bordeaux (France). R. Billard and N. De Pauw European Aquaculture Society Special Publication No. 10 p. 45–50.

Casteil, J.D. 1967. Preliminary Data on Lipid Composition of Oregon Shrimp. Unpublished Data? 1967

Castell, J.D. 1981. Fatty Acid Metabolism in Crustaceans. In: Proc. 2nd International Conference on Aquaculture Nutrilion: Blochemical and Biophysiological Approaches to Shellfish Nutrition. G.D. Pruder, C. Langdon and D.E. Conklin World Maricutt Soc. Spec. Pub. #2 pp. 124–145.

Castell, J. D.; Boghen, A.D. 1979. Fatty Acid Melabolism in Juvenile Lobslers (Homarus americanus) Fed a Diet Low in Methionine and Histidine. Proc. World Maricult. Soc. 10: 720–727.

Castell, J.D.; Covey. J.F. 1976. Dielary Lipid Requirements of Adult Lobsters, Homarus americanus (M.E.). J. Nutr. 106(8): 1159–1165.

Castell, J.D.; Mason, E.G.; Covey, J.F. 1975. Cholesterol Requirements of Juvenile Lobster (Homarus americanus). J. Fish. Res. Bd. Can. 32(8): 1431–1435.

Cataculan, M.R.; Kanazawa, A. 1985. Effect of Dietary Fatty Acids on the Fatty Acid Composition of Penaeus monodon Juveniles. Proc. 1st Intern. Conf. Cutt. Penaeld Prawns/Shrimp 1985. p. 181.

Chanmugam, P.; Donovan, J.; Wheelwe, C.J.; Hwang, D.H. 1983. Differences in the Lipid Composition of Freshwater Prawn (Macrobrachium rosenbergh) and Marine Shrimp J. Food Sci. 48: 1440–1443.

Chapelle, S. 1986. Aspects of Phospholipid Metabolism in Cruslaceans as Related to Changes in Environmental Temperatures and Salinlties. Comp. Biochem. Physiol. 84B: 423–439.

Chapelle. S.; Meister, R.; Brichon, G.; Zwingelstein, G. 1977. Influence of Temperature on the Phospholipid Metabolism of Various Tissues from the Crab Carcinus meanas. Comp. Biochem. Physiol. 58B: 413–417.

Chapelle, S. 1978. The Influence of Acclimation Temperature on the Fatty Acid Composition of an Aquatic Cruslacean (Carcinus maenus). J. Exp. Zool. 204: 337– 346.

Clarke, A. 1979. Lipid Content and Composition of the Pink Shrimp, Pandalus montagui (Leach) (Crustacea : Decapoda). J. Exp. Mar. Biol. Ecol. 38(1): 1–17.

Clarke, A. 1982. Lipid synthesis and Reproduction in the Polar Shrimp Chorismus antarcticus. Mar. Ecol. (Prog. Ser.) 9(1): 81–90.

Clarke. A. 1984. The Lipid Content and Composition of Some Antarctic Macrozooplankton. Br. Antarct. Surv. Bull. 63: 57–70.

Clark, A.; Wickins, J.F. 1980. Lipid Content and Composition of Cultured Penaeus merguiensis fed with Animal Food Aquaculture 22(1): 17–27.

Claus, C.; Benijts, F.; Yandeputle, G.; Gardner, W. 1979. The Biochemical Composition of the Larvae of Two Strains of Artemia salina (L.) Reared on Two Different Algae Foods. J. Exp. Mar. Biol. Ecol. 36: 171–183.

Collatz, K-G. 1969. The Fatty Acid Spectrum In the Crayfish, Orconectes limosus, and its Dependence on Diel. J. Comp. Physiol. 65: 291–298.

Collatz, K-G. 1969. The Lipid Spectrum of the Crayfish, Orconectes limosus, and its seasonal changes. J. Compar. Physiol. 65: 274–290.

Colvin, P.M. 1976. The Effect of Selected Seed Oils on the Fatty Acid Composition and Growth of Penaeus indicus. Aquaculture 8(1): 81–89.

Conklin, D.E.; D'Abramo, L.R.; Bordner, C.E.; Baum, N.A. 1980. A Successful Purified Diet for the Culture of Juvenile Lobsters; The Effects of Lecithin. Aquaculture 21: 243–249.

Croz, L; Wong, L.; Justine, G.; Gupia, M. 1988. Prosiaglandins and Related Compounds from the Polychaele Worm Americonuphis reesel Fauchald (Onuphidae) as Possible Inducers of Gonadal Maluration in Penaeid Shrimp. Rev. Biol. Trop. 36(2A): 331–332.

Cuzon, G.; Cahu, C.; Aldrin, J.F.; Messager, J. L.; Stephan, G.; Mevel, M., 1980. Starvation Effect on Metabolism of Penaeus laponicus. Proc. World Maricutt. Soc. 11: 410–423.

D'Abramo, L.R. 1979. Dietary Fatty Acid and Temperature Effects on the Productivity of the Cladoceran, Moina macrocopa. Biol. Bull. 157(2): 234–348.

D'Abramo, L.R. Baurn, N.A.; Bordner, C.E.: Conklin, D.E.; Chang, E.S. 1985. Diet-Dependent Cholesterol Transport in American Lobster. J. Exp. Mar. Biol. Ecol. 87: 83–96.

D'Abramo. L.R.; Bordner, C.E.; Conklin, D.E. 1982. Relationship Between Dietary Phosphalidyicholine and Serum Cholesterol in the Lobster Homarus sp. Marine Biol. 67: 231–235.

D'Abramo, L.R.; Bordner, C.E.; Conklin, D.E.; Baum. N.A. 1981. Essentiality of Dietary Phosphalidyicholine for the Survival of Juvenile Lobsters. J. Nutr. 111: 425–431.

D'Abramo. L.R.; Bordner, C.E.; Conklin, D.E.; Baum, N.A. 1984. Sterol Requirement of Juvenile Lobsters, Homarus sp. Aquaculture 42(1): 13–25.

D'Abramo. L.R.; Bordner, C.E.; Dagelt. G.R.; Conklin, D.E.; Baum, N.A. 1980. Relationship Among Dietary Lipids, Tissue Lipids, and Growth in Juvenile Lobsters. Proc. World Maricult. Soc. 11: 335–345.

D'Abramo, L.R.; Wright, J.S.; Wright, K.H.; Bordner, C.E.; Conklin, D.E. 1985. Sterol Requirement of Cultured Juvenile Crayfish Moina macrocopa. Aquaculture 49: 245– 255.

Dall, W. 1981. Lipid Absorption and Utilization in the Norwegian Lobster, Nephrops norvegicus L. J. exp. mar. Biol. Ecol. 50: 33–45.

Davis, D.A.: Robinson, E.H. 1986. Estimation of the Dietary Lipid Requirement Level of the White Crayfish Procambarus acutus. J. World Aquacult. Soc. 17: 37–43.

Deshimaru, O.; Kurold, K.; Yone, Y. 1979. The Composition and Level of Dietary Lipid Appropriate for Growth of Prawn. Bull. Jpn. Soc. Sc. Fish. 45(5): 591–595. Deshimuru, O.; Kuroki, K. 1974. Studies on a Purified Diet for Prawn - II. Optimum Contents of Cholesterol and Glucosamine in the Diet. Bull. Jpn. Soc. Sci. Fish. 40(4): 421–424.

Farkas, T. 1970. Fatty Acid Composition of Lipids Obtained from Eudiaptomus gracilis G. O. Sars (Copepoda) and Daphnia cucullata G.O. Sars (Cladocera). Acta. biol. Acad. Sci. Hung. 21(2): 225–233.

Farkas, T.: Herodek, S. 1964. The Effect of Environmental Temperature on the Fatty Acid Composition of Crustacean Plankton. J. Lipid. Res. 5: 369–373.

Fenucci, J.L.; Lawrence, A.L.; Zein-Elden, Z.P. 1981. The Effects of Fatty Acid and Shrimp Meal Composition of Prepared Diets on Growth of Juvenile Shrimp. Penaeus stylirostris. J. World Maricult. Soc. 12: 315–324.

Galois, R., 1987 1987. Neutral Lipids in Decapod Crustaceans: Metabolism and Requirements. (1976–1986): Ten Years of Research in Aquaculture. Part 2: The Crustaceans. Laubler, L. Oceanis 13(2): 197–215.

Galois, R.G. 1984. Variations in Tissue Lipid Composition During Vilellogenesis in the Prawn Penaeus Indicus Milne Edwards. J. Exp. Mar. Biol. Ecol. 84(2): 155–166.

Gardner, D.; Riley, J.P. 1972. Seasonal Variations in the Component Fally Acid Distributions of the Lipids of Balanus balanoides. J. Mar. Blol. Ass. U.K. 52: 839–845.

Gonzalez-Baro, M.D.R.: Pollero, R.J. 1988. Lipid Characterization and Distribution among Tissues of the Freshwater Crusiacean Macrobrachium boreliff During an Annual Cycle. Comp. Biochem. Physiol. 91B(4): 711–716.

Gopakumar, K.; Nair, M.R. 1975. Lipid Composition of Five Species of Indian Prawns. J. Sci. Fd. Agric. 26: 319–325.

Guary, J.C.B.; Kanazawa, A. 1973. Distribution and Fale of Exogehous Cholesterol during the Molting Cycle of the Prawn, Penaeus japonicus Bale. Comp. Biochem. Physiol. 46A(1): 5–10.

Guary, J.C.; Kayama, M.; Murakami, Y.; Ceccaldi, H. 1976. The Effects of a Fat-Free Diet and Compounded Diets Supplemented with Various Oils on Moli, Growth and Fatty Acid Composition of prawn Penaeus japonicus Bale Aquaculture 7; 245–254.

Guary, J-C.; Kayama. M.; Murakami, Y. 1974. Lipid Class Distribution and Fally Acid Composition of Prawn, Penaeus japonicus Bale. Bull. Jpn. Soc. Sci. Fish. 40(10): 1027–1032.

Guary, J-C.: Kayama, M.; Murakami, Y. 1975. Seasonal Variation of the Fally Acid Composition of Penaeus japonicus (Crusiacea : Decapoda). Mar. Biol. 29(4): 335–342.

Hayashi, K. 1976. The Lipids of Marine Animals from Various Habital Depths. VI. On the Characteristics of the Fally Acid Composition of Neutral Lipids from Decapods. Bult. Fac. Fish., Hokkaido Univ 27(1): 21–29.

Herodek, S. 1970. Desaturation of Palmitic Acid-1-C14 and Slearic Acid-1-C14 in Gammarus (Rivulogammarus) Roeselii gervais (Crustaces, Amphipods). Annal.

Hilton, J.W.: Harrison, K.E.; Slinger, S.J. 1984. A Semi-purified Test Diet for Macrobrachium rosenbergii and the Lack of Need for Supplemental Lecithin. Aquaculture 37: 209-

Hinchclille, P.R.: Riley, J.P. 1972. The Effect of Diet on the Component Fatty Acid Composition of Artemia salina. J. Mar. Blol. Ass. U.K. 52: 203–211.

Idler, D.R.: Wiseman. D. 1971. Sterols of Crustaceans. Intern. J. Biochem. 2(7): 91– 98.

Jeckel, Walter H.; de Moreno, Julia E.A.: Moreno, V.J. 1989. Biochemical Composition, Lipid Class and Fatty Acids in the Ovary of the Shrimp Pleoticus muetteri Bale. Comp. Biochem. Physiol. 92B(2): 271–276.

Jeckel, Walter H.: de Moreno, Julia E.A.: Moreno, Victor J. 1989. Biochemical Composition, Lipid Classes and Fatty Acids in the Male Reproductive System of the Shrimp Pleoticus multeri Bale. Comp. Biochem. Physlol. 93B(4): 807–811.

Jones, D.A.; Kanazawa, A.; Ono, K. 1979. Studies on the Nutritional Requirements of the Larval Stages of Penaeus japonicus Using Microencapsulated Diets. Marine Biology 54: 261–267.

Joseph, J.D.: Williams, J.E. 1975. Shrimp Head Oil: A Potential Feed Additive for Mariculture. Proc. World Maricult. Soc. 6: 147–155.

Kal, Hikaru: Kanazawa, A. 1989, Optimum Contents of Cholesterol in the Purified Diel for Grass Prawn Penaus monodon Abstracts of the Second Asian Fisheries Forum OC 016 pp. 68.

Kanazawa, A. 1977. Role of Lipids in Crustaceans (Review)(in Japanese).(Can. Trans. Fish. Aq. Sci. 4695) Feed Oll Abstracis 1276(B4): 5–10.

Kanazawa. A. 1983. Penaeld Nutrition. In: Proceedings of the Second International Conference on Aquaculture Nutrition Biochemical and Physiological Approaches to Shelfish Nutrition. G.D. Pruder, D.E. Conklin and C. Langdon (Eds.) World Maricult. Soc. Special Publication No. 2 pp.

Kanazawa, A. 1987. Turnover of Dietary Cholesterol and Sitosterol in the Prawn. Bull. Jpn. Soc. Sci. Fish. 53: 601–606.

Kanazawa, A.; Chim, L.; Laubler, A. 1988. Tissue Uptake of Radioactive cholesterol in the Prawn Penaeus japonicus Bate During Induced Ovarian Maturation Aquat. Living Resour. 1(2): 85–91.

Kanazawa, A.: Guary, J-C.B.: Ceccaidi, H.C. 1976. Metabolism of (14C) Beta -Sitosterol Injected at Various Stages of the Molling Cycle in Prawn Penaeus Japonicus Bate. Comp. Biochem. Physiol. 54B: 205–208.

Kanazawa, A.: Tanaka, N.: Kashiwada, K. 1972. Nutritional Requirements of Prawn-IV. The Dietary Effects of Ecdysones. Bull. Jpn. Soc. Sci. Fish. 38(9): 1067–1071.

Kanazawa, A.: Tanaka, N.: Teshima, S.: Kashiwada, K. 1971, Nutritional Requirements of Prawn - III. Utilization of the Dietary Sterois. Bult. Jpn. Soc. Sci. Fish. 37: 1015–1019.

Kanazawa, A.: Tanaka, N.; Teshima, S.: Kashiwada, K. 1971 Nutritional Requirements of Prawn - II. Requirements fro Sterois. Bult. Jpn. Soc. Sci. Fish. 37: 211–215.

Kanazawa, A.: Teshima, S. 1971. In Vivo Conversion of Cholesterol to Steroid Hormones in the Spiny Lobster, Panulirus Japonicus. Bull. Jpn. Soc. Sci. Fish. 37: 891–896.

Kanazawa, A.: Teshima, S. 1977. Biosynthesis of Fatty Acids from Acetate in the Prawn, Penaeus japonicus. Mem. Fac. Fish., Kagoshima Univ. 26: 49–53.

Kanazawa, A.: Teshima, S.: Endo, M. 1979. Requirements of Prawn, Penaeus japonicus, for Essential Fatty Acids. Mem. Fac. Fish., Kagoshima Univ. 28: 27–3.

Kanazawa, A.: Teshima, S.: Endo, M.: Kayama, M. 1978. Effects of Elcosapenfenoic Acid on Growth and Fatty Acid Composition of the Prawn, Penaeus japonicus. Mem. Fac. Fish., Kagoshima Univ. 27(1): 35–40.

Kanazawa, A.: Teshima, S.: Ono, K. 1979. Relationship Between Essential Fatty Acid Requirements of Aquatic Animals and the Capacity for Bioconversion of Linolenic Acid to Highly Unsaturated Fatty Acids. Comp. Biochem. Physiol. 63B: 295–297.

Kanazawa, A.: Teshima, S.: Ono, K.: Chalayondeja, K. 1979. Biosynthesis of Fatty Acids from Acetate in the Prawns. Penaeus monodon and Penaeus merguiensis. Mem. Fac. Fish., Kagoshima Univ. 28: 21-

Kanazawa, A.: Teshima. S.: Sakamoto, M. 1985. Effects of Dietary Lipids, Fatty Acids and Phospholipids on Growth and Survival of Prawn (Penaeus japonicus) Larvae. Aquaculture 50: 39–49.

Kanazawa, A.: Teshima, S.: Sakamolo, Y. 1975. Utilization of Dietary Cholesterol during the Molting Cycle of Prawn. Bull. Jpn. Soc. of Sci. Fish. 41(11): 1185–1189.

Kanazawa, A.; Teshima, S.: Sakamoto, Y.: Guary, J.-C.B. 1976. The Variation of Lipids and Cholesterol Contents in the Tissues of Prawn, Penaeus japonicus, During the Molt Cycle. Bult. Jpn. Soc. Sci. Fish. 49(9): 1003–1007.

Kanazawa, A.; Teshima, S.: Tokiwa, S. 1977. Nutritional Requirements of Prawn-VII Effect of Dietary Lipids on Growth. Bull. Jpn. Soc. Sci. Fish. 43(7): 849–856.

Kanazawa, A; Teshima, S.: Tokiwa, S. 1979. Biosynthesis of Fatty Acids from Palmitic Acid in the Prawn, Penaeus japonicus. Mem. Fac. Fish., Kagoshima Univ. 28: 17– 20.

Kanazawa, A.: Teshima, S.: Tokiwa, S.: Ceccaldi, H.J. 1979. Effects of Dietary Linoleic and Linolenic acids on Growth of Prawn. Oceanologica Acid 2(1): 43–47.

Kanazawa, A.: Teshima, S.: Tokiwa, S.: Endo, M.: Razek, F.A.A. 1979. Effects of Short-Necked Clam Phospholipids on the Growth of Prawn. Bult. Jpn. Soc. Sc. Fish. 45(8): 961–965.

Kanazawa, A.: Teshima, S.: Tokiwa, S.; Kayama, M.: Hirata, M. 1979. Essential Fatty Acids in the Diet of Prawn-II Effect of Docosahexaenoic Acid on Growth. Bult. Jpn. Soc. Sc. Fish. 45(9): 1151–1153.

Kanazawa, A.; Teshima, S.: Tokiwa, S.: Kayama, M.; Hirala, M. 1979. Essential Fatty Acids in the Diet of Prawn-II Effect of Docosahexaenoic Acid on Growth. Bull. Jpn. Soc. Sc. Fish. 45(9): 1151–1153. Kanazawa, A.; Tokiwa, S.; Kayama, M.; Hirata, M. 1977. Essential Fatty Acids in the Diet of Prawn-I Effects of Linolelc and Linolenic Acids on Growth. Bull. Jpn. Soc. Sc. Fish. 43(9): 1111–1114.

Kaler, S.B.: Spazianl, E. 1971. Incorporation of (14C)-Cholesterol into Crab Cholesterol Pools and Eodysones as a Function of the Molting Cycle. Am. Zool. 11:672-

Kattner, G.: Krause, M. 1989 Seasonal Variation of Lipids, Wax Esters, Falty Acids and Alcohols in Calanold Copepods from the North Sea. Mar. Chem. 26(3): 261–276.

Kawada, K.; Seino, H.; Walanabe, S.; Abe, Y. 1973. Study on the Liver Oil of Marine Crustaces Il. Composition of Fatty Acids and Components of Unsaponifiable Matters in the Liver Oil of Chionaecetes opilis. Lithodes turritus and Paralithodes camischatica. Yukagaku (Oil Chemistry) 22(1): 654–659.

Kayama, M.; Hirala, M. 1984. Fatty Acid Metabolism in Blue Crab, Portunus trituberculatus. with Special Reference to de novo Synthesis and Conversion. Yukzyahu 33: 16–24.

Kayama. M.; Hirala, M.; Kanazawa, A.; Tokiwa, S.; Saito, M. 1980. Essential Fatty Acids in the Diet of Prawn - III Lipid Metabolism and Fatty Acid Composition. Bull. Jpn. Soc. Sci. Fish. 46(4): 483–488.

Kayama. M.; Tsuchiya. Y.; Mead, J.F. 1963. A Model Experiment of Aqualic Food Chain With Special Significance in Fatty Acid Conversion. Bult. Jpn. Soc. Sci. Fish. 29: 452–458.

Kean, J.C.; Castell, J.D.; Boghen, A.G.: D'Abramo, L.R.; Conklin, D.E. 1985. A Re-Evaluation of the Lecithiri and Cholesterol Requirements of Juvenile Lobster (Homarus americanus) using Crab Protein-Based Diets. Aquaculture 47: 143–149.

Krishnamoorthy, R.V.: Lakshmi, G.L.: Ventkatamiah, A. 1980. Synthesis in vitro of Cholesterol by Milochondria in the Shrimp Penaeus aziecus (Ives). J. Biol. Sci. 2: 121–127.

Krishnamuorthy, R.V.; Venkataramiah, A.; Lakshmi, G.J.; Biesol, P. 1982. Effects of Starvation and Algae-Feeding on the Tissue Cholesterol Levels in Commercial Shrimp Penaeus aziecus Ives. Proc Symp. Costal Aquacult. 1: 215–222.

Krzynowek, J.; Wiggin. K.; Dohahue. P. 1982. Cholesterol and Fatty Acid Content in Three Species of Crab Found in the Northwest Atlantic. J. Food Sci. 47: 1025–1026.

Kuo, H.C.; Lee, T.C.; Chichester, C.O.; Simpson, K.L. 1976. The Carolenolds in the Deep Sea Red Crab. Geryon quinquedens. Comp. Biochem. Physiol. 54B: 387–390.

Lal, B.; Singh, T.P. 1987. Impact of Pesticides on Lipid Metabolism in the Freshwater Crayfish Clarias batrachus. during the Vitellogenic Phase of its Annual Reproductive Cycle. Ecoloxicol. Environm. Safety 13: 12–23.

Lautier, J.; Lagarrigue, J-G. 1987. Lipid Metabolism of the Female Crab Paghygrapsus marmoratus During the Molting Cycle. Biochem. Syst. Ecol. 15(5): 611–620.

Lautier, J.; Lagarrihue, J-G. 1988. Lipid Metabolism of the Crab Pachygrapsus marmoratus During Vitellogenesis. Biochem. Syst. Ecol. 16(2): 203–212.

Lester, R.; Carey. M.C.; Little, J.M.; Dowd. S.R. 1975. Crustacean Intestinal Delergent Promotes Sterol Solubilization. Science 189: 1090–1100.

Levine, D.M.; Sulkin, S.D. 1984. Nutritional Significance of Long-Chain Polyunsaturated Fatty Acids to the Zoeal Development of the Brachyuran Crab. Eurypanopeus depressus (Smith). J. Exp. Mar. Ecol. 81: 211–223.

Martin, B.J. 1980. Growth and Fatty Acids of Palaemon serratus Fed with Compounded Diets Containing Different Proportions of Linoleic and Linolenic Acids. Aquaculture 19: 325–337.

Martin, B.J.; Ceccaldl, H.J. 1977. Influence of Temperature on the Fatty Acid Composition of Abdominal Muscle of Palaemon serr atus. Biochem. Syst. Ecol. 5: 151–154.

Middledltch, B.S.; Missler, S.R.; Hines, H.B. 1980. Metabolic Profiles of Penaeld Shrimp: Dietary Lipids and Ovarian Maturation. J. Chromatography 195: 359– 368.

Middleditch, B.S.; Missler, S.R.; Hines, H.B.; Chang, E.S.; McVey, J.P.; Brown, A.; Lawrence, A.L. 1980. Maturation of Penaeld Shrimp: Lipids in the Marine Food Web. Proc. World Maricult. Soc. 11: 463– 470.

Middleditch, B.S.; Missler, S.R.; Hines, H.B.; McVey, J.P.: Brown, A.; Ward, D.G.; Lawrence, A.L. 1980. Metabolic Profiles of Penaeld Shrimp; Dietary Lipids and Ovarian Maturation. J. Chromatog. 195: 359–368.

Middleditch, B.S.; Missler, S.R.; Ward, D.G.; McVey, J.B.; Brown, A.; Lawrence, A.L. 1979. Maturation of Penaeld Shrimp: Dietary Fatty Acids. Proc. World Maricult. Soc. 10: 472–476.

Milliamena, O.L.; Ouinttlo, E.T. 1985. Lipids and Essential Fatty Acids in the Nutrition of Penaeus monodon Larvae. Proc. 1st Interm. Cont. Cult. Penaeld Prawns/Shrimp. p. 181.

Millamena, O.M.; Pudadera, R.A.; Calacutan, M.R. 1985. Effects of Diet on Reproductive Performance of Abiated Penaeus Monodon Broodstock Proc. 1st Int. Conf. Cult. Penaeld Prawns pp. 178–179.

Morris, R.J. 1973. Relationships Between the Sex and Degree of Maturity of Marine Crustaceans and Their Lipid Composition. J. Mar. Biol. Ass. U.K. 53: 27–37.

Morris, R.J.; Ferguson, C.F.; Raymont, J.E.G. 1973. Preliminary Studies on the Lipid Metabolism of Neomysis Integer, Involving Labelled Feesing Experiments. J. Mar. Biol. Ass. U.K. 53: 657–664.

Morris, R.J.: Sargent, J.R. 1973. Studies on the Lipid Metabolism of Some Oceanic Crustaceans. Mar. Biol, 22: 77–83.

Nakagawa, H.: Ceccaldl, H.J. 1985. Circadian Variations of Haemolymph Lipoprotein of Palaemon serratus. Biochem. Syst. Ecology 13(3): 345–348.

O'Connor, J.D.: Gilbert, L.I. 1969. Alterations in Lipid Metabolism Associated with Premolt Activity in a Land Crab and Fresh-Water Crayfish. Comp. Biochem. Physiol. 29: 889–904.

Olazu-Abrill, M.: Martin, B.J.: Ceccaldi, H.J. 1982. Influence des Acides Amines Purlfes ajoutes dans les Aliments Composes, sur le Metabolisme des Pigments Carolenoides, chez Palaemon serratus (Crustacea, Decapoda). Aquaculture 28: 303–310.

Paradis, M.: Ackman, R.G. 1976. Localization of a Marine Source of Odd Chain-Length Fatty Acids. 1. The Amphipod Pontoporeia Femorata (Kroyer). Lipids 11(12): 863– 870.

Paradis, M.: Ackman, R.G. 1976. Localization of a Source of Marine Odd Chain-Length Fally Acids. II Seasonal Propagation of Odd Chain-Length Monoethylenic Fatty Acids in a Marine Food Chain. Lipids 11(12): 871– 876.

Patrois, J.: Ceccaldi, H.J.: Ando, T.: Kanazawa, A.: Teshima, S. 1978. Variation in Lipid Synthesis from Acetate during the Molting Cycle of Prawn. Bull. Jpn. Soc. Sci. Fish. 44(2): 139–141.

Petriella, A.M.: Muller, M.I.: Fenuccl, J.L. 1984. Influence of Dietary Fatty Acids and Cholesterol on the Growth and survival of The Argenline Prawn, Artemisa longinaris Bale. Aquaculture 37(1): 11–20.

Pledad-Pascual, F. 1984. Lecithin Requirement of Penaeus monodon Juveniles. First International Conference on the Culture of Penaeld Prawns/Shrimp, Ifoilo, Philippines. pp. 000

Pledad-Pascual, F. 1986. Effect of Supplemental Lecithin and Lipid Sources on the Growth and Survival of Penaeus monodon. Juveniles. In: Proc. of the First Asian Fisheries Forum, Manila, Phillppines, 26–31 May 1986. Maclean, J.L., L.B. Dizon and L.V. Hosillos (Eds) pp. 615–618.

Ponat, A.: Adelung, D.,1980. Studies to Establish an Optimal Diet for Carcinus maenas. II. Protein and Lipid Requirements. Mar. Biol. 60: 115–122.

Ponat, A. and Adelung. D., 1983. 1983. Studies to Establish an Optimal Diet for Carcinus maenas. III. Vitamin and Quantitative Lipid Requirements. Mar. Biol. 74: 275–279.

Provasoll, L.: Conklin, D.E.; D'Agostino, A.S. 1970. Factors Inducing Fertility in Aseptic Crustacea. Helgolander will. Meersunters. 20: 443–454.

Queiroz, J.F.; Leger, P.; Sorgeloos, P. 1989. The Effect of Broodstock Diet on Reproduction Activity and Offspring Quality In the Marine Crustacean Mysidopsis bahia (M). Abstracts of Second Asian Fisheries Forum OC012 pp. 66.

Ramos. T.L. Femandez, L.I. 1985. Quantitative Variations of Proteinemia and Total Ovarian Lipids During Gonadal Maturation of the Shrimp Penaeus notialis. Rev. Invest. Mar. 5(2).: 49–56.

Read, G.H.L. 1981. The Response of Penaeus indicus (Crustacea: Penaeidea) to Purified and Compounded Diets of Varying Fatty Acid Composition. Aquaculture 24: 245–256.

Reigh, R.C.: Slickney, R.R. 1989. Effects of Purified Dietary Fatty Acids on the Fatty Acid Composition of Freshwater Shrimp, Macrobrachium rosenbergil Aquaculture 77(⅔): 157–174.

Reinhardf, S.B.; Van Vteel, E.S. 1986. Lipid Composition of Twenty-Two Species of Antartic Midwater Zooplankton and Fish Mar. Biol. 91(1): 149–159.

Sandifer, P.A.: Joseph, J.D. 1976. Growth Responses and Fatty Acids Composition of Juvenile Prawns (Macrobrachium rosenbergli) Fed a Prepared Ration Augmented with Shrimp Head Oil. Aquaculture 8: 129–138.

Sarojini, R.; Jahagirdar, V.G. 1983. Role of SH-Linked Proteins and Saturated Lipids on the Ovarlan Maturation of the Hermit Crab, Pagurus kulkarniff. J. Adv. Zool. 42(2): 81– 85.

Shewbart, K.L.; Mies. W. 1973. Studies on Nutritional Requirements of Brown Shrimp. The Effect of Linolenic Acid on Growth of Penaeus aziecus. Proc. Wrold Mariculf. Soc. 4: 277–287.

Shudo, K.: Nakamura, K.: Ishikawa, S.: Kitabayashi, K. 1971. Studies on Formulated Feeds for Kurumia Prawn -IV. On the Growth-Promoting Effects of Both Squid Liver Oil and Cholesterol. Bull. Tokal Reg. Fish. Res. Lab., Tokyo 85: 129–138.

Sick, L.V.: Andrews, J.W. 1974. The Effect of Selected Dietary Lipids. Carbohydrates and Proteins on the Growth, Survival and Body Composition of Penaeus duorarum. Proc. World Maricutt. Soc. 4: 263–276.

Takada, H.; Kotani, S.: Tanaka, S.: Ogawa, T.: Takahashi, I.: Tsujimoto, M.: Komuro, T.: Shiba, T.: Kusumolo, S. et. al. 1988. Structural Requirements of Lipids a Species In Activation of Clotting Enzymes from the Horseshoe Crab, and the Human Complement Cascade Eur. J. Biochem. 175(3): 573– 580.

Takada, K.: Aokl, T.: Kunisaki, N. 1988. Proximate Composition, Free Amino Acid. Falty Acid, Mineral and Cholesterol Contents in Imported Frozen Shrimp. Bult. Jpn. Soc. Sci. Fish. 54(12); 2173–2179.

Takahashi, H.: Yamada. M. 1976. Lipid Composition of Seven Species of Crusiacean Plankton. Bult. Jpn. Soc. Sci. Fish. 42(7): 769–776.

Takeuchi, T.: Ackmari, R.G. 1987. Fatty Acid Composition of Triglycorldes of Rock Crab Cancer Irroratus Lipid. Bult. Jpn. Soc. Sci. Fish. 53(12): 2249–2253.

Teshima, S. 1971. In Vivo Transformation of Ergosterol to Cholesterol In Crab, Portunus trituberculatus. Bull. Jpn. Soc. Sci. Fish. 37: 671–674.

Teshima, S. 1971. Bioconversion of B-Silosterol and 24-Methylcholesterol to Cholesterol In Marine Crusiaceans. Comp. Biochem. Physiol. 39B: 815–822.

Teshima, S. 1972. Studies on the Sterol Metabolism In Marine Crustacean. Mem Fac. Fish., Kagoshima Univ. 21(2): 69–147

Teshima, S.: Ceccaidi, H.J.; Patrois, J.: Kanazawa, A. 1975. Bioconversion of Desmosterol to Cholesterol at Various stages of Molting Cycle in Palaemon serratus Pennani, Crustacea Decapoda. Comp. Biochem. Physiol. 50B: 485–489.

Teshima, S.; Kanazawa, A. 1970. Production of 11-Kelotestosterone and Other Sterols by the Sliced Ovaries of Crab, Portunus trituberculatus. Bull. Jpn. Soc. Sci. Fish. 36: 246–249.

Teshima, S.; Kanazawa, A. 1971. Sterol Composition of Marine Crustaceans. Bull. Jpn. Soc. Sci. Fish. 37: 63-

Teshima, S.: Kanazawa, A. 1971. In Vitro Bioconversion of Progesterone to 17alpha-Hydroxyprogesterone and Testosteronse by the Sliced Testes of Crab Portunus trituberculatus. Bull. Jpn. Soc. Sci. Fish 37: 524–528.

Teshima. S.: Kanazawa. A. 1971. Utilazation and Biosynthesis of Slerols in Artemia salina. Bull. Jpn. Soc. Sci. Fish. 37: 720– 723.

Teshima. S.: Kanazawa. A. 1971. Biosynthesis of Sterols in the lobster, Panulirus japonicus, the prawn Penaeus japonicus and the crab Portunus trituberculatus. Comp. Biochem. Physiol. 38B: 597–602.

Teshima, S.: Kanazawa, A. 1971. Bioconversion of the Dietary Ergosterol to Cholesterol In Artemia Salina. Comp. Biochem. Physiol. 38B: 603–607.

Teshima, S.: Kanazawa, A. 1971. Bioconversion of Progesterone by the Ovarles of Crab, Portunus Triturberculatus. General. Comp. Endocrinol. 17: 152–157.

Teshima, S.: Kanazawa, A. 1972. In Vivo Bioconversion of B-Sittosterol to Cholesterol In the Crab Portunus Triturberculatus. Mem. Fac. Fish., Kagoshima Univ. 21: 91– 95.

Teshima, S.: Kanazawa, A. 1973. Metabolism of Desmosterol in the Prawn, Penaeus Japonicus. Mem. Fac. Fish., Kagoshima Univ. 22(1): 15–19.

Teshima. S.: Kanazawa, A. 1976. Variation In Lipid Classes During the Molting Cycle of a Shrimp. Bull. Jpn. Soc. Sci. Fish. 42(10): 1129–1135.

Teshima, S.: Kanazawa. A. 1976. Comparison of the Sterol-Synthesizing Ability in Some Marine Invertebrates. Mem.

Teshima, S.: Kanazawa, A. 1978. Hemolymph Lipids of the Prawn - Short Paper Bull. Jpn. Soc. Sci. Fish. 44(8): 925-

Teshima, S.: Kanazawa, A. 1978. Release and Transport of Lipids in the Prawn. Bull. Jpn. Soc. Sci. Fish. 44(11): 1269–1274.

Teshima, S.: Kanazawa, A. 1979. Lipid Transport Mechanism in the Prawn. Bull. Jpn. Soc. Sci. Fish. 45(10): 1341–1346.

Teshima, S.: Kanazawa, A. 1980. Transport of Dietary Lipids and Role of Serum Lipoproteins in the Prawn. Bull. Jpn. Soc. Sci. Fish. 46: 151–55.

Teshima, S.: Kanazawa, A. 1980. Lipid consituents of Serum Lipoproteins in the Prawn. Bull. Jpn. Soc. Sci. Fish. 46(1):57– 62.

Teshima, S.: Kanazawa, A. 1980. Lipid Transport in Cruslaceans (Review). Min. Rev. Data File Fish. Res. 1:1–25.

Teshima, S.: Kanazawa, A. 1983. Variatiori in Lipid Composition During the Ovarian Maturation of the Prawn. Bull. Jpn. Soc. Sci. Fish. 49: 957–962.

Teshima, S.: Kanazawa, A. 1983. Digestibility of Dietary Lipids in the Prawn. Bull. Jpn. Soc. Sci. Fish. 49(6): 963–966.

Teshima, S.: Kanazawa, A. 1986. Nutritive Value of Sterols for the Juvenile Prawn. Bull. Jpn. Soc. Sci. Fish. 52: 1417–1422.

Teshima, S.: Kanazawa, A. 1988. Turnover of Dietary Cholesterol and Beta-Sitosterol in the Prawn. Bull. Jpn. Soc. Sci. Fish. 53: 601–607.

Teshima, S.: Kanazawa, A.: Horinouchi, K.; Koshio, S. 1988. Lipid Metabolism in Destalked Prawn, Penaeus japonicus. Induced Maturation and Transfer of Lipid Reserves to the Ovaries Bull. Jpn. Soc. Sci. Fish. 54: 1123–1129.

Teshima, S.: Kanazawa, A.; Horinouchi, K.: Yamasaki, S.: Hirata. H. 1988. Phospholipids of the Roller, Prawn and Larval Fish Bull. Jpn. Soc. Sci. Fish. 53: 605–615.

Teshima, S.: Kanazawa, A.: Kakuta, Y. 1986. Effects of Dietary Phospholipids on Growth and Body Composition of the Juvenile Prawn. Bull. Jpn. Soc. Sci. Fish. 52(1): 155–158.

Teshima S.: Kanazawa, A.: Kakula. Y. 1986. Effects of Dietary Phospholipids on Lipid Transport in the Juvenile Prawn. Bull. Jpn. Soc. Sci. Fish. 52(1): 159–163.

Teshima, S.: Kanazawa, A.: Kakuta, Y. 1986. Role of Dietary Phospholipids in the Transport of Radioactive Tripalmlin in the Prawn. Bull. Jpn. Soc. Sci. Fish. 52(3): 519–524.

Teshima. S.: Kanazawa, A.: Kakuta, Y. 1986. Role of Dietary Phospholipids in the Transport of [¹⁴ C] Cholesterol in the Prawn. Bull. Jpn. Soc. Sci. Fish. 52(3): 719–723.

Teshima, S.; Kanazawa, A.; Koshio, S.; Horinouchi, K. 1988. Lipid Metabolism in Destalked Prawn, Penaeus japonicus. Induced Maturation and Acclimation of Lipids in the Ovarles. Bull. Jpn. Soc. Sci. Fish. 54: 1115–1122.

Teshima, S.; Kanazawa, A.; Koshio, S.; Kondo, N. 1989. Nutritive Value of Sitosterol for the Prawn Penaeus Japonicus. Bull. Jpn. Soc. Sci. Fish. 55(1): 153–157.

Teshima, S.; Kanazawa, A.; Okamoto, H. 1974. Absorption of Sterols and Cholesteryl Esters in a Prawn, Penaeus japonicus. Bull. Jpn. Soc. Sci. Fish. 40(10): 1015–1019.

Teshima, S.; Kanazawa, A.: Okamoto. H. 1976. Sterol Biosynthesis from Acelate and the Fate of Dietary Cholesterol and Desmosterol in Crabs, Bull. Jpn. Soc. Sci. Fish. 42(11): 1273–1280.

Teshima, S.; Kanazawa, A.; Okamoto, H. 1976. Analysis of Falty Acids of Some Crustaceans. Mem. Fac. Fish., Kagoshima Univ. 25(1): 41–46.

Teshima, S.; Kanazawa, A.; Okamoto, H. 1977. Variation in Lipid Classes During the Molting Cycle of the Prawn Penaeus japonicus. Mar. Biol. 39: 129–136.

Teshima, S-I.; Kanazawa, A. 1982. Variation In Lipid Compositions during the Larval Developement of the Prawn (Penaeus japonicus). Mem. Fac. Fish. Kagoshima U. 31: 205–212.

Teshima, S-l; Kanazawa, A.; Kakuta, Y. 1986. Growth, Survival and Body Lipid Composition of the Prawn Larvae Receiving Several Dietary Phospholipids. Mem. Fac. Fish. Kagoshima Univ. 35(1): 17–27.

Teshima, S-l; Kanazawa, A.; Koshio, 8.; Horinouchi, K. 1989. Lipid Metabolism of the Prawn. Penaeus japonicus During Maturation: Variation in Lipid Protlies of the Ovary and Hepalopancreas. Comp. Biochem. Physiol. 92B(1): 45–50.

Teshima, S-l; Kanazawa, A; Sasada, H. 1983. Nutritional Value of Dislary Cholesterol and other Sterols to Larval Prawn, Penaeus japonicus Bale. Aquaculture 31: 159–167.

Teshima, S-l.; Kanazawa, A.; Sasada, H.; Kawasaki, M. 1982. Requirements of the Larval Prawn, Penaeus japonicus for Cholesterol and Soybean Phospholipid. Mem. Fac. Fish., Kagoshima U. 31:193–199.

van den Oord, A. 1964. The Absence of Cholesterol Synthesis in the Crab, Cancer pagurus L. Comp. Biochem. Physiol. 13: 461–467.

van det Horst, D.J.: Oudejans, R.C.H.M.; Plug, A.G.; van der Sluls, I. 1973. Fatty Acids of the Female Horseshoe Crab Xiphosura (Limulus) Polyphemus. Mar. Biol. 20:291– 296.

Villegas, C.T.; Kanazawa, A. 1978. Relationship Between Diet Compostion and Growth Rate of the Zoeal and Mysis Stages of Penaeus japonicus Bale. Quart. Res. Req. Aquacult. Dept. Southeast Asia Fisheries Dev. Centre 2(2): 24–29.

Wallace, R.A.; Walker, S.L.; Hauschka, P.V. 1967. Crustacean Lipovitellin Isolation and Characterization of Major High-Density Lipoprotein from the Eggs of Decapods. Biochemistry 6: 1582–1590.

Ward, D.G.: Middleditch, B.S.; Missier, S.R.; Lawrence, A.L. 1979. Fatty Acid Changes During Larval Development. Proc. World. Maricull. Soc. 10: 464–471.

Watanabe, T.; Oowa, F.; Kllajima, C.; Fujila, S. 1978. Nutritional Quality of Brine Shrimp, Artemia Safina, as a Living from the Viewpoint of Essential Fatty Acid for Fish. Bull. Jpn. Soc. Sci. Fish. 44(10) : 1115– 1121.

Whitney, J.O. 1970. Absence of Sterol Biosynthesis in the Blue Crab Callinectes sapidus Rathbun and in the Barnacle Balanus nubilus Darwin. J. Exp. Mar. Biol. Ecol. 4: 229–237.

Wolfe, D.A.; Rao, P.V.; Cornwell, D.G. 1965 Studies on the Fatty Acid Composition of Crayfish Lipids. J.Am. Oil Chem. Soc. 42: 633–637.

Zandee, D.I. 1967. Absence of Cholesterol Synthesis as Contrasted with the Presence of Fatty Acid Synthesis in some Arthropods. Comp. Biochem. Physiol. 20: 811–822. Zandee. D.T. 1962. Lipid Metabolism in

Annex K
Crustacean Protein and Amino Acid Requirements

By

Dr. John D. Castell
Dept. of Fisheries and Oceans, Biological Sciences Branch,
Benthic Fisheries and Aquaculture Division, Scotia-Fundy Region,
P.O. Box 550, Hallfax, Nova Scotia B3J 2S7, Canada.

Alava, V.R.: Lim, C. 1983. The Quantitative Dietary Protein Requirements of Penaeus monodon Juveniles in a controlled Environment. Aquaculture 30:53–61.

Ali, S.A. 1982. Relative Effeciencies of Pelleted Feeds Compounded with Different Animal Proteins and the Effect of Protein Level on the Growth of the Prawn, Penaeus indicus. Symp. Ser. Mar. Biol. Assoc. India. 6: 321–328

Andrews, J.W.; Sick, L.V.: Baptist, G.J. 1972. The Influence of Dietary Protein and Energy Levels on Growth and Survival of Penaeid Shrimp. Aquaculture 1:341–347.

Antiporda, J.L. 1986. Optimum Dietary Protein Requirement for Macrobrachium rosenbergii juveniles. Bankok (Thailand), NACA pp. 20.

Bages, M.; Sloane, L. 1981. Effects of Dietary Protein and Starch Levels on Growth and Survival of Penaeus monodon (Fabricius) Postlarvae. Aquaculture 25: 117–128.

Balazs, G.H.; Ross, E. 1976. Effect of Protein Source and Level on Growth and Performance of the Capitive Freshwater Prawn, Macrobrachium rosenbergi. Aquaculture 7:299– 313.

Bhaskar, T.I.C.J.; All, S.A. 1984. Studies on the Protein Requirement of Postlarvae ot the Penaeld Prawn Penaeus indicus H. Milne Edwards Using Purified Diets. Indian J. Fish. 31(1): 74–81.

Boonyaratpalin, M.; New, M.B. 1980. Evaluation of Diets for Giant Prawn Reared in Concrete Ponds. Thai Fish. Gaz. 33: 555–561.

Boonyaratpalin, M.; New, M.B. 1982. Evaluation of Diets for Macrobrachium rosenbergil Reared in Concrete Ponds. In: Glant Prawn Farming. M.B. New (Ed.) pp. 249–256.

Campillo, A.; Luquet, P. 1976. Effect of Protein Level on the Growth of Palaemon serratus (Pennant) Raised in the Laboratory. Rev. Trav. Inst. Peches marit. 39:407–414.

Castell, J.D.: Budson, S.D. 1974. Lobster Nutrition: The Effect on Homarus americanus of Dietary Protein Level. Journal of the Fisheries Research Board of Canada 31:1363–1370.

Castell, John, D.: Kean, Joan C.: D'Abramo, Louis R.: Conklin, Douglas E. 1989. A Standard Reference Diet for Crustacean Nutrition Research. I. Evaluation of Two Formulation. J. World Aquacult. Soc. 20(3); 93–99.

Castell, John D.; Kean, Joan C.: McCann, D.G.C; Boghen, Andrew D.: Conklin, Douglas E; D'Abramo, Louis R. 1989. A Standard Reference Diet for Crustacean Nutrition Research. II. Selection of a Purification Procedure for Production of the Rock Crab Cancer irroratus Protein Ingredient. J. World Aquacult. Soc. 20(3):100–106.

Celada, J.D.; Carral, J.M.; Gaudioso, V.R.; Termino, C.: Fernandez. R. 1989. Response of Juvenile Freshwater Crayfish (Pacifiasticus leniusculus Dana) to Several Fresj and Artificially Compounded Diets. Aquaculture. 76: 67–78.

Clifford, H.C., II: Brick, R.W. 1978. Protein Utilization in The Freshwater Shrimp, Macrobrachium rosendergil. Proc. World Maricult. Soc. 9: 195–208.

Colvin, P.M. 1976. Nutritional Studies on Penaeld Prawns: Protein Requirements in Compounded Diets for Juvenile Penaeus indicus (Milne Edwards). Aquaculture 7:315–326.

Cowey, C.B.: Forster, J.R.M. 1971. The Essential Amino-Acid Requirements of the Prawn Palaemon serratus. The Growth of Prawn on Diets Containing Proteins of Different Amino-Acid Compositions. Mar. Biol. 10:77–81.

Cruz-Ricque, L.E.: Gullaume, J.; Cuzon, G. 1987. Squid Protein Effect on Growth of Four Penaeld Shrimp. J. World Aquacult. Soc. 18: 209–217.

Cruz-Ricque, L.: Guillaume, J.: van Wormhoudt, A. 1989. Effect of Squid Extracts on Time Course Appearance of Glucose and Free Amino Acids in Haemolymph in Penaeus japonicus after Feeding: Preliminary Results. Aquaculture 76: 57–65.

Cruz-Suarez, L.E.; Guillaume, J.; Van Wormhoudt, A. 1987. Effect of Various Levels of Squid Protein on Growth and Some Biochemical Parameters of Penaeus Japonicus Juveniles. Bull. Jpn. Soc. Sci. Fish. 53: 2083–2088.

Dall, W.; Smith, D.M. 1986. Oxygen Consumption and Ammonia-N Excretion In Fed and Starved Tiger Prawns, Penaeus esculentus Haswell. Aquaculture 55: 23–33.

Depledge, M.H.; Bjerregaard, P. 1989. Hemolymph Protein Composition and Copper Levels in Decapod Crustaceans. Helgol. Meeresunters 43(2): 207–224.

Deshimaru, O. 1976. Studies on a Purified Diet for Prawn. VIII. Changes in Free Amino Acid Content in Muscle, Hepatopancreas and Blood of Prawn after Feeding. Bull. Jpn. Soc. Sci. Fish. 42(5): 655–660.

Deshimaru, O.; Kuroki, K. 1975. Studies on a Purified Diet for Prawn. IV. Evaluation of Protein, Free Amino Acids and Their Mixture as Nitrogen Source. Bull. Jpn. Soc. Sci. Fish. 41: 101–103.

Deshimaru, O.; Kuroki, K. 1975. Studies on a Purified Diet for Prawn. V. Evaluation of Casein Hydrolyzates as a Nitrogen Source. Bull. Jpn. Soc. Sci. Fish. 41(3): 301–304.

Deshimaru, O.; Yone, Y. 1978. Optimum Level of Dietary Protein for Prawn. Bull. Jpn. Soc. Sci. Fish. 44: 1395–1397.

Dy Penaflorids, Veronica. 1989. An Evaluation of Indigeneous Protein Sources as Potential Component in the Diet Formulation for Tiger Prawn, Penaeus monodon, Using Essential Amino Acid Index (EAAI). Aquaculture 83(3/4):319–330.

El-Dakor, S. 1986. Effects of Different Dietary Protein: Energy Rations on the Growth and Survival of Penaeus semisulcatus Dh Haan (Decapoda: Penaeidae). Kuwait Bull. Mar. Sci. 0: 213–222.

Fair, P.H.; Sick, L.V. 1982. Serum Amino Acid Concentrations During Starvation in the Prawn, Macrobrachium rosenbergii, as an Indicator of Metabolic Requirements. Comp. Biochem. Physiol. 73B: 195–200.

Farmanfarmarian, A.; Lauterio, T. 1979. Amino Acid Supplementation of Feed Pellets of the Giant Shrimp (Macrobrachium rosenbergil). Proc. World Maricult. Sco. 10: 674–688.

Famanlarmarian. A.: Lauterio. T. 1988. Amino Acid Composition of the Tail Muscle of Macrobrachium rosenbergli. Comparison to Amino Acid Patterns of Supplemented Commercial Feed Pellets. Proc. World Maricult. Soc. 11: 454–462.

Fenucci, J.L: Zein-Eldin, Z.P.: Lawrence, A.L. 1980. The Nutritional Responses of Two Penaeid Species to Various Levels of Squid Meal in a Prepared Feed. Proc. World Maricult. Soc. 11: 403–409.

Forrellat, A.; Gonzalez, R.; Carrillo, O. 1988. Evaluation of the Protein Quality of Food for Shrimp. Rev. Invest. Mar. 9(1): 71–80.

Fruechtenicht, G.W. 1988. The Effect of Protein Level in Isocaloric Feeds on the Growth Performance of Macrobrachium rosenbergii Individually Reared in Clear Water Flow-through Aquaria. Pac. Sci. 42(1–2): 119–120.

Gagné, G.; Boghen, A.D.; Castell, J.D. 1986. Influence d'une supplémentation en acides aminés dans un régime à base de protèines de crabe chez des homards juvéniles (Homarus americans). Reprod. Nutr. Dévelop. 26: 1265– 1272.

Gómez D., G., H. Nakagawa and S. Kasahara, 1988. Effect of Dietary Protein/Starch Ratio and Energy Level on Growth of the Giant Freshwater Prawn Macrobrachium rosenbergii. Bull Jpn Soc. Sci Fish 54: 1401–1407.

Hajra, A.; Ghosh, A.; Mandal, S.K. 1988. Biochemical Studies on the Determination of Optimum Dietary Protein to Energy Ratio for Tiger Prawn, Penaeus monodon (Fab.), Juveniles. Aquaculture 71(½): 71–79.

He, H. 1988. A Study on the Essential Amino Acids of the Prawn Penaeus orientalis. Oceanol. Limnol. Sin. 19(4): 307–313.

He, Haiqi 1988. Essential Amino Acids of the Prawn Penaeus orientalis. Haiyang Yu Huzhao 19(4): 307–313.

Hubbard, D.M.; Robinson, E.H.; Brown, P.B.; Daniels, W.H. 1986. Optimum Ratio of Dietary Protein to Energy for Red Crayfish (Procambarus clarkii). Prog. Fish-Cult. 48(4): 233–237.

Huner, J.V.; Meyers, S.P. 1978. Dietary Protein Requirements of Red Swamp Crawfish. Procambarus clarkii (Girard) (Decapod; Cambaridae). Assoc. Soputheast Biol. Bull 25(2): 70-?.

Huner, J.V.; Meyers, S.P. 1979. Dietary Protein Requirements of Red Swamp Crawfish. Procambarus clarkii (Girard) (Decapod; Cambaridae), Grown in a Closed System. Proc. World Maricult Soc. 10: 751–760.

Kanazawa, Akio; Teshima, Shin-ichi. 1981. Essential Amino Acids of the Prawn. Bull. Jpn. Soc. Sci. Fish. 47: 1375–1377.

Kanazawa, Akio; Teshima, Shin-ichi; Matsumoto, Seiki; Nomra, Tadacuna. 1981. Dietary Protein Requirement of the Shrimp Matapenaeus monoceros. Bull. Jpn. Soc. Sci. Fish. 47(10): 1371–1374.

Katzen, S.; Salser. B.R.; Ure, J. 1984. Dietary Lysine Effects on Stress-Related Mortality of the Marine Shrimp, Penaeus stylirostris. Aquaculture 40: 277–281.

King, F.D.; Cucci, T.L.; Bidigare, R.R. 1985. A Pathway of Nitrogen Metabolism in Marine Decapod Crabs. Comp. Biochem. Physiol. 80B: 401–403.

Kitabyashi, K.; Shudo, K.; Nakamura, K.; Ishikawa, S. 1971. Studies on Formula Feeds for Kuruma Prawn - III. On the Growth-Promoting Effects of both Arginine and Methionine. Bull. Tokai Reg. Fish. Res. Lab., Tokyo. No. 65: 119–128.

Kitabyashi, K.; Shudo, K.; Nakamura, K.; Ishikawa, S. 1971. Studies on Formula Feeds for Kuruma Prawn - V. On the Growth-promoting Effects of Protein Level in a Diet and Fre-examination of Ingredients used. Bull. Tokai Reg. Fish. Res. Lan No. 65: 139–147.

Koshio, S.; Kanazawa, A.; Teshima, S.-I.; Castell, J.D. 1989. Nutritional Evaluation of Crab Protein for Larval Penaeus japonicus fed Microparticulate Diets. Aquaculture 81(2): 145– 154.

Lasser, G. 1975. Amino Acid Requirements of the Dunganess Crab. Proc Natl. Shellfish Ass. 65:9.

Liang Yaquan; Ji Wenjuan. 1986. Protien Requirement in Formulated Diets for Penaeid Shrimp (Peaneus orientalis) in Different Growth Stage. Mar. Fish. Res., (Chinese) 7: 79–87.

Lim, G.; Suraniranat, P.: Platon, R. 1978. A Preliminary Study on the Elevation of Casein, Shrimp Meal, Squid Meal and Spirulina as Protein Sources for Penaeus monodon (Fabricus) Postlarvae. Quart. Res. Rep. Southeast Asian Fish. Dev. Cent. Aquacult. Dept. 2(2): 13–18.

Mai, Kangsen; Li, Aijie; Yin, Zuofen. 1988. Studies on the Absorption and Utilization of Amino Acids in the Test Diets by the Prawn, Penaues orientalis. Acta Oceanol. Sin/Haiyang Xuebao. 7(4): 621–629.

Mai, Kangsen: Li, Aijie; Yin, Zuofen. 1987. Absorption and Utilization of Proteins and Amino Acids from Feeds by Penaeus orientalis. Acta Oceanol. Sin/Haiyang Xuebao (Zhongwenban) 9: 489–495.

Marangos, C.; Ramos, L.; Oliva, M. 1989. Variations of Free amino Acid Levels in the Ovary, Hepatopancreas and Hemolymph of Penaeus Schmitti (Crustacea, Decapoda, Penaeidae) During Ovarian Maturation. Arch. Int. Physiol. Biochem. 97(1): 95–106.

Millikin, M.R., A.R. Fortner, P.H. Fair and L.V. Sick, 1980. Influence of Dietary Protein Concentration on Growth, Effd Conversion and General Metabolism of Juvenile Prawn (Macrobrachium rosenbergil). Proc. World Maricult. Soc. 11:382–391.

Millikin, M.R., G.N. Biddle, T.C. Siewicki, A.R. Fortner and P.H. Fair, 1980. Effects of Various Levels of Dietary Protein on Survival, Molting Frequency and Growth of Juvenile Blue Crabs (Callinectes sapidus). Aquaculture 19:149–161.

Moureau, C.E.; Ceccaldi, H.J. 1985. Variations Circadiennes des Acides Amines Libres de l'Hemolymphe de Penaeus japonicus. Biochem. System. Ecology. 13(1): 35–40.

Norman-Boudreau, Karen; Conklin, Douglas E. 1985. Antinutritional Properties of Egg White in Diets for Lobsters. Bull. Jpn. Soc. Sci. Fish. 51(6):953–957.

Otazu-Abrill, M.; Martin, B.J.; Ceccaldi, H.J. 1982. Influence of Purified Amino Acids Added to Artificial Diets on the Metabolism of Carotenoid Pigments in Palaemon serratus. Aquaculture. 28(3–4): 303–310.

Prasad, P.N.: Neelakantan, B. 1986. Proximate and Essential Amino Acid Composition in Edible Crab, Scylla serrata. Comp. Physiol. Ecol. 14(1):34– 37.

Sambasivam, S.: Subramanian, P.: Krishnamurthy, K. 1982. Observations on Growth and Conversion Efficiency in the Prawn Penaeus indicus (H. Milne Edwards) Fed on Different Protein Levels. Symp. Ser. Mar. Biol. Assoc. India. 6: 406–409.

Sedgwick, R.W., 1979. Influence of Dietary Protein and Energy on Growth, Food Consumption and Food Conversion Efficiency in Penaeus merguiensis De Man. Aquaculture 16:7–30.

Shewbart, K.L.: Mies, W.L.: Ludwig, P.D. 1972. Identification and Quantitative Analysis of the Amino Acids Present in Protien of the Brown Shrimp Penaeus aztecus. Mar. Biol. 16: 64–67.

Shigeno, K.,K. Kumada. O. Deshimaru, T. Aramaki, K. Kuroki and K. Kitaue, 1972. Studies on the Artificial Diets of Prawn - 1. Relationship Between the Feed Efficiency and Crude Protein in the Diets. Bull. Jpn. Soc. Sci. Fish. 38:101–106.

Smith, Linda L.: Lee, Phillip G.: Lawrence, Addison L; Strawn, Kirk. 1985. Growth and Digestibility by Three Sizes of Penaeus vannamei Boone: Effects of Dietary Protein and Prltein Source. Aquaculture. 46(1):85–96

Su, Y.; Xiao. J. 1987. Studies on the Essential Amino Acids of two Marine Planktonic Crustaceans. J. Xiamen Univ. 26(4):503–508.

Su, Y.: Xiao, J. 1987. Studies on the Essential Amino Acids of Two Marine Planktonic Crustaceans. Xiamen Daxue Xuebao. Ziran Kexueban (Chinese) 26:503–508.

Teles, A.O. 1985. Protein Requirements of Palaemon Elegans. Publ. Inst. Zool. Dr. Augusto Nobre. No 189.

Teshima, S-I.; Kanazawa, A.: Yamashita. M. 1986. Dietary Value of Several Proteins and Supplemental Amino Acids for Larvae of the Prawn Penaeus Japonicus. Aquaculture 51: 225–235.

Van Wormhoudt, A.; Cruz, E; Gullaume, J.; Favrel, P. 1986. Effect of the Soybean-Tripsin Inhibitor on Growth and Digestive Enzymes in Penaeus japonicus (Crustacea: Decapoda): Possible Role of Gastointestinal Peptides. Océanis 12: 305– 319.

Venkataramiah, A., G.J. Lakshmi and G. Gunter, 1975. Effect of Protein Level and Vegetable Matter on Growth and Food Concersion Efficiency of Brown Shrimp. Aquaculture 6:115–125.