The H 440 standard reference diet of Halver (1969) has*

proven satisfactory for use with a variety of species of fishes.

If this exact formula does not prove satisfactory for growth and

survival of the test fish, slight modifications of clearly

explained ingredient changes still permit meaningful comparisons

of the test fish results with other species

Ahamed Ali, S

Kanazawa, Akio

Paulraj, R

CMFRI Digital Repository

^PM^BlQMfflM^SMQMQM^HiQH •/' f r \ Phe ^ His ' f'^ , • % -i0P? ': i*P P^ ie* « COOH ^4& k i ^ <|A jg : m «? up. Ey^^ftni^^fflB^ ^ B^"rijiBMSSMt^ Br QlTiJ^B I^B •• >'Vi :'f{ij;j..V^;. i>_V'" ^ ..--.* CMFRI SPECIAL PUBLICATION Number 8 MANUAL OF RESEARCH METHODS FOR FISH AND SHELLFISH NUTRITION Issued on the occasion of the Workshop on METHODOLOGY FOR FISH AND SHELLFISH NUTRITION organised by The Centre of Advanced Studies in Mariculture, Central Marine Fisheries Research Institute, held at Cochin from 11 -16 January 1982 Published by: E. G. SILAS Director Central Marine Fisheries Research Institute COCHIN " " " " • • • f f c f PREFACE The Centre of Advanced Studies in Mariculture established at the Central Marine Fisheries Research Institute has been conducting '.Jorkshops in Research Methodologies on specialised disciplines with a view to enhance the competence of the scienti-fic workers specialising in researches connected with mariculture. The main emphasis in mariculture research has been directed towards the development of economically viable culture techniques for culturable species of fish and shellfish, with a view to augmenting the fish and shellfish production of the country. In order to develop low-cost technologies the essential operational inputs have to be rationally utilized. + It has been well established that feeding constitutes the major cost of production, often exceeding 50 per cent of the » operating costs in intensive aquaculture operations. Two main factors affecting the cost of feeding are composition of the diet and efficiency of feed conversion. In order to develop least-cost formula diets of high conversion efficiency, knowledge of the nutritional requirements of the different species during the different phases of the life cycle and the nutritive value of the complex feed ingredients available in the country to the candidate species is a prerequisite. The existing information on the nutritional requirements of cultivated species of fish and shellfish in India, is meagre and recently research has been intensified in this area. If researches on this field could be carried ut using standardised experimental procedures, the data obtained on the nutritional requirements ot the different species could be stored in a fish and shellfish nutrition data batik, from where data could be disseminated to the users such as feed manufacturers, farmers, extension workers and research workers as and when required. It. is also necessary that the data collected on the chemical composition of the feed ingredients and their nutritive value tor the species should be based un standard chemical methods and experimental procedures so that the data could be stored in ii the dafca batik which eventually could become a National Fish Seed information. yCentpe.. i , r .'i;.u .,To.iUBd?fftake studies on the above lines, especially by the technicians and .research workers entering afresh into the field/ the,need of praptiqal guides describing the research techniques and methods, planning of investigations, collection of data and their interpretation peed not be emphasized. Keeping this in Y£»w<,.;the, .present manual, on Research Methods in Fish and Shellfish Nutrition is issued by the centre<of Advanced studies in Mari-culture on the occasion of the Workshop on Methodology of Fish and Shellfish Nutrition. Drf Akio Kanajawa, Professor of Nutritional Chemistry, University of Kagpsh,iniat| Japan and Consultant in Fish and Shell-fish Nutrition at the CAS in Mariculture, has been kind enough to cooperate with the scientists of CAS in Mariculture of the Central Marine Fisheries Research institute in the preparation of t£i» Bvanua,!. There are chapters in this manual covering various methods on(composition analysis of feeds, including growth inhibitors and toxins; deterrainetlpn of digestibility coefficient; protein evaluation; bioenergetics; determination of essential amino acid requirements using radioisotope method; research, test diets for fishes and prawns; feed formulation methods; experimental design, etc. Methods of preparation of mlcroparfciculate diets, phytoplankton and zppplankton culture methods, etc, are also included to facilitate larval nutrition studies. , Many of the methods given in the manual have been standardized for fish and shellfish nutrition studies in India and abroad. , The users can also gain maximum benefit by suitable modifications of other methods which are given as guidelines. I would like to thank ail the scientific and technical staff especially Shri s. Ahamed All, Dr. K. Alagarswami, Shri D.C.V. Ea'sterson, Shri C.P., Gopina^han, Shri T. Jacob, Shri M.S. Muthu,,Dr. R. Paul Raj, Dr. A.G. ponniah and ill / in:, p. vedavyasa Rao who have rendered assistance during the preparation of this manual. Thanks are also due to Shri Johnson, Librarian and shri Kambadkar, Technical Assistant, Central Marine Fisheries Research institute, for the help rendered bythtra in printing this manual. (E.G. Silas) Director, CMPRI, Sub-Project Coordinator, Centre of Advanced studies in Maricultuce CHAPTER 12 PREPARATION OF ARTIFICIAL DIETS FOR NUTRITIONAL STUDIES 1 TEST DIET FOR FISHES The H 440 standard reference diet of Halver (1969) has* proven satisfactory for use with a variety of species of fishes. If this exact formula does not prove satisfactory for growth and survival of the test fish, slight modifications of clearly explained ingredient changes still permit meaningful comparisons of the test fish results with other species. Diets may be prepared as muist, semi-moist or dry diet, or as a powder, rolled pellets, extruded pellets, or compressed pellets. 1.1 Standard Reference Diet H-440 Complete test diet Vitamin-free casein White dextrin Gelatin Corn oil Cod liver oil Vitamin mixture Mineral Mix Total Water Total diet as feed (g> 38 28 12 6 3 9 4 100 200 300 Vitamin Mixture L-cellulose Choline chloride inositol L-Ascorbic acid Nicotinic acid Ca-pantothenate Riboflavin Thiamine HCl Pyridoxine HCl Menadione (K) Folic acid Vitamin B., Biotin L-tocopherol-acetate (E) (9) 8.000 0.500 0.200 0.100 0.075 0.050 0.020 0.005 0.005 0.004 0.0015 0.0011 0.0005 0.040 University of Kagoshima, Japan, n. Paul Raj and 3yed Ahamed All, Central Marine Fisheries Research Institute, Cochin-18. ?1 Miner. ji ;:ix USP XII No.2 A1C13 6H,0 2n S.54.H2O cuci MnS04 KI coci2 H20 6H20 (g> ugp X I I MO.2 99.50 Calcium Diphosphate 0.015 Calcium lactate 0.300 Ferric citrate 0.010 Magnesium sulphate 0.080 Potassium phosphate (dibasic) 0.015 Sodium Diphosphate 0.100 Sodium chloride (9) 13.53 32.70 2.97 13.20 23.98 99.50 1.2 Preparation of Halver's diet Gelatin Take in the container, add cold water and heat with stirring in a water bath at 80°C to dissolve gelatin, remove from heat add with stirring - dextrin (first) casein (second) minerals., oils and vitamins as temperature decrease mix well until the temperature decreased to 40'C Warm diet transfer into another container put into the refrigerator at 5"C Artificial diet Mix L-tocopherol with oils Mix 1 part of vitamins with 8 parts of L-cellulose and Add vitamin B,2 in water during final mixing. 2 TEST DIET FOR THE PRAWN, PBHAEJS JAP0NICU3 Ingredients Casein (vitamin-free) Clucose Sucrose a 50.0 5.5 10.0 Ingredients £ a-Starcli Glucosamine nci Sodium citrate Sodium succinate Cholesterol Pollack liver oil Minerals*1 Vitamins*2 Cellulose powder + Water 130-135 ml 4.0 0.3 0.3 0.3 0.5 12.0 3.5 3.2 1.9 100.02 *1, *2 Minerals and vitamins for thejjtest diet of prawn, P. {aponlcus Vitamins Thiamine HC1 (Bx) Riboflavin (B2) P. Aminobenzoic acid Biotin Inositol Niacin Ca Panthothenate Pyrldoxlne HC1 22 Vitamins 4.9 Menadione 8;0 ^-Carotene 10.0 a^Tocopherol (vitamin E) 0.4 Caicipherol 00.0 Cyanocobalamine (B,,) 40.0 Na Ascorbate (vitamin C) 60.0 Folic acid 12.0 Choline HCl 4.0 9.6 20.0 1.2 0.08 2000.00 0.30 600.00 Total 32000 Minerals K2HP04 Ca3(P04)2 NaH2P042H20 MgS047H20 HnS045H20 FeS0.7H 0 2 2.0 2.72 0.79 3.02 0.004 0.015 Total 8.549 93 2.1 Preparation of Test Diet3 for the Prawn i) Vitamins are dissolved in either water or ethanol and stored in a refrigerator at S°C. ii) Minerals are powdered into a fine particle with a •iiortar and mixed well. iii) The pH of diet is adjusted with 0.5N-NaOH (in most case, 65-70 ml. of 0.5N-NaOH solution) to pH 6.3-7.0 (optimum pH for the prawn diet). Detailed procedures: All the ingredients are weighed and powdered into a fine particle. Casein, glucose, sucrose, a-starch, gluco-samine HC1, Na citrate. Ha succinate, cholesterol, and agar are combined and mixed thoroughly. To this, cellulose powder, minerals, vitamins, and pollack liver oil -ere added in turn and mixed well. Finally, distilled water (130-135 ml) is added to the mixture of dry ingredients and mixed again, the pH of diet being adjusted to pH 6.8-7.0 with 0.5N-NaOH (or 1N-HC1). The mixture so obtained is heated in autoclave (without addition of pressure) at 100°C for 10 min. The container is taken out from the autoclave, and the ingre-dients are mixed well and again heated at 100°C for 10 min, by the same manner. The heated mixture of the ingredients is packed into the plastic tubas (Cleharon tubes) by the similar manner to that ;f casing of sausage-making. The cased diet is again heated at 100°C for 10 min. and then cooled. 3 REF3SSMCES 1. Castell, J.D. Report of tho 2IFAC, rail 3 and ICES and K. Tiews working group on .-standardisation of 1930 methodology in fish nutrition research EIFAC Technical Papqr Wo. 36. 24 pp. t 2. Halver, J.2. (ed:), ?ish Nutrition. New York, Academic Press, 719 p. 3. Manual 3ooK of E;:porimant3, Laboratory of Fisheries Chemistry, University of Kagoshlma, Japan, 1930-31, 1 l 

Preparation of artificial diets for nutritional studies

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Preparation of artificial diets for nutritional studies

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