ANNEX A

PROJECT PROFORMA FOR NUTRITION AND DIET DEVELOPMENT PLANS

due date:

1. Add new major programme to NDD plans

   D.

   Digestibility coeficients for lipid/energy in feedstuffs

   01-06-89

   
   

2. Develop proforma for B. Area on protein digestibility

   B-1	Protein DC for 5 feedstuffs by Tilapia	done
   B-2	Protein DC for 5 feedstuffs by Rohu	done
   B-3	Protein DC for 5 feedstuffs by Mrigal	01-07-89
   B-4	Protein DC for 5 feedstuffs by Clarias	01-08-89

   
   

3. Develop proforma for C. Area on formulation and testing

   C-1	Development/testing of fry to fingerling feed for Rohu	01-06-89
   C-2	Development/testing of fry to fingerling feed for Puntius	15-06-89
   C-3	Development/testing of fry to fingerling feed for Clarias	15-06-89
   C-4	Development/testing of supplementary feed for pond production of Clarias	01-09-89
   C-5	Utilization of blood cockle meat in semi-moist diets for shrimp	01-08-89
   C-6	Supplementary feeds for pond rearing of common and major carps in standard polyculture techniques	01-09-89

   
   

4. Develop proforma for D. Area on digestibility coeficients of lipid and energy components of feedstuffs

   D-1	DC of lipid and energy components by Tilapia	01-05-90
   D-2	DC of lipid and energy components by Rohu	01-05-90
   D-3	DC of lipid and energy components by Mrigal	01-05-90
   D-4	DC of lipid and energy components by Clarias	01-05-90

ANNEX B

PLANS AND PROGRESS 1985 – 1989

(--- = Plans; = Progress; +++ = Deleted from Plan)

ANNEX C

BARC    
Form No. P-1

Bangladesh Agricultural Research Council
          RESEARCH PROPOSAL

Received at BARC
Date :
By :

Project title: D.DIGESTIBILITY STUDY OF LIPID AND ENERGY COMPONENTS OF DIFFERENT FEEDSTUFFS LEADING TO THE FORMULATION OF QUALITY FISH FEEDS FOR DIFFERENT CULTIVATABLE SPECIES.

Implementing Agency

A. Proposal content :-

1. Importance/significance of research work :

Successful aquaculture projects whether intensive or semi-intensive depends upon to a large extent on the feeding practice of suitable/balanced diets. In Bangladesh one of the constraints to increase fish production is the formulation and preparation of nutritionally balanced diets to meet the requirements of fish. Fortunately our country has the highest water : land ratio in the world. If properly managed and harvested the fish production of these water bodies would be increased and indeed solve part of our present malnutrition problem.

For the formulation of a quality fish feed we must know the nutrient content and quantity of the feedstuffs. In Bangladesh a large variety of indigenous raw materials and agro-based industrial by products rich in nutrients ultimately go to waste without being properly utilized many of these can be used as fish feed ingredients. The important one among them are : fish meal, blood meal, animal offal, kachuripana whole, plant, stem, leaves, flowers, topa pana, kudi pana, rice bran, wheat bran, soyagretes, masur bran, mung bran, slaughter house waste, kitchen waste, silkworm pupae, prawn shell, crab shell, fish silage, crab meal, etc. A comprehensive list of potential feed ingredients including their sources, season of availability, amount available, biochemical composition should be made at the national level, However, to know the detail information about their availability, economy, biochemi cal composition, FARS, Mymensingh has already started a programme to survey potential fish Feed. Ingredients ( FARS-FNI) which will make a systematic approach towards making a sound and effective national policy for the formulation and manufacture of fish feed from locally available ingredients. One major prerequisite of these organic wastes in feed formulation is information on the digestibility of feed ingredients to fish.

A feedstuff may appear from its chemical composition to be an excellent source of nutrients but will be of little actual value unless it can be digested and absorbed. Feedstuff digestibility is most often expressed in terms of digestiole energy (DE), which is defined as the energy in the food comsumed, or intake energy (IE), minus the energy present in the feces. (FE) :

DE = IE - FE

Lovell ( 1977 ) indicated that nutrient digestibility among feedstuffs varied considerably for different warm water fishes. Knowledge of nutrient availability from the various feedstuffs used in formulating warm water fish diets is desirable so that effective substitution of one ingredient for another may be achieved. Such substitution can be important in the development of least-cost diets, but the amount of information presently available limits the ability of nutritionists to make such substitutions.

Measurement of digestibility indicates the availability of energy and nutrients in ingredients, providing a rational basis upon which diet can be formulated to meet special standards of available nutrient levels. With this end in mind, this programme is made to evaluate the digestibility of about 40 indigenous fish feed ingredients which ultimately will help to formulate and to prepare nutritional/balanced/quality fish feeds. Gross feedstuff digestibility coefficients will be determined for four species of test fish: common carp, tilapia, rohu and machrocrachium.

2. LITERATURE REVIEW :-

Little work has been done on the digestivility of feedstuffs in our country. In the absence of literature on the digestibility study of feedstuffs on specifc-fishes, the information provided by NRC, 1977 Nutrient Requirements of Warmwater Fishes, and Nutrient Requirements of Warmwater Fishes and Shell Fishes, 1983, National Academy press, Washing to DC, 1983) (Can serve as the basis of determining digestibility of feed ingredients for different species.

The digestibility of nutrients in food is commonly determined by comparing the amount of a insoluble indicator substance such as chromic oxide Cr 2^O3 with the nutrent contet of food and feeds. Digestion co-efficients are then expressed as variations of the formula :

(Windell et al. 1978; E. Austreng 1978; M Jobling 1981 )

The method of analyses of chromic oxide in food and feces developed by Furakawa et al. (1966) has been modified tol limit the use of potentially explosive perchloric acid. Organic material is removed by acid digestion in a solution of hot sulphuric and nitric acids. (Stone, 1982).

The apparent digestibility at different protein may be calculated by the formula :-

(Maynard and Loosli, 1956 )

The method presently used by most fish nutritionist to determine digestibility of feedstuffs is an indirect one using chromic oxide Cr2^O3 as an indigestible marker at 0'5 to 1'0 percent in test diets. The working assumption is that all of the chromic oxide consumed will pass through the digestive tract and appear in the feces. Relative changes in chromic oxide percentage in the feed and feces will represent the percentage of the feed that was digested by the fish. By analysing the feed and feces for their various components ( protein, lipid, energy etc.) the digestibility of each nutrient and the digestible energy of the diet can be determined from the following formula :

Among the studies with warm water fishes that have utilized the above chromic oxide method are those of Hastings, (1966, 1969), Smith and lovell (1971), and Wilson et al (1981).

The determination of digestibility involved measuring the amount of a specific nutrient or feedstuff ingested and substracting that which is present in the feces following digestion. Endogenous material such as secretions from within the intestinal tract, sloughed epithelial cells and other material of metabolic origin may also occur in the feces. Studies on the endogenous or metabolic loses to feces have been performed with rainbow trout by Nose ( 1967 ) and Foltz ( 1978). The later study revealed that the amount of endogenous protien in feces increased from 3'1 percent 6 to 8'4 percent as temperature increased from 7° to 19°C. From a practical standpoint, determining endogenous feces lossesis not considered to be significant.

Nose (1960) collected samples of rectai contents by manually stripping the fish and gently squeezing out the fecal material from the rectum.

Smith ( 1971) confined the fish in metabolic chambers and collected the feces that were voided naturally into the anterior chamber past a latex dam separating the anterior and posterior portion of the trout.

Ogino et al ( 1973) collected the feces from the effluent water and Choubert et al (1979) used a mechanically rotating screen to filter out fecal material.

The “Guelph system (CYAQ-2)” developed by Cho et al (1975, 1982) measures digestibilities by collecting fecal material in a settling column. There are three tanks in each unit, which all drain through a common drainpipe and a single stand pipe placed over an acrylic settling column. The base of the settling column can be surrounded by a cooling Jacket to minimize degradation of the fecal material. It has been observed that larger feces particles are trapped in the settling column within 2 min of being voided by the fish, but some loss of soluble materials may occur.

Windell et al (1978) obtained samples of rectal contents by applying suction to the anus or by disecting the fish. The feces obtained by both of these techniques involved handling the fish and exposing them to considerable stress. Foreed evacuation of their rectum would result in the addition of physiologial fluid and intestinal epithelium to the rectal contents.

Methods of deternining D.E. and M.E. values of feedstuffs for fin fish were reviewed by Smith (1979), collecting fecal material can be accomplished by one of two methods. Wastes can be obtained from the culture tank or aquarium after defecation, or the contents of the lower large intesting can be removed before the material leaves the fish.

Windell et all (1978) found that a considerable amount of hatching occured from trout feces within a short period of time.

Lovell (1977) indicated that nutrient digestibility among feedstuffs varied considerably for different warmwater fishes. This may be particularly true with respect to fishes that live under different environmental conditions.

Jakeshi Nose (1967) collected the fecal sample, combined, mixed throughly and divided into four to six parts to be analyzed for chromium oxide and total nitrogen. Chromium oxide was determined by the wet ashing method with con nitric acid and 60% perchloric acid according to Furalcawa and Tsukahara (1966).

Takeshi Nose (1967) while determining the apparent and true digestibilities of proteins such as milk casein, egg albumin, white fish meal and defatted Soybean meal at different protein on fifty individuals of the rainbow trout fingerlings by the indicator method observed that the lowering of digestibility at lower protein levels is obvious in milk casein, white fish meal and defatted soyabean meal. Egg albunun on the contrary, shows a different pattern from those of the other proteins. The digestibility of egg albumin reveals the highest value, 86% at a 19% protein level and decreases gradually according to the increase of protein level. The some results have been obtained in the previous work (Nose 1962) in which egg albumin attained a maximum value of digestibility, 85% at a 30% egg albumin level but decreased to 71% at 50% level.

Page and Andrens (1973) indicated that decreased protein digestibility occured in channel catfish when the diet had high levels of carbohydrates. Feeding rate can also affect digestibility.

Andrews (1979) found that the DE value of channel catfish test diet improved when the fish were restricted to 90 percent of the control feeding level. The amount of feed consumed did not appear to affect protein digestion but may have influenced that of lipids and carbohydrates.

Andrews et al (1978) found that in channel catfish the apparent digestibility of tallow was 94 percent at 28°c, but that it decreased to 70 percent at 23°c.

Channel catfish digestion was found to be more rapid between 26'6° and 29'4°C than between 21'1°C. and 23'9°C. Shrable et al, (1969). Unpublished data from Japan indicated that protein digestibility in common carp remains relatively constant over the range of temp, that supports growth, but decreases as temp, falls below that range (NRC, 1977).

Starch digestibility appears to be good in warm water fishes. Chiou and Ogino (1975) found that common carp were able to digest about 85 percent of the diet.

Takeuchi (1979) determined that common carp digest over 90 percent of the soybean meal, coconut oil, or Alaska pllock liver oil fed in experimental diets but they digest lard at a level of only 75'6 percent. Digestibility was found to be the same at both 15° and 25°C.

Law et al (1985) worked on the digestibility of carpet grass and napier grass by grass carp and observed that only 20'92 and 16'45% dry matter respectively was digested, while 86'7% of the protein released from the grass was absorbed by the species. However, with pelleted feed, digestibility of dry matter and protein was 82'85 and 90'14% respectively.

Wannigama et al (1985) worked on the protein digestibility by sarotheradon nilotica of 4 diets containing 19,20,25 and 29% crude protein and observed that digestibility decreased with decreaning dietary crude protein levels, except in the case of diet containing 19% crude protein, which did not contin only rice bran. The other diets contained rice bran which was of poor quality.

Sena (1985) carried out digestibility experiments on sarotherodon nilotica fry at different salinities using Cr2°3 as a marker at the rate of 3% and observed that dry matter and protein digestibility varied from day to day, indicating a possible rhythmicity in digestibility. Cho et al (1985) found the apparent digestibility co-efficients of protein ingredients in diets fed to rainbow trout, as blood meal 87% fish meal 92% and soybean meal 96%.

Cho et al (1978) developed a method for measurement of apparent digestibility which may be applied to groups of growing fish of any size and permits the fish to be maintained at normal conditions. This “Guelph system” gave results which were in good agreement with methods based on intestinal dissection and suction with a vaccum for protein and fat. Grude protein and crude fat in most ingredients were well digested. However, the crude protein in blood meal was poorly digested, with that in hydrolyzed feather meal, poultry-by product meal, and tower rapeseed meal being only moderately well digested. The digestibility of pure cerelose and dextrin was high while neither raw nor auto-claved corn starch were digested. On the otherhand, while the starch in raw wheat midlings was not digested, heating this products improved the digestibility of the starch, Starch in soybean meal and corn gluten meal were reasonably well digested.

Silva et al studied the dry matter and apparent protein digestibility co-efficients in sarotherodon nilotica fry of mean weight 21 maintained on four artiticial diets with protein content ranging from 9'6% to 30'4%. They found that the mean, apparent, total protein and energy digestibility of the different groups of fry ranged from 56'7% to 69'1%, 71'7% to 87'1% and 73'6% to 83'9% respectively. The dry matter digestibility in all experimental groups of fry, showed a significant decrease when dietary protein content changed from 9'6% to 30'4%.

Austrng et al (1980) observed the digestibility of different fats and fatty acids in rainbow trout and mink and found that both species responded very similarly to digestibility differences between dietary fats and fatty acids. Both species revelead decreasing digestibility of total lipids and fatty acids with increasing melting point. Soybean oil, cod liver oil and capelin oil were efficiently digested, while hydrogenation of capelin oil resulted in decreased digestibility.

Hanley (1987) studied the co-efficient of digestibility for crude protein and gross energy of several food-stuffs used in Nile tilapia (Oreochromis nilotica 1.) fingerlings sized meals in aquaria fed essentially single ingredient diets containing approximately 80% of the test foodstuffs at a rate of 3% of wet body weight per day and found that the digestibility co-efficients for protein were soybean meal (91%), fish meal (86%), ground corn (83%), wheat middlings (75%), poultry offal meal (74%), brewer grain (63%). In regard to gross energy the digestibility were animal oil. (93%), fish meal (80%), ground -nut corn (76%), poultry offal meal (59%), wheat middlings (58%), soybean meal (56%) and brewers grain (30%). It was apparent that the protein and energy of the animal based foodstuffs were more available to O. nilotica than that of in plant based feedstuffs.

Bergot, F. et al (1983) studied the starch digestibility with two diets containing 70% of a common mixture and 30% corn starch which was either native or gelatinized. The feces were recovered by continuous automatic collector. Restriction and gelatinization promoted a positive effect on starch digestibility. Native starch digestebility rose from 38% at the high intake to 55% at low one, gelatinized starch digestibility which was much higher than that of a native starch, rose from 87 to 90% under the same feeding pattern. Fecal carbohydrates of fish fed gelatinized starch contained a greater proportion of dextrins than those of fish fed native starch.

3. Objectives of proposed work to complement the protein Digestibility of feedstuffs in Nutrition programme B.

1. To develop a technique to measure the digestibility co-efficients in major carps, Tilapia, common carps or freshwater shrimp for lipid and energy components.

   1. Formulation of reference diet and test diet with test feedstuff.

   2. Trial of reference diet and test diet with test feedstuff on selected species under laboratory conditions to collect feces quantitatively.

2. To calculate the apparent lipid and energy digestibility of some (30) locally available plant and animal originated ingredients.

WORKPLAN SUMMARY AND ACTIVITY CHART

WORKPLAN SUMMARY :

It is generally acknowledged that diet formulation should be done on the basis of digestible energy values for the involved spe cies. For the formulation of quality fish feeds digestibility of different feed stuffs on different species should be studied. 30 feedstuffs will be selected from the FARS FNI Project. The group of fish that included in this study will be : Major carp (Rohu), Tilapia (Oreochromis nilotica), Shrimp (Golda), and Common carp c cyprinus carpio. Two diets will be prepared for each digestibility test. One test diet containing the test feedstuffs and another the reference diet. Triplicate lots of fish will be test for feedstuff digestibility.

The feed stuffs will be selected on the basis of

1. Availability and economic aspects.
2. Nutritive value from survey assays and NRC data where available.
3. Acceptability by the fish such as flavour and colour.

The trial on diet will be conducted with fingerlings/ 4“–8” fish.

1. In aquarium under laboratory conditions.
2. In synthetic tanks under pilot plant conditions at FARS.

4. METHODOLOGY :

The digestibility co-efficients will be measured by the indirect method using chromic oxide as a marker. 30 feed ingredients out of 83 analyzed ingredients of Fish Feed Ingredients survey project (FNI, FARS, FRI) will be selected for the study. During the selection of these feed ingredients, nutrition value will be considered and those feed ingredients will be selected which have relatively high nutritional value. Three species of fish will be selected for the trial with reference and test diets containing the test feedstuff at 50% of test diet formula.

A. Feed Ingredients to be used :-

1. For reference diet.
   1. Wheat flour/wheat bran
   2. Rice flour/rice bran
   3. Casein
   4. Egg
   5. Vitamin premix
   6. Chromic oxide (marker)
2. For test Diets : Material to be included as half (50%) of reference diet.

   Kachuripana (small)	------	whole plant
   "              "	------	leaves,
   Kachuripana (large)	------	leaves
   "             "	------	stem
   "             "	------	flowers
   Topa pana	---------	leaves
   Khudi pana	---------	whole
   Kuti pana	---------	whole
   Masur bran
   Mung bran
   Till oil cake
   Mustard oil cake
   Lin seed meal
   Lin seed oil cake
   Soyagrets
   Coconut oil cake
   Cotton seed oil cake
   Molasses
   Panikachu leaves
   Blood meal (cow)
   Blood meal (goat)
   Slaughter house waste		(cow)
   "       "        "		(goat)
   "       "        "		(chicken)

   * If diets are unaccepted, the trial will be repeated with test feedstuff at 25% of reference diet.

   Crab meal
   Prawn shell	----	Galda
   "     "	----	Bagda
   Fish meal	----	A1	(grade)
   "     "	----	A2	"
   "     "	----	B	"
   Fish silage
   Rice bran	----	(Pajam)
   "     "	----	(Irri-20)
   Wheat bran	----	Fine
   "     "	----	Coarse
   Barley
   Silkworm pupae
   Kitchen waste - (Hotel)
   Rice starch

B. a) Formulation of practical diet :-

For the formulation of practical diet emphasis is generally given on the level of protein and energy. One reference diet will be prepared with the above mentioned ingredients A (a) with an optimum level of protein and energy.

The diets will be prepared by reference diet with one fish feed ingredients as 50 : 50 : or 75 : 25 :, depending upon the optimum level of protein and energy. In this way 30 test diets will be prepared. The optimum levels of protein and energy for each group of fish will be in accordance with the recommendations of NRC (1977). The crude protein level in the diet will be balance using the square method described by Hardy (1980).

b) The following species will be selected as test species for trial /study with respect to digestibility.

1. Rohu - Laveo rohita or Mrigal - Cirrhinus mrigala.
2. Common carp - Cyprinus carpio
3. Tilapia - Oreochromis nilotica
   or

4. Freshwater - Macrobrachium rosenbergii

c) A work-sheet for diet formulation will be prepared showing gross nutrition and energy balances.

d) The preparation of practical diets will have the following steps :-

1. Particle size reduction.
2. Premixing,
3. Mixing,
4. Pellating or flaking,

C. a) Experimental design :-

The aim of the diet experiments will be to measure the digestibility of the selected feedstuffs on specific fishes. The trial will be done in two way, First, the trial with reference diet on specific fish and secondly a trial with test diet on specific fish (same). Trials will be set in aquarium/synthetic troughs under laboratory condition. Treatments would be replicated and replicates test would be randomized to obtain a vaild error term co-efficient of variation (CV) will be determined.

Significance of result will be analyzed by ANOVA and simple student T-test for significance between mean values obtained from the chemical analysis and calculated apparent digestibility coefficients.

b) Feeding :

Feeding practices with follow regular feeding procedures, amount and frequency will mainly follow the recommendations made in NRC-1977 with certain modification as necessary. After one week long acclimitization, the feeding of reference diet and test diet will be started. Chromic oxide will be used with feed as a marker to measure the digestibility co-efficients of feedstuff by the indirect method. Feed will be given twice daily at 800 hours and at 1600 hours and will be fed to 5 to 7 days.

c) Trial on laboratory level :

Synthetic troughs and aquarium will be used for feeding trial of reference and test diet on laboratory level. Care will be taken to ensure adequatewater inflows to meet the ultimate requirements of the fish-since some species adapt readily to trough/aquarium and some do if necessary. In the present study, prolonged exposure, more than 12 hours per day, to oxygen levels below 5 ppm will be avoided by using aerator for oxygenation whenever necessary.

In trials of each experimental lot feces will be collected by sacrificing the fish and feces will be collected from the last 25% of the intesting on the 5th or 7th day at 10'00 after 15th application of the feed or at 1800 after 2nd application of the feed. Disecting the fish, feces will be collected on watch glasses by scraping the intestine (last 25%). The fecal samples thus collected will be dried in an over at 105°C overnight (to constant weight)

The dried samples (feces) will be weighed and ground in a mortar. The chromic oxide content of the feces and of the diets will be determined by using the wet-acid oxidation method. Apparent digestibility co-efficients will be calculated by using the formula of Maynard and loosli (1956), employing chromic oxide as the dietary marker.

Apparent digestibility of the feedstuff will be calculated based on content of feedstuff used in the test diet.

ie DC_TF = (DC_TD - 0.5 DC_RD)₂ when a 50/50 mix is used following gross feedstuff digestibility feedstuff used and feeal samples will be analyzed for crude protein, total lipid, ash and fiber to determine apparent digestibility of protein fat and energy components.

* Will be avialable in the fish Nutrition laboratory, FARS, FRI

Bibliography

AOAC. 1975. Official Methods of Analysis of the Association of Official Analytical Chemists. 12th edition. Association of Official Analytical Chemists. Washington DC. 1094 P.

Andrews, J.W., M.W. Murray and J.M. Davis, 1978. The influence of dietary fat levels and environment al temperature on the digestible energy and absorbability of animal fat in catfish diets. J. Nutr. 108 : 759–752.

Andrews, J.W. 1979. Some effects of feeding rate on growth, feed conversion and nutrient absorption of channel catfish. Aquaculture. 16: 243–246.

Austreng, E., 1978. Digestibility determination in fish using chromic oxide marking and analysis of contents from different segments of the gastrointestinal tract. Aquaculture, 13: 265–272.

Chiou, T.Y. and C. Ogino, 1975. Digestibility of starch in carp. Bull. Jpn,Soc,Sci,Fish. 41: 465–466.

Cho. C.Y., Slinger, S.J. and Bayby, H.S, 1982. Bioenergetics of salmonid fishes, energy intake, expenditure and productivity, Comp, Biochem. physiol., 73 B : 25–41.

Choubert, G., Jr., Dela Noiie, J. and Luquet, P. 1979. Continuous quantitative automatic collector for fish feces. Prog. Fish cult. 41 : 64–67.

De Silva, S.S. and Perera. M.K. 1983. Digestibility of an aquatic macrophyte by the cichild Etroplees suratensis (Bloch) with observations on the relative merits of three indigenous components as markers and daily changes in protein digestibility. J. Fish Biol., 23: 675–684.

Foltz, J.W., 1978. The effects of meal size and temperature on gastro-intestinal motility and absorption in rainbow trout (Salmo gairdneri) and tilapia (Sarotherodon mossambicus) Ph.D. dissertation. University of colorado, Boulder., 62 P.

Furukawa, A. and Tsukahara, H., 1966. On the acid digestion method for the determination of chromic oxide as an index substance in the study of digestibility in fish feed. Bull. Jpn. Soc. Sci., Fish., 32: 502–506.

Hastings, W.H., 1969. Nutritional scure. In: O.W. Neuhaus and J.E. Halver (Editors). Fish in Research, Academic press, New York, PP. 263–292.

Jobling, M. 1981. Dietary digestibility and the influence of food components on gastric evacuation in plaice, pleuonectes platessa. J. Fish. Biol. 19: 29–36.

Maymard, L. A. and Loosli, J.K., 1956. Animal Nutrition MCG raw Hill, New York, 4th ed., 484 PP.

Nose, T. 1960. On the digestion of food protein by gold fish (carassium auratus L.) and rainbow trout (Salmo iridens G.) Bull. Freshwater. Fish. Res. Lab. 10 : 12–22.

Nose, T, 1967. Recent Advances in the study of fish Digestion. PP. 83–94 in symposivm on feeding in trout and salmon culture, J.L. Gaudet, ed EIFAC Technical paper No. 3. Rome : FAO.

NRC, 1977. Nutrient Requirements of warmwater fishes. Nutional Research Council, National Academy of Sciences, Academy press, Washington DC.

NRC, 1983. Nutrient Requirements of warmwater Fishes and shell-fishes. National Research Council National Academy of Sciences, Academy press, Washington DC.

Page. J.W., and J.W. Andrews, 1973. Interaction of dietary levels of protein and energy on channel catfish (Ictaturas Punctatus), J. Nutr. 102: 1339–1346.

Shrable, J.B., O.W. Tiemeir and C.W. Deyoe, 1969. Effects of temperature on rate of digestion by channel catfish. Prog. Fish cult. 31: 131–138.

Smith, B.W. and Lovell, R.T., 1971. Digestibility of Nutrients in semi-purified rations by channel catfish in stainless steel troughs. Proc. 25th Annu. coat§. Southeast Assoc. Game Fish comm., PP. 452–459.

Smith, R.R., 1971. A method for measuring digestibility and metaboli metabolizable energy of fish feeds. Prog. Fish cult., 33 : 132–134.

Smith, R.R., 1979. Methods for determination of digestibility and metabolizable energy of feedstuffs for finish. In J.R. Halver and K.Tiews (Editors), Proc. World Symposium on Finfish Nutrition and Fish feed Technology, Vol. II Heenemann, Berlin, PP. 453–459.

Takeuchi, M. 1979. Digestibility of dietary lipids in carp. Bull. Tokai Regional fish Res. Lab. 99: 55–63.

Wilson, R.P; E.H. Robinson and W.E. Poe. 1981. Apparent and true digestibility of amino acids from common feed ingredients for channel catfish. J. Nutr. 111: 923–926.

Windel, J.T.: J.W. Foltz and J.A. Sarokon, 1978. Methods of fecal collection and nutrient leaching in digestibility studies. Prog. Fish cult. 42: 51–55.

ANNEX D

NUTRITION AND DIET DEVELOPMENT PLANS 1989–1991

ANNEX E

C-6: SUPPLEMENTARY FEEDS FOR POLYCULTURE

OBJECTIVES:

1. Determine limiting nutrient in polyculture;
2. Determine cost-effective supplementary feed;
3. Measure increased production in polyculture of fishes with supplementary feeds.

DESIGN:

FEED 1% BODY WEIGHT PER DAY

MEASUREMENTS:

1. Monthly biomass in ponds to adjust feed;
2. Monthly gain;
3. End weights, population, feed conversion;
4. Mean production cost, w/o treatment;
5. Mean production cost, w/o fertilizer;
6. PER for diet treatments;
7. NPO for diet treatments;
8. ANOVA and INTERACTION statistics;
9. Cost/kg for gains with supplementary feeds.

ANNEX F

SPECIFICATIONS FOR FREEZER-COOLER EQUIPMENT

Bidder shall furnish one industrial type 2 stage ammonia type freezer compressor with 440V 3phase V belt drive motor, complete with compression valves and guages plus one ammonia refrigerate storage tank, expansion freezer coils with blast freezer fan with automatic and mannual override controls, together with floor mounted condensor unit to operate one 30 cubic meter insulated coldroom to operate at -20 to -5 degree Celcius. Coldroom will be constructed and provided by purchaser. In addition bidder shall supply one industrial type single or double stage ammonia type compressor with 440V 3phase V belt drive motor complete with compression valves and guages, common or independent liquid ammonia storage tank, expansion coils with automatic and mannual override controls, floor mounted condensor and expansion coils with fan to operate one 20 cubic meter cool room at +2 to +5 degrees Celcius. Cool room will be constructed and provided by purchaser. Bidder shall provide detailed description, design and instructions for installation of equipment for independent or coupled operation. In addition bidder shall provide two 1m × 2m industrial type insulated freezer room doors complete with heavy duty steel frames and all hardware plus international SAFETY emergency escape plunger release latch; doors to be installed by purchaser at site. Equipment furnished shall be equal or better than the following: (1) Howe 3×3 ammonia compressor with base 5hp 440V 3P motor; (1) Howe 5×5 compressor with vbase and 15hp 440V 3-P motor; (1) Jpwe vertical condensor for gravity feed; (1) Howe 12"×10' receiver; (1) Howe UC 182 unit cooler with surge drum and controls equiped with water defrost; (1) Howe UC 183 blast freezer with surge drum and water defrost controls; together with all necessary valves, piping, tubing, belts, and spares necessary for one year of continuous operation. Control panel shall be equal to or better than Howe control panel with motor starters and defrost controls.

Bidder shall provide all above specified materials and supplies plus descriptive details for installation of above equipment by qualified local personnel within the areas outlined in engineers sketch of facilities.

ANNEX G

MINIMUM EXPECTED FEED MILL EQUIPMENT

* ( ) = specification item number Annex H.

ANNEX H

SPECIFICATIONS FOR EXPERIMENTAL FEEDMILL EQUIPMENT

(Supplier will include descriptive literature with each submission of bid).

(All prices are F.O.B. Seattle, USA, price estimates as per 01-06-89).

1. Counterbalance scale. Counterspring dial scale with chart. 50 kg capacity X 100 g and 10 g unit scale, stainless steel construction, mounted on wheeled stand. Similar or equal to Hobart model HOV 15. Delivery 60 days. $ 1,500.

2. Twin-shell blender. Twin-shell liquid-solid blender with ⅓ - ¼ horsepower 220 volt, single phase shell drive; and 1 horsepower, 220 volt, single phase motor activator bar drive; with one plastic shell and one stainless steel shell, 5 kg capacities. Similar or equal to Paterson-Kelly Company model LB-8T. Delivery 60 days. $ 6,000.

3. Meat grinder/meat chopper. Grinder/chopper with 2 – 3 horsepower, with 220 volt, single phase or 440 volt 3 phase 50 cycle electric motor with thermal overload protection and stainless steel feed pan, together with 2 mm, 4 mm, 6 mm steel plate dies, 4 cutter knives, machine mounted on casters. Similar or equal to Hobart Meat Chopper model 4632. Delivery 60 days. $3,000.

4. Extruder. Semi-moist pellet extruder, 1 horsepower, 220 volt, single phase, 50 cycle electric motor with overload protection, stainless steel loading chute, 20 kg capacity with mixing arm and spiral screw auger and safety switch cover grill. Machine to be supplied with 2 mm, 3 mm, 4 mm, and 6 mm plates and variable speed external pellet clipper. Similar or equal to OMR Fidenza model Parmigiana spaghetti/noodle machine. Delivery 90 days. $ 7,500.

5. Micropulverizer. Micropulverizer with variable speed drive, visable ammeter and thermal overload protection switch. One horsepower, 220 volt, single phase, 50 cycle electric motor with ¼ horsepower 220 volt, single phase, 50 cycle variable speed auger drive, complete with 1 mm, 2 mm, 3 mm, and 4 mm round and 45 degrees angle cutting screen, and one extra set of mill hammers, stainless steel or brass construction, complete with one liter sample collecting or linen bag discharge chute collecting devices. Similar or equal to Sich Industrial Company micropulverizer type CF. Delivery 60 days. $9,000.

6. Vibrating screen sifter. Circular vibrating sifter machine, 1 horsepower, 220 volt, single phase, 50 cycle, with set of 0.05 mm, 0.1 mm, 0.15 mm, 0.2 mm, 0.25 mm, 3 mm and 4 mm screens, 30 cm in diameter X 10 –15 cm height with individual discharge chutes and set of stainless steel adjustable bands for mounting 2 to 2–6 screens equiped with 5 cm × 1 cm circular screen brushes and screen feed chute. Similar or equal to Baldor Sweco separator model LS 185. Delivery 60 days. $6,000.

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8. Cabinet dryer. 1 m × 1 m × 2 m cabinet with 4 shelves and forced air drying, equiped with 5 kilowatt heater, 220 volt, single phase, 50 cycle electric motor, temperature range ambient to 150 C complete with temperature control thermostat and horizontal air flow; stainless steel construction. Similar or equal to Meatpackers and Butchers Supply bortron mini-dryer, 200 pound capacity. Delivery 60 – 90 days. $ 10,000.

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10. Dough mixer. 220 volt, single phase, 50 cycle variable speed mixer complete with one 20 liter stainless steel bowl, one 30 liter stainless steel bowl, splah covers for each, dough arm, flat beater, and wire loop whip. Similar or equal to Hobart sales model A200 blender. Delivery 60 days. $ 2,500.

11. Hammer mill. 2 horsepower, 220 volt, single phase, 50 cycle, v-built drive mill equiped with fan, 2 screens, loading chute, collecter pipe, discharge tube, and permanent magnet built into hopper plate. Mill to include 2 sets of 24 hammers and 2 sets of 4 hammers with pivot rods and pins plus set of screens at 0.25 mm, 0.5 mm, 1.0 mm, 1.5 mm, 2 mm, and 3 mm together with 6 backing screens for above. Similar or equal to Meadows Mill Company model Mighty Marvel #5 with accessories. Delivery 30 days. $ 3,000.

12. Ribbon mixer. Horizontal, double spiral ribbon mixer, stainless steel construction, 100 kg flour capacity, front dump, reversable drive, with safety switch on cover. Sanitary meat-type construction with standard packing glan seals and gasketted quick-open cover clamps each with limit switch. Mixer tub ends to be formed with full 2 cm radius, and all interior well continuous and ground smooth, 3 horsepower, v-built drive, 220 volt, single phase, reversable motor or 440 volt, 3-phase, 50 cycle reversable motor. Ribbon mixer side clearance to be not more than 2 mm, and machine to be mounted on casters with roller locks. Similar or equal to Hous Company sani-mix ribbon mixer model S-204. Delivery 90 days. $16,000.

13. Compressed pellet mill. Laboratory size pellet mill for production of compressed pellets, variable speed screw drive, mechanically agitated feed hopper, caster table mounted with totally enclosed 2 horsepower, 440 volt, 3-phase, 50 cycle main drive motor equiped with sheaves and v-built drive for 4 die speed rotations; 1/4 horsepower, 220 volt, single phase, 50 cycle feeder/mixer drive motor together with visable ammeter, thermally protected overload starters, pushbutton controls, and manually operated major disconnect. All contact parts to be 302 – 304 stainless steel, except parts hardened to 410 stainless steel or hard chrome plated. Mill complete with steam injection fittings, and to be provided with 2 mm, 3 mm, 4 mm, 6 mm, and 8 mm dies of stainless steel or hard alloy steel. Similar or equal to California Pellet Mill (CPM) model CL type 2. Delivery 90 days. $ 30,000.

14. Double drum dryer. Laboratory model 150 psi steam capacity, equiped with two 20 cm counter-rotating stainless steel or hard chrome finish drums and variable speed drum drive, 1/2 horsepower, 220 volt, single phase, 50 cycle motor with drive range of 1 to 10 rpm, and 2 stainless steel doctor blades together with collecting chutes and fiberglass or hard plastic adjustable drum surface guides. Drum to be adjustable for gap clearance of 0 to 6 mm. Machine to be provided with visable steam pressure gauge, and variable steam pressure control valve. Similar or equal to Meldon Associates buflovak double drum dryer, laboratory model, atmospheric type. Delivery 120 days. $ 11,000.

15. Plastic bag vacuum sealer. Table-top model vacuum sealer with 1 horsepower, 220 volt, single phase, 50 cycle or 440 volt, 3-phase, 50 cycle motor, internal vacuum pump, and adjustable temperature heater bar, sealing chamber not less than 10 cm × 25 cm × 25 cm. Similar or equal to Meatpackers and Butchers Company piccolo vacu-fresh table-top sealer # 42009. Delivery 60 days. $4,000.

16. Sample splitter. Sample splitter for particles and ground material, stainless steel construction. Similar or equal to Tyler Incorporated model SS50 Sample Splitter #1. Delivery 60 days. $ 500.

17. Steam generator. Electric, 5 horsepower, 440 volt, 3-phase, 50 cycle 150 pounds per sq. inch steam capacity generator, complete with visable steam pressure and water level gauges, automatic water make-up, and thermal protector switch, excess pressure steam escape valve, and low water automatic cutoff. Similar or equal to Chromalox Sussman steam generator model CMB 4LC1. Delivery 60 days. $5,000.

18. Steam jacketed pot. Steam jacketed pot of stainless steel construction, with 10 cm front dump and lid, with paddle stirrer; double jacketed, floor mounted with visable external steam pressure gauge, steam pressure control valve, with steam capacity of 0 – 80 psi, equiped with condensate return line, and insulated lid handles and dump valve handle. Pot to be mounted on stand with 30 cm floor clearance for collecting vessels. Lid stirrer to be powered ½ horsepower, 220 volt, single phase variable speed electric motor. Internal surface shall be smooth without seams except for dump pipe and mounting lid, with circular sidewalls and spherical bottom for complete and easy cleaning. Internal capacity 100 liter, external jacket without insulation. Similar or equal to Dohrmann Hotel Supply steam jacketed pot, front dump, 30 gallon capacity, floor mounted, uninsulated. Delivery 60 – 90 days. $ 6,500.

19. Pellet crumbler. 220 volt, single phase or 440 volt, 3-phase, 50 cycle motor operating one live-roll and one adjustable idle corrogated roll, 10 cm diameter × 25 cm hardened steel rollbars, and 2 sloping shaker screens of 2 mm, 4 mm, and 6 mm openings with individual discharge chutes and screen over return chute. Laboratory model feed pellet crumbler. Similar or equal to Meatpackers and Butchers Supply pellet crumbler, laboratory 12" model. Delivery 60 days. $ 3,000.

COMMENT: These are current prices as of June 1986, F.O.B. Seattle, USA. The total price of $ 83,250.00 does not include packing for shipment or transportation to Bangladesh. Individual orders to these companies may require 25 % to 50 % pre-payment with order. A recognized company like Halver Corporation or F.D.T. Inc. may be able to order without pre-payment, upon award of contract.

ANNEX I

MATERIALS FLOW DIAGRAM EXTRUDED FEEDS

MATERIALS FLOW DIAGRAM SEMI-MOIST FEEDS

MATERIALS FLOW DIAGRAM PELLET FEEDS

SCHEMATIC MULTIPURPOSE FEED MILL

ANNEX J

MATERIALS FLOW FOR FISH PROTEIN HYDROLYSATE

ANNEX K

MATERIALS FLOW FOR FISH SILAGE