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D.P. Bureau and C.Y. Cho Fish Nutrition Research Laboratory Dept. of Animal and Poultry Science University of Guelph, Guelph, Ontario, Canada, N1G 2W1
Collection of fecal material Measurement of digestibility is an important step in the evaluation of a feed or an ingredient. Digestibility is, in general, a good measure of the nutritive value of a feedstuff and provides a rational basis for the formulation of diets. There is evidence that the collection of all the feces produced by a group of fish is not a feasible task. Digestibility measurements in fish must, therefore, rely on the collection of a representative sample free of uneaten feed particles and the use of a digestion indicator (indirect method). Several techniques have been used to collect fecal material from fish. Several of these techniques, still in use for practical reasons or by preference, will generally not produce reliable (realistic) estimates of digestibility. The most widely used of these techniques is the collection of feces from the lower part of the intestine by stripping, suction or intestinal dissection. This technique generally leads to an underestimation of the digestibility of nutrients (protein in particular) because of contamination of feces with endogenous material that would otherwise be reabsorbed before the feces are excreted (Cho et al. , 1982; Hajen et al., 1993). Other techniques, such as the periodical collection of feces by siphoning from the bottom of a tank, are also likely to yield inaccurate estimates of digestibility since the break up of feces by fish movement may lead to leaching of nutrients and, therefore, overestimation of digestibility of nutrients. Finally, the technique of Smith (1971), where the feces voided into the water are collected from fish immobilised in small metabolic chambers, yields questionable estimates of digestibility, since the fish need to be force-fed, they often regurgitate, and show in negative protein and energy balances. There are three methods/systems that are more likely to produce meaningful estimates of digestibility of nutrients if used correctly: the systems of Ogino et al. (1973) in which the feces are collected by passing the effluent water from the fish tanks through a filtration column (TUF column), Cho et al. (1975) in which a settling column is used to separate the feces from the effluent water (Guelph system) and Choubert et al. (1979) in which a mechanically rotating-screen is used to filter out fecal material (St-Pee system). These three systems have been adopted in several laboratories around the world. These two systems apparently yield very similar apparent digestibility coefficients (ADC) of protein and lipid for several ingredients. In a study comparing the TUF column and the Guelph system, very similar ADC of dry matter, protein, lipid and energy were obtained with both methods for two reference diets (Satoh et al., 1992). With the Guelph system, the fecal material is collected in a settling column as shown in Figure 1. There are three tanks in each unit, which all drain through a common drainpipe, and a single standpipe placed over an acrylic settling column (10 cm diameter x 40 cm high). The base of
Bureau and Cho. 1999. Measuring Digestibility in Fish. UG/OMNR Fish Nutrition Research Laboratory 1 Technical Document, University of Guelph, Ontario, Canada.
nitrogen. UG/OMNR Fish Nutrition Research Laboratory 2 Technical Document. The feces are then freeze-dried and ground for determination of the dry matter. Ontario. Bureau and Cho. One criticism of this method is that soluble material could be lost from the feces due to leaching. no significant leaching losses were observed when feces stood in the Guelph settling collector or TUF column for 15h in freshwater (Satoh et al. gross energy and digestion indicator contents. it has been observed that larger feces particles are trapped in the settling column within 2 min of being voided by the fish. University of Guelph. however. and carcass analysis are made. which must be avoided (Windell. The feces are centrifuged at 5. allowing repeated sampling over 6-9 days. The fish are accustomed to both the tanks and the dietary regime for at least 3 days before collection of feces begins. to avoid wastage. Canada. Moreover. one unit being devoted to the reference diet. The velocity of the water flow is adjusted to minimise settling of the feces in the drainpipe and maximise recovery of the feces in the settling column. 1999. Each tank is loaded with 2-4 kg of fish. the settled feces and surrounding water are gently withdrawn from the base of the settling column into a large centrifuge bottle by gravity. The close agreement between ADC of dry matter. growth rate.. lipid. One-third of the water in the tanks is drained out to ensure that the cleaning procedure is complete. ash. The fish are under the rearing conditions normally applied to other nutritional experiments. 1992). Immediately after collection of the feces.000 x g for 10-20 min and the supernatant discarded. the fish are fed again as normal. The tanks each measure 55 cm x 40 cm and have a sloped bottom. indicate that leaching is not an important source of error. A series of eight such units allows the determination of ADC for up to seven ingredients at any one time. The fish maintain normal feed intake and growth rate and thus allow for the measurement of ADC values which are realistic and applicable to a normal growing conditions.the settling column can be surrounded by a cooling jacket to minimise degradation of the fecal material. crude protein. The main cause of leaching is the break up of feces particles that results from physical handling to remove from water (handling loss). These feces are free of uneaten feed particles and are considered to be a representative sample of the feces produced throughout the 24-hour period. . The Guelph system allows repeated determinations. 1978). Under normal operation. The fish are fed three meals daily between 0900 and 1600 hours. the diets being offered only as long as the fish are actively feeding. the drainpipe and the settling column are brushed out to remove uneaten feed particles and feces from the system. and lipids obtained with TUF and St-Pée systems with the results obtained using the Guelph system. One hour after the last meal. Measuring Digestibility in Fish. At 0830 hours the following day. and evaluation of different diets can be carried out at the same time as observations of apparent digestibility.
Estimating Digestibility of Individual Feed Ingredients Very few potentially useful feed ingredients can be fed voluntarily as the sole component of a diet to fish. UG/OMNR Fish Nutrition Research Laboratory 3 Technical Document. determination of the digestibility of an ingredient requires comparison of the ADC of a reference and a test diet. (1985). . the test diet being a mixture of the reference diet and the test ingredient (Table 1).Figure 1. Thus. 1999. Canada. Measuring Digestibility in Fish. Bureau and Cho. University of Guelph. Ontario. Guelph settling column system (CYAQ-2) Source: Cho et al. Inclusion of a digestion indicator in the diets allows the digestibility coefficients of the energy and nutrients in the diets to be calculated from measurement of the ratios of nutrient to indicator in the diet and feces. It is always necessary to combine a mixture of feed ingredients to formulate a nutritionally balanced diet that will support good feed intake and normal physiological function.
Canada. 3% fiber) Wheat middlings (17% CP. University of Guelph. UG/OMNR Fish Nutrition Research Laboratory 4 Technical Document. Measuring Digestibility in Fish. 1985) _________________________________________________________________ Ingredient % _________________________________________________________________ Fish meal. 1999. 8% fiber) Vitamin premix (VIT-8404) Mineral premix (MIN-8204) Digestion indicator 30 17 13 27 1 1 1 Fish oil. 3% fiber) Corn gluten meal (60% CP. . herring or capelin 10 _________________________________________________________________ Experimental Diets _________________________________________________________________ Reference diet Test diet _________________________________________________________________ (%) Reference diet 100 70 Test ingredient 0 30 _________________________________________________________________ Bureau and Cho. <13% ash) Soybean meal (48% CP.Table 1. Reference diet for salmonids (Cho et al. Ontario. herring or capelin (>68% CP.
0. . 0.ADCR) (2) where ADCI = Apparent digestibility coefficient of test ingredient ADCT = Apparent digestibility coefficient of test diet ADCR = Apparent digestibility coefficient of the reference diet DR DI DT s 1-s = % nutrient (or kJ/g gross energy) of the reference diet mash = % nutrient (or kJ/g gross energy) of the test ingredient = % nutrient (or kJ/g gross energy) of the test diet = Proportion of test ingredient in test diet mash (i.e. (1998) Bureau and Cho. UG/OMNR Fish Nutrition Research Laboratory 5 Technical Document. University of Guelph. 1982): ADC= 1-(F/D x Di/Fi) where D F Di Fi = % nutrient (or kJ/g gross energy) of diet = % nutrient (or kJ/g gross energy) of feces = % digestion indicator (AIA) of diet = % digestion indicator (AIA) of feces (1) ADC of the test ingredients (ADCI) is then calculated based on the digestibility of the reference diet and the test diets as follows: ADCI = ADCT + ((1-s) DR / s DI) (ADCT . Measuring Digestibility in Fish. Canada.7) Sugiura et al.3) = Proportion of reference diet mash in test diet mash (i. 1999.e.. Ontario.The apparent digestibility coefficients (ADC) for the nutrients and energy of the test and reference diets can be calculated as follows (Cho et al.
There is interest in quantifying the ENL in order to be able to calculate the “true” digestibility of protein and amino acids in feeds and ingredients. Under these conditions. Measuring Digestibility in Fish. together with the residues of the microflora which inhabit the digestive tract (Nyachoti et al. Conversely. 1997.Apparent vs.. it is reasonable to doubt of the accuracy of “true” protein digestibility coefficients calculated using estimates of ENL obtained from fish fed protein-free diets. These residues are the remains of mucosal cells. the contribution of ENL to total fecal nitrogen is probably small. Accurate estimation of ENL may require the use of more sophisticated techniques (see Nyachoti et al. For these reasons. University of Guelph. mucoproteins and other secretions released into the digestive tract by the animal. Ontario. antinutritional factors) can enhance ENL (Nyachoti et al. Moreover. and could produce unreliable estimates of apparent digestibility. there is evidence that the amount of ENL produced by animals receiving a protein-free diet differs significantly from that of animal fed diets containing protein. Salmonids will not accept (or will eat very little of) a protein-free diet making it very difficult to calculate meaningful estimates of ENL. In fish maintaining a high feed intake. the difference between the "true" and apparent digestibility of protein is negligible (Hardy. which in turn allow the measurement of apparent digestibility values which are reliable and repeatable. digestive enzymes. true digestibility Feces are composed of the undigested food components and the unreabsorbed residues of body origin. several other dietary constituents (fiber. UG/OMNR Fish Nutrition Research Laboratory 6 Technical Document. . 1997). 1997). it is preferable to discard the fecal samples collected since these samples may contain a high proportion of ENL. 1997 for review). (1982) generally allows the fish to maintain high feed intake and good growth rate. Canada. The use of nutritionally balanced reference and test diets (Table 1) as proposed by Cho et al. Bureau and Cho. if poor feed intake or poor growth are observed in a digestibility trial.. Figure 2).. 1999. In addition. This type of work remains to be carried out with fish. The fecal nitrogen not directly originating from ingested food but from the animal itself are referred to as endogenous nitrogen gut losses (ENL).
The apparent digestibility of good quality protein by fish is very high. Canada. The physical state of some ingredients may affect the digestibility of protein. but dehulled canola meal (canola meat) has a significantly higher ADC for protein (Hilton and Slinger. The same phenomenon can occur with fish meal (Anderson et al. Fish have different digestive capabilities compared to terrestrial animals. 1990. Table 2. 1986). particularly cereal grains and their by-products which contain high levels of starch and fiber. Apparent vs "true" (N-corrected) digestibility (theoretical relationship only) Estimates of apparent digestibility for salmonids Table 2 presents the apparent digestibility coefficients for commonly used ingredients in salmonid feeds as measured by Cho et al. Measuring Digestibility in Fish.. The type of drying technique used during processing is a very important factors. Apparent digestibility coefficients of ingredients measured with rainbow trout (Cho et al. and many feedstuffs. UG/OMNR Fish Nutrition Research Laboratory 7 Technical Document. . University of Guelph. are very poorly digested by carnivorous fish. For example. The protein digestibility of flame-dried blood meal is very low whereas the digestibility of spray-dried blood meal is very high. canola meal has a rather low ADC of protein. Ontario. Apparent digestibility coefficients (%) Ingredients Dry Matter Crude Protein Lipid Energy Bureau and Cho. 1982. 1993).Figure 2. Bureau et al. Cho and Kaushik. However.. several factors can affect the digestibility of protein. (1982). 1999. 199x). A good demonstration of this is seen in blood meal (Table 2).
1982. Hamburg. Cho. Soybean meal. Cowey.. Apparent digestibility measurement in feedstuffs for rainbow trout.. G. dehydrated Fish protein concentrate Soy protein concentrate 39 87 91 55 76 23 23 80 46 77 85 70 76 35 78 74 35 97 90 77 87 85 96 16 91 95 92 96 85 77 92 85 89 77 96 96 92 96 95 97 71 71 97 94 - 43 86 92 50 77 39 29 83 51 77 91 80 82 45 85 75 46 94 94 84 References Anderson. A. dehulled Wheat middlings Whey.Y. University of Guelph. Physiol. Methodological approaches to research and development.J. C. II. 1990.Alfalfa meal Blood meal ring-dried spray-dried flame-dried Brewer’s dried yeast Corn yellow Corn gluten feed Corn gluten meal Corn distiller dried soluble Feather meal Fish meal. Apparent digestibility of rendered animal protein ingredients for rainbow trout (Oncorhynchus mykiss). Cho. Aspects of Food Production. Aquaculture. Anderson and M. and T.B. cook. 125: 305-325. J. S. Bureau. Finfish Nutrition in Asia. Lall. C. Slinger and H.P. Slinger. expenditure and productivity. 1979. Bureau and Cho. (ed. Nutritional energetics in fish: energy and protein utilization in rainbow trout (Salmo gairdneri).H. Vol. S. Germany. Ontario. Kaushik. p. . Harris. Measuring Digestibility in Fish. C. 239-247. Bayley. and S. C. Watanabe. Ottawa.J.S. S. In: Bourne.J. 73B.. full-fat. on Finfish Nutrition and Fishfeed Technology. and S. Biochem.M.A.J. 1993.M. Proc. Cho. McNiven. International Development Research Centre. Cho C. 1999.P. UG/OMNR Fish Nutrition Research Laboratory 8 Technical Document.. 199x. Aquaculture (in press) Cho.Y. and C. herring Meat and bone meal Poultry by-products meal Rapeseed meal Soybean.S. World Symp.).Y.Y. Evaluation of protein quality in fish meals by chemical and biological assays. 154p. 1985.Y. Canada. D. Comp. 25-41. Bioenergetics of salmonid fishes: Energy intake. D.
112: 321-332. Fish. Methods of fecal collection and nutrient leaching in digestibility studies. 1123-1127.K. J. Sci.T.W. Higgs. 33. Apparent protein digestibility and mineral availabilities in various feed ingredients for salmonid feeds. Fish-Cult. Sugiura. Fish-Cult.. Fish-Cult. C. 43: 1149-1155. Bureau and Cho. S. Sarokon.61. 1993b. Smith. Beames. Diet. . J. G. Hajen. Digestibility and utilization of canola meal in practical type diets for rainbow trout (Salmo gairdnerii). 1971.A. Measuring Digestibility in Fish. Satoh. and S. 1997. J. J.Consumption and Energy Values. McBride and H. Karger. and J.. Dong.E. de la and Luquet. 1997. J. Nutr.H. de Lange. 1992. R.M. Can. R. 51-55. Prog. Prog. 149163.... Nyachoti. S. 64-67. Windell. Canada. W.M. Slinger. Aquaculture. University of Guelph. Basel.S. C. Prog.. vol. 1. 132-134. Digestibility of various feedstuffs by post-juvenile chinook salmon (Oncorhynchus tshawytscha) in sea water. Validation of technique.W. Hilton. Ontario. 1978. pp. and T.132-172. D.Jr. A method for measuring digestibility and metabolizable energy of feeds. World Rev. Nippon Suisan Gakkaishi 58. Understanding and using digestibility coefficients in fish nutrition. Significance of endogenous gut nitrogen losses in the nutrition of growing pigs: A review. Aquaculture Magazine (May/June 1997): 84-88.W. 1998. Schulze. Dosanjh. Foltz.M. Watanabe.M. Choubert. Cho.W. R. UG/OMNR Fish Nutrition Research Laboratory 9 Technical Document. F. and R. Aquat. Hardy. and B. C.R. P. J. Anim. Hardy. 41. 40.W. Noue. B. Aquaculture 159. 1999..F. 1986.J. 177202. Can. (1979) Continuous quantitative automatic collector for fish feces.Y. C. Rathbone.A.. Effect of fecal retrieval timing on digestibility of nutrients in rainbow trout diet with the Guelph and TUF Feces Collection Systems. Sci.
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