Aquaculture 207 (2002) 79 95 www.elsevier.

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Apparent digestibility coefficients of various feed ingredients for juvenile and grower rockfish (Sebastes schlegeli)
Sang-Min Lee*
Faculty of Marine Bioscience and Technology, Kangnung National University, Kangnung 210-702, South Korea Received 16 April 2001; received in revised form 20 July 2001; accepted 20 July 2001

Abstract Apparent digestibility coefficients of dry matter, crude protein, energy, and amino acids in white fish meal, anchovy meal, meat meal, feather meal, blood meal, soybean meal, corn gluten meal, cottonseed meal, wheat flour, and brewers yeast were determined for juvenile and grower rockfish. Apparent digestibility coefficients were determined using a reference diet with chromic oxide indicator and test diets that contained 70% reference diet, by weight, and 30% of the feed ingredient being evaluated. The juvenile and grower fish averaging 30 and 300 g were held in 200 l tanks at a density of 50 and 8 fish per tank, respectively. Feces were collected from three replicate groups of fish using a fecal collection column attached to fish rearing tank. Apparent dry matter digestibility of ingredients decreased as nitrogen-free extract or fiber contents of ingredients increased, ranging from 73 95% for animal products and corn gluten meal to 32 54% for soybean meal, cottonseed meal, wheat flour and brewers yeast. Apparent energy digestibility of ingredients followed similar trends to differences in dry matter digestibility. Apparent protein digestibilities of animal and plant products ranged 63 95% and 73 92%, respectively. White fish meal, anchovy meal, meat meal, and corn gluten meal showed the higher protein digestibility among ingredients tested ( P < 0.01), regardless of fish size. Apparent digestibilities of protein and energy were not affected by fish size ( P > 0.05), except for feather meal. These data provide more precise information concerning nutrient and energy utilization of rockfish and will allow ingredient substitutions in practical feed based on levels of available nutrients. D 2002 Elsevier Science B.V. All rights reserved.
Keywords: Rockfish; Digestibility; Feed ingredients; Sebastes schlegeli

Tel.: +82-33-640-2414; fax: +82-33-640-2410. E-mail address: smlee@knusun.kangnung.ac.kr (S.-M. Lee).

0044-8486/02/$ - see front matter D 2002 Elsevier Science B.V. All rights reserved. PII: S 0 0 4 4 - 8 4 8 6 ( 0 1 ) 0 0 7 5 1 - 7

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1. Introduction Rockfish is an important mariculture species in Korea (Lee and Lee, 1994a). Aquaculture production of this species has rapidly increased during the last decade in Korea because it has several desirable characteristics for culture such as good tolerance to low water temperature and ease of seedling production due to the viviparous reproduction style. Several studies on rockfish nutrition have revealed its requirements of protein, essential fatty acids, vitamin E and phosphorous for normal growth (Lee and Lee, 1994a; Lee et al., 1994, 1998; Bai and Lee, 1998). Most marine carnivorous fish require high dietary protein levels compared to omnivorous or herbivorous fish, and fish meal is well recognized as the best dietary protein source for fish (NRC, 1993) including rockfish (Lee et al., 1996). Since fish meal is becoming increasingly expensive and a stable supply of high quality fish meal is becoming difficult, replacement of fish meal protein with alternate protein sources could be of considerable economic advantage. Information on digestibility coefficients of feed ingredients is very useful not only to enable formulation of diets that maximize the growth of fish by providing appropriate amounts of available nutrients, but also to limit the wastes produced by the fish. The method used to determine digestibility can affect the value of the coefficients obtained (Cho et al., 1982). Thus, Lee (1997a) compared apparent nutrient digestibility of a diet by using chromic oxide indicator according to the various fecal collection methods (dissection, stripping or using fecal collection column attached to fish rearing tank), and suggested that stripping or fecal collection column could be a reliable procedure for measuring the nutrient digestibility in rockfish. Lee (1997a) also suggested that fecal collection column is more comfortable method than intestinal fecal collection in case of fry or juvenile rockfish because there is no clear anatomical distinction between intestinal regions and sufficient amount of feces may not be collected from smaller intestine. Lee (1997b) investigated the fatty acids digestibility of dietary lipid sources for rockfish. However, the digestibility of protein sources in rockfish feeds have not been studied. The present study, therefore, was conducted to determine apparent digestibility coefficients for dry matter, crude protein, energy and amino acids in white fish meal, anchovy meal, meat meal, feather meal, blood meal, soybean meal, corn gluten meal, cottonseed meal, wheat flour, and brewers yeast in diets for juvenile and grower rockfish using a fecal collector attached to fish rearing tank.

2. Materials and methods 2.1. Diet preparation The reference diet (Table 1) was formulated to satisfy the nutrient requirements of rockfish using white fish meal (produced by a steam dry method, Han Chang Fish Meal, Pusan, Korea) and squid liver oil (E-Wha Oil & Fat Ind., Pusan, Korea) based on our previous study (Lee and Lee, 1994a). Chromic oxide (Cr2O3) was used as an inert marker at a concentration of 0.5% in diet.

S.-M. Lee / Aquaculture 207 (2002) 7995 Table 1 Ingredients composition of the reference diet Ingredients White fish meal Wheat flour Brewers yeast Squid liver oilb Vitamin premixc Mineral premixd Alpha-cellulosee Carboxymethyl cellulosee Choline salte Chromic oxide
a b a

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Percentage 58.0 22.5 3.0 4.0 2.2 4.0 2.0 3.0 0.8 0.5

Produced by steam dry method. Provided by E-wha Oil & Fat Ind. Co., Pusan, Korea. c Vitamin premix contained the following amount which were diluted in cellulose (g/kg mix): L-ascorbic acid, 121.2; DL-a-tocopheryl acetate, 18.8; thiamin hydrochloride, 2.7; riboflavin, 9.1; pyridoxine hydrochloride, 1.8; niacin, 36.4; Ca-D-pantothenate, 12.7; myo-inositol, 181.8; D-biotin, 0.27; folic acid (98%), 0.68; p-aminobenzoic acid, 18.2; menadione, 1.8; retinyl acetate, 0.73; cholecalciferol, 0.003; cyanocobalamin, 0.003. d Mineral premix contained the following ingredients (g/kg mix): MgSO47H2O, 80.0; NaH2PO42H2O, 370.0; KCl, 130.0; Ferric citrate, 40.0; ZnSO47H2O, 20.0; Ca-lactate, 356.5; CuCl, 0.2; AlCl36H2O, 0.15; KI, 0.15; Na2Se2O3, 0.01; MnSO4H2O, 2.0; CoCl26H2O, 1.0. e Sigma, St. Louis, MO, USA.

Test ingredients for apparent digestibility were white fish meal (5-02-025), anchovy meal (5-01-985), meat meal (5-00-385), feather meal (5-03-795), blood meal (5-00-381), soybean meal (5-04-612), corn gluten meal (5-28-242), cottonseed meal (5-01-617), wheat flour (4-05-199), and brewers yeast (7-05-527). Ten test diets were formulated using 70% reference diet and 30% of each of the test ingredients on an air-dry basis as described by Cho and Slinger (1979). Proximate and amino acid composition of the test ingredients and diets are shown in Tables 2 and 3, respectively. Diets were mechanically mixed with distilled water (40 g/100 g diet mix), pressure-pelleted, and stored at 25 C until used. 2.2. Fish and experimental condition Juvenile rockfish (Sebastes schlegeli) obtained from the hatchery of National Fisheries Research and Development Institute (Pusan, Korea), were acclimated to the laboratory for 2 months (juvenile) and 10 months (grower), respectively. Afterwards, the experimental fish were randomly distributed into 200 l cylindrical fiberglass tanks (containing 100 l of water each) of a flow-through aquarium system (Fig. 1) and were hand-fed the reference diet (chromic oxide-free) to visual satiety twice daily and once a day for 10 weeks for juvenile and grower fish, respectively, based on a previous study (Lee et al., 2000). After the conditioning period, the initial weight (mean F S.E.) of 30 F 0.5 g and 300 F 4.2 g fish per tank were selected and redistributed into the same tanks with 50 and 8 fish per tank, respectively. Filtered seawater was supplied at a flow rate of 3 l/min to each rearing tank. Fish rearing tanks had a sloping bottom leading to a centrally located drainage slot, and the effluent water was directed first over a fecal collection column and then to waste. The water

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Table 2 Proximate and amino acids composition of the ingredients used to test diets Test ingredients WFMa AMb MMc FMd 91.8 2.7 0.4 2.8 2.2 5715 BMe 95.8 0.0 0.9 2.0 1.3 5889 SMf 53.5 1.3 4.7 6.6 33.9 4759 CGMg 75.3 2.9 0.6 3.2 18.0 5847 CSMh 39.8 0.6 22.3 5.9 31.4 4738 WFi 14.6 0.1 0.2 0.5 84.7 4433 Yeast j 49.3 0.0 0.1 6.2 44.3 4784

Proximate analysis (% in dry matter) Crude protein 72.5 73.8 84.3 Crude lipid 5.5 8.4 7.4 Crude fiber 0.6 0.8 3.0 Ash 19.4 16.3 5.6 NFEk 1.2 0.6 0.0 Gross energy 4898 5030 5485 (kcal/kg) Amino acids (% in protein) Arg 6.9 7.0 His 2.0 3.1 Ile 4.4 4.1 Leu 8.2 7.5 Lys 6.2 6.3 Met 2.3 2.0 Cys 1.2 1.2 Phe 4.3 4.1 Tyr 3.7 3.2 Thr 4.8 4.6 Val 4.9 4.6 Ala 6.5 7.5 Asp 9.9 9.4 Glu 14.9 14.4 Gly 6.9 9.3 Pro 6.8 6.0 Ser 5.1 4.4
a b c d e f g h i j k

7.6 1.6 2.6 5.9 4.2 1.4 1.3 3.2 2.3 3.2 4.1 8.5 7.2 12.2 16.7 13.3 4.1

6.0 0.8 4.2 7.5 1.8 0.8 3.0 4.6 2.5 4.1 5.5 4.4 5.9 10.0 6.9 22.8 8.5

3.9 5.7 0.4 14.3 7.6 0.9 0.9 8.1 2.6 4.7 8.5 9.1 11.0 8.8 4.7 3.2 5.2

7.5 2.3 4.6 8.2 5.0 0.4 1.5 5.4 3.6 4.2 4.5 4.6 11.3 18.6 4.5 8.0 5.3

3.1 1.5 3.7 16.1 1.3 1.6 1.5 6.0 4.9 3.3 3.9 8.4 5.7 19.3 2.5 11.4 4.9

12.0 2.5 3.4 6.7 3.6 1.0 1.6 6.1 3.0 3.7 4.6 4.5 9.7 21.3 4.6 6.1 4.8

4.1 2.2 3.5 7.0 2.0 0.4 1.9 4.9 2.7 2.9 4.1 3.2 4.2 32.7 3.7 14.1 4.8

5.1 2.1 4.8 7.8 5.0 0.4 1.8 5.1 3.6 5.1 5.7 7.8 9.0 17.1 5.1 8.9 5.6

White fish meal was produced by steam dry method. Anchovy meal from Chile supplied by Kum Sung Feed, Pusan, Korea. Meat meal was obtained from Agribrand Purina Korea, Seoul, Korea. Feather meal was obtained from Daehanfeed, Seoul, Korea. Blood meal was obtained from Kum Sung Feed. Soybean meal (dehulled, solvent extracted) was obtained from Kum Sung Feed. Corn gluten meal was obtained from Kum Sung Feed. Cottonseed meal (solvent extracted) was obtained from Kum Sung Feed. Wheat flour was obtained from Kum Sung Feed. Brewers yeast was obtained from Kum Sung Feed. Nitrogen-free extract was calculated by difference.

temperature was maintained at 15 F 0.2 C using water temperature control system (Koito, Japan). Photoperiod followed the natural conditions during the experimental period. 2.3. Fecal collection techniques Three replicate groups of fish were fed by hand the reference and test diets to visual satiety once a day at 15:00 h. Fish were carefully fed by the same person during each

S.-M. Lee / Aquaculture 207 (2002) 7995 Table 3 Proximate and amino acids composition of the reference and test diets fed to grower and juvenile rockfish Reference Test diets (70% reference + 30% ingredient) diet WFMa AMb MMc FMd BMe SMf CGMg CSMh WFi

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Yeast j

Proximate analysis (% in dry matter) Crude protein 47.5 55.4 55.6 59.0 59.2 62.9 48.9 56.0 45.4 38.0 48.5 Crude lipid 8.5 8.2 8.6 9.0 6.2 6.1 6.2 6.7 6.1 6.3 6.1 Crude fiber 3.6 2.6 3.3 4.2 2.7 2.9 3.8 2.3 8.8 2.8 2.4 Ash 15.4 17.1 16.0 12.9 11.9 11.1 12.7 11.3 12.8 11.2 12.6 NFEk 24.9 16.7 16.5 15.0 20.1 17.0 28.4 23.8 26.9 41.6 30.3 Gross energy 4742 4791 4757 5008 5023 5072 4703 5053 4741 4678 4768 (kcal/kg) n 3HUFAl 1.9 Amino acids (% in protein) Arg 6.7 6.9 His 2.1 1.9 Ile 4.4 4.5 Leu 8.2 8.5 Lys 6.9 5.9 Met 2.3 2.2 Cys 1.2 1.1 Phe 4.4 4.4 Tyr 3.4 3.5 Thr 4.8 4.7 Val 4.9 4.7 Ala 6.3 6.5 Asp 9.7 9.7 Glu 16.1 15.4 Gly 6.5 7.3 Pro 6.2 6.8 Ser 5.1 5.0
a b c d e f g h i j k l

6.9 2.4 4.6 8.5 6.1 2.1 1.2 4.5 3.5 4.8 4.8 6.8 9.7 15.5 7.2 5.6 4.8

7.2 1.7 3.7 7.4 5.1 1.8 1.1 4.0 2.9 4.1 4.5 7.4 8.6 14.4 10.9 9.9 4.6

5.9 1.3 4.0 7.4 3.7 1.4 1.9 4.2 2.9 4.1 4.7 5.1 7.3 12.1 6.2 20.9 6.3

5.5 3.5 2.5 11.3 6.1 2.0 1.0 6.2 3.1 4.7 5.9 7.8 10.0 12.4 5.9 6.4 5.0

7.0 2.0 4.5 8.3 5.6 1.3 1.3 4.7 3.5 4.6 4.7 5.9 10.2 16.8 6.1 7.4 5.1

5.1 1.7 4.1 12.0 3.9 2.0 1.3 5.1 4.1 4.0 4.4 7.4 7.8 17.2 4.8 9.2 4.9

7.8 2.1 4.2 8.0 5.4 1.7 1.2 4.8 3.2 4.5 4.8 6.0 9.6 17.3 6.3 7.2 5.0

6.4 2.1 4.4 8.3 5.7 2.0 1.3 4.5 3.2 4.6 4.8 6.1 9.2 18.3 6.4 6.2 5.1

6.2 2.0 4.5 8.2 5.6 1.8 1.3 4.6 3.4 4.9 5.0 6.9 9.4 16.2 6.4 7.4 5.2

White fish meal was produced by steam dry method. Anchovy meal from Chile supplied by Kum Sung Feed. Meat meal was obtained from Agribrand Purina Korea. Feather meal was obtained from Daehanfeed. Blood meal was obtained from Kum Sung Feed. Soybean meal (dehulled, solvent extracted) was obtained from Kum Sung Feed. Corn gluten meal was obtained from Kum Sung Feed. Cottonseed meal (solvent extracted) was obtained from Kum Sung Feed. Wheat flour was obtained from Kum Sung Feed. Brewers yeast was obtained from Kum Sung Feed. Nitrogen-free extract was calculated by difference. Highly unsaturated fatty acids (C ! 20), contained 0.7% EPA and 1.1% DHA in the reference diet.

feeding trial. Two hours after feeding, the rearing tank and collection column were brushed out to remove uneaten feed and fecal residues. The next day, feces were collected from the fecal collection columns at 08:00 and 13:00 h for 12 days. Feces collected from the settling columns were immediately filtered with filter papers (Whatman #1) for 60 min at 4 C and stored at 75 C for chemical analyses. Fecal samples from each tank were pooled at the end of experiment.

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Fig. 1. Digestibility tank system, a 150-mm diameter stand pipe was used to prevent water loss when fecal collection column was cleaning.

2.4. Analytical methods Freeze-dried feed and feces were finely ground using a grinder. Fish scales contaminating the feces collected were removed using a 300-mm sieve before analysis. Crude protein was determined by the Kjeldahl method using an Auto Kjeldahl System (Buchi B324/435/412, Switzerland). Crude lipid was determined by the ether-extraction method. Moisture was determined by oven drying at 105 C for 24 h. Crude fiber was determined by an automatic analyzer (Fibertec, Tecator, Sweden) and ash was determined by a muffler furnace at 550 C for 4 h. Nitrogen-free extract (NFE) was calculated by difference. Gross energy contents were analyzed using an adiabatic bomb calorimeter (Parr, USA). Amino acids were analyzed using an automatic analyzer (Hitachi Model 835-50, Japan) with an ion exchange column (Hitachi Resin #2619, 2.6 150 mm, Japan). Chromic oxide was determined by a wet-acid digestion method (Furukawa and Tsukahara, 1966). All chemical analyses (from each tank) were done in duplicate or triplicate. Apparent digestibility coefficients (ADC) for dry matter, nutrient and energy of the diets were determined using the following equations ADC of dry matter % 100 dietary Cr2 O3 =fecal Cr2 O3 100,

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ADC of nutrients or energy %   dietary Cr2 O3 fecal nutrient or energy 1 100: dietary nutrient or energy fecal Cr2 O3

The ADC for dry matter, crude protein, energy and amino acids were calculated from the respective digestibility coefficients for reference and test diets on the basis of the 30% substitution of test ingredient in the reference diet (Cho et al., 1982) ADC of test ingredient % 100=30 ADC in test diet 0:7 ADC in reference diet: 2.5. Statistical analyses All data were subjected to one-way analysis of variance, followed by Duncans multiple range test (Duncan, 1955) at a significance level of P < 0.01. The significances of difference in digestibility coefficients as influenced by the fish size ( juvenile and grower) were assessed by independent-samples t-test. Linear correlations were determined between nutrient digestibility and contents of test ingredients. All statistical analyses were carried out using the SPSS Version 7.5 (SPSS, Michigan Avenue, Chicago, IL, USA).

3. Results Apparent dry matter, crude protein and gross energy digestibility coefficients of test ingredients for rockfish are shown in Table 4. Apparent digestibilities of dry matter (32 95%), crude protein (63 95%) and energy (39 99%) were affected by test ingredients ( P < 0.01). Dry matter digestibility of animal products (range of 73 95%) was not significantly ( P > 0.01) different in the same fish size and this was less variable than that of plant products (range of 32 87%). Dry matter digestibility was more (> 73%) digestible in the high-protein ingredients (animal products and corn gluten meal) and less ( < 54%) digestible in the high nitrogen-free extract or fiber ingredients (soybean meal, cottonseed meal, wheat flour and/or brewers yeast) for this species. Dry matter digestibilities of white fish meal, anchovy meal, meat meal, feather meal, blood meal and corn gluten meal were significantly higher ( P < 0.01) than those of cottonseed meal, wheat flour, soybean meal and/or brewers yeast, however this value was not influenced by fish size in each test ingredient ( P > 0.05). Dry matter digestibility was significantly correlated with fiber (r = 0.70, P < 0.01 and r = 0.65, P < 0.05 in grower and juvenile fish, respectively), nitrogen-free extract (r = 0.88, P < 0.01 and r = 0.90, P < 0.01) and protein (r = 0.92, P < 0.01 for both) of the test ingredients, whereas this value was not correlated with ash (r = 0.45 and r = 0.53; P > 0.05).

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Table 4 Apparent digestibility coefficients (%) for dry matter, crude protein and energy of the test ingredients in grower and juvenile rockfisha Ingredients Dry matter Grower Juvenile White fish meal Anchovy meal Meat meal Feather meal Blood meal Soybean meal Corn gluten meal Cottonseed meal Wheat flour Brewers yeast SEMc 73 bcd 82 bcd 90 d 77 bcd 84 cd 59 bc 87 d 34 a 32 a 59 b 5.51 89 b 83 b 95 b 87 b 87 b 54 a 85 b 46 a 42 a 56 a 6.17 0.3 0.9 0.4 0.2 0.4 0.5 0.8 0.3 0.3 0.8 P-valueb Protein Grower Juvenile 88 bc 92 c 90 c 63 a 87 bc 84 bc 92 c 78 b 91 c 78 b 2.42 95 d 95 d 91 cd 79 ab 86 bc 80 ab 92 cd 81 ab 95 d 73 a 1.92 0.05 0.3 0.8 0.03 0.7 0.08 0.8 0.5 0.1 0.3 P-valueb Energy Grower Juvenile 90 c 93 c 90 c 73 b 86 c 64 b 89 c 41 a 39 a 66 b 3.04 99 d 96 cd 93 cd 85 c 84 c 61 b 89 cd 55 ab 46 a 60 b 3.17 0.2 0.6 0.4 0.04 0.5 0.8 0.9 0.06 0.2 0.4 P-valueb

a Values (mean of three replicate groups) in each column with a different letter are significantly different ( P < 0.01). b t-Test between fish size. c Standard error of the treatment mean calculated from the residual mean square in the analysis of variance.

Protein digestibilities of animal and plant products ranged 63 95% and 73 92%, respectively. Protein digestibilities of grower fish were significantly higher ( P < 0.01) ranging from 84% to 92% in white fish meal, anchovy meal, meat meal, blood meal, soybean meal, corn gluten meal and wheat flour, and those of juvenile were significantly higher ( P < 0.01) ranging from 91% to 95% in white fish meal, anchovy meal, meat meal and corn gluten meal. Protein digestibility of feather meal in grower fish was significantly ( P < 0.01) the lowest (63%), and this value of brewers yeast in juvenile fish was significantly ( P < 0.01) the lowest (73%). Protein digestibilities of ingredients for juvenile fish had relatively higher trend for high-protein ingredients, especially those of white fish meal and feather meal in juvenile fish had higher ( P < 0.05) than grower fish. Protein digestibility was not correlated with ash, fiber, nitrogen-free extract or protein content of test ingredients (r > 0.5, P > 0.05 for both fish size). Differences in energy digestibility reflected differences in dry matter digestibility. Energy digestibilities of animal and plant products ranged 73 99% and 39 89%, respectively. Energy digestibilities of grower fish were significantly higher ( P < 0.01) in white fish meal, anchovy meal, meat meal, blood meal and corn gluten meal than other ingredients, and those of juvenile were significantly higher ( P < 0.01) in white fish meal, anchovy meal, meat meal and corn gluten meal than soybean meal, cottonseed meal, wheat flour and brewers yeast. Energy digestibilities of cottonseed meal and wheat flour were low ranging from 39% to 41% in grower fish and from 46% to 55% in juvenile fish; significantly lowest ( P < 0.01) among the ingredients tested. Energy digestibility was significantly correlated with fiber (r = 0.73, P < 0.01 and r = 0.63, P < 0.05 in grower and juvenile fish, respectively), nitrogen-free extract (r = 0.90, P < 0.01 and r = 0.93, P < 0.01), protein (r = 0.91, P < 0.01 for both) and ash (r = 0.66, P < 0.05 and r = 0.72, P > 0.01). Energy digestibilities of rockfish were positively correlated with protein content of

Table 5 Apparent amino acid availabilities (%) of the ingredients in grower rockfisha Ingredientsb Amino acids Arg WFM AM MM FM BM SM CGM CSM WF Yeast SEMc 93 b 93 b 95 b 74 a 90 b 88 b 92 b 86 b 94 b 87 b 2.13 His 87 ns 88 79 65 90 74 83 68 79 76 5.68 Ile 92 c 89 bc 90 c 71 ab 65 a 83 abc 93 c 78 abc 91 c 76 abc 4.06 Leu 93 bc 98 c 91 bc 68 a 99 c 86 bc 99 c 83 b 92 bc 81 ab 3.28 Lys 90 b 90 b 90 b 76 a 91 b 88 b 87 b 77 a 91 b 80 ab 2.45 Met 99 ns 98 99 96 98 90 99 82 97 83 4.43 Cys 75 cd 94 d 64 bc 44 a 48 ab 71 bcd 80 cd 63 bc 86 cd 86 cd 5.56 Phe 93 bcd 95 cd 91 bcd 71 a 98 d 84 bc 97 cd 86 bcd 92 bcd 81 ab 2.91 Tyr 92 ns 93 92 85 87 84 97 87 95 87 5.06 Thr 91 cd 96 d 89 cd 66 a 89 cd 86 bcd 93 cd 80 bc 95 d 74 ab 3.18 Val 92 cd 96 cd 89 cd 66 a 99 d 87 bcd 91 cd 82 bc 94 cd 73 ab 3.32 Ala 94 b 93 b 95 b 73 a 95 b 87 a 97 b 87 a 93 b 83 a 4.45 Asp 86 bc 91 c 81 bc 64 a 88 c 82 bc 87 c 81 bc 85 bc 75 b 2.51 Glu 91 bcd 91 bcd 88 bcd 69 a 86 bcd 86 bcd 94 cd 84 bc 95 d 83 b 2.27 Gly 82 b 85 b 88 b 62 a 81 b 82 b 79 b 78 b 85 b 77 ab 3.75 Pro 89 bc 90 bcd 84 b 69 a 90 bcd 90 bcd 97 cd 89 bc 99 d 86 b 2.43 Ser 90 c 90 c 89 c 61 a 83 bc 84 bc 91 c 77 bc 91 c 69 ab 3.55 S.-M. Lee / Aquaculture 207 (2002) 7995

ns, Not significant ( P > 0.01). a Values (mean of three replicate groups) in each column with a different letter are significantly different ( P < 0.01). b See Table 2. c Standard error of the treatment mean calculated from the residual mean square in the analysis of variance.

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Table 6 Apparent amino acid availabilities (%) of the ingredients in juvenile rockfisha Ingredientsb Amino acids S.-M. Lee / Aquaculture 207 (2002) 7995 Arg WFM AM MM FM BM SM CGM CSM WF Yeast SEMc 99 d 98 d 92 bcd 88 abc 91 abc 86 ab 89 bcd 87 abc 96 cd 81 a 2.23 His 99 b 97 b 90 b 77 ab 99 b 81 ab 90 b 81 ab 98 b 63 a 5.79 Ile 98 c 96 c 81 ab 84 abc 77 a 76 a 95 bc 80 ab 84 ab 72 a 3.70 Leu 99 d 99 d 85 abc 84 abc 96 cd 80 ab 99 d 85 abc 92 bcd 74 a 3.25 Lys 97 c 95 c 89 bc 78 ab 86 bc 82 abc 84 abc 79 ab 95 c 70 a 3.26 Met 100 c 99 bc 94 bc 83 abc 95 bc 71 ab 98 bc 70 ab 94 bc 61 a 6.24 Cys 95 ns 93 75 71 81 76 80 63 92 76 8.60 Phe 98 c 97 c 83 ab 85 abc 96 bc 78 a 96 bc 86 abc 97 c 74 a 3.06 Tyr 99 ns 99 88 87 92 85 80 87 99 73 6.38 Thr 100 d 98 cd 83 abc 84 abc 93 bcd 81 ab 88 bcd 81 ab 91 bcd 71 a 3.60 Val 100 c 97 bc 82 ab 89 abc 98 bc 83 abc 92 bc 82 ab 94 bc 70 a 4.47 Ala 100 d 97 d 91 bcd 85 bc 94 bcd 83 ab 95 cd 85 bc 90 bcd 74 a 2.47 Asp 99 d 94 cd 82 abc 82 abc 93 bcd 78 ab 87 bcd 82 abc 88 bcd 72 a 3.31 Glu 99 d 97 cd 88 abc 85 ab 88 abc 82 a 95 bcd 85 ab 96 cd 79 a 2.37 Gly 99 ns 97 93 85 87 82 84 83 91 86 4.34 Pro 99 b 93 b 62 a 94 b 94 b 86 ab 98 b 93 b 97 b 82 ab 6.94 Ser 100 b 96 b 75 ab 87 ab 93 b 80 ab 92 b 83 ab 92 b 64 a 5.37

ns, Not significant ( P > 0.01). a Values (mean of three replicate groups) in each column with a different letter are significantly different ( P < 0.01). b See Table 2. c Standard error of the treatment mean calculated from the residual mean square in the analysis of variance.

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ingredients, except for feather meal. Energy digestibility of feather meal in juvenile fish was significantly higher ( P < 0.05) than that in grower fish, however energy digestibilities of other ingredients were not affected by fish size ( P > 0.05). Apparent amino acid availability coefficients of test ingredients for grower and juvenile rockfish are shown in Tables 5 and 6, respectively. Amino acid availability values for the test ingredients followed similar trend to values of protein digestibility. Amino acid availability values, except for histidine, methionine and tyrosine, in white fish meal, anchovy meal, meat meal, corn gluten meal and wheat flour for grower fish were slightly or significantly ( P < 0.01) higher than other test ingredients. Amino acids availabilities of feather meal in grower fish were the lowest among test ingredients with the exception of isoleucine, methionine and tyrosine, and the availabilities of isoleucine from blood meal, methionine from cottonseed meal and tyrosine from soybean meal were the lowest. Amino acid availabilities for white fish meal, anchovy meal and corn gluten meal in juvenile fish were slightly or significantly ( P < 0.01) higher than those of other test ingredients with the exception of cystine, tyrosine and glycine. Amino acid availability values of brewers yeast in juvenile fish were the lowest among test ingredients with the exception of cystine, glycine and proline, and those of cystine and glycine from cottonseed meal and proline from meat meal were the lowest. Amino acid availability values of animal protein sources such as fish meals and feather meal were relatively high, but those of plant protein source such as soybean meal and brewers yeast were lower in juvenile fish than in grower fish.

4. Discussion The digestibility coefficients of feed ingredients provide insight concerning nutrient utilization and should enable more accurate ingredient substitutions in diets designed for target fish species. The nutrients digestibility will vary depending on the composition of ingredients used. It has been reported that carnivorous fish tend to utilize the dry matter and energy in animal products better than that in plant products (Cho et al., 1982; Bergot and Breque, 1983; Sullivan and Reigh, 1995). The low dry matter and energy digestibility of some plant products appears to be related to the quantity and chemical composition of the carbohydrates they contain. Dry matter and energy digestibilities of rockfish were also negatively correlated with carbohydrate content of ingredients tested. Similar trends on the inverse relationship between carbohydrate content and dry matter or energy digestibility have been reported for salmonids (Sugiura et al., 1998), hybrid striped bass (Sullivan and Reigh, 1995) and red drum (McGoogan and Reigh, 1996). However, dry matter and energy digestibilities of corn gluten meal were as high as those of fish meals despite its 18% carbohydrate (Table 2) regardless of fish size. These results indicate that digestibility of carbohydrate for rockfish could vary depending on plant products and their carbohydrate level, and plant products containing the proper ratio of protein and carbohydrate should be selected carefully to maximize the availability of carbohydrate energy in rockfish diets. Dietary fiber is not utilized by fish (NRC, 1993) and thus it may cause lower dry matter and energy digestibilities of fish. Other studies have observed a negative cor-

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relation between fiber content of the diet and dry matter or energy digestibility of fish (Hilton et al., 1983; Kirchgessner et al., 1986). Dry matter and energy digestibilities in this study also tended to decline as the fiber content of ingredients increased. Especially, dry matter and energy digestibilities of cottonseed meal and wheat flour were significantly lower than those of animal products, and this trend may be due to high content of fiber or nitrogen-free extract in these ingredients. However, protein digestibility was not correlated with fiber content of ingredient. Low levels of dietary fiber ( < 10%) did not have any negative effects on growth and feed efficiency for this species fed isocaloric diets (Lee and Lee, 1994b). High dietary fiber levels can reduce the utilization of other nutrients (Walker, 1975; Shah et al., 1982; Hilton et al., 1983; Anderson et al., 1984). Low protein digestibility in some fish fed the high fiber diets has been attributed to decreased proteolytic enzyme activity (Falge et al., 1978) and shortened gut-transit time (Jobling, 1981; Steffens, 1989) that results in incomplete digestion and absorption. Generally, the protein quality of dietary ingredients is the leading factor affecting fish performance, and protein digestibility is the first measure of its availability by fish. Another study with rockfish has indicated that high quality fish meal supports optimal growth, while other protein feedstuffs can lead to sub-optimal fish performance (Lee et al., 1996). Protein from fish meal was well digested by most carnivorous fish (Willson and Poe, 1985; McGoogan and Reigh, 1996; Sugiura et al., 1998), this is consistent with results of this study. The protein of meat meal, corn gluten meal and wheat flour was well digested in this study, this indicates that each of these ingredients can be used efficiently as a partial protein source for rockfish diets. The relatively low protein digestibility of feather meal could be explained partially by indigestible nitrogen compounds in the ingredient. Protein digestibility of blood meal in rockfish was higher than that in chinook salmon (Hajen et al., 1993), but lower than that in red drum (McGoogan and Reigh, 1996). Some results showed that protein digestibility of blood meal were variable (Cho and Slinger, 1979; Smith et al., 1980; Cho et al., 1982; Asgard and Austreng, 1986; Bureau et al., 1999). Differences in digestibility of blood meal among studies may be due to differences in fish species, meal processing condition or meal quality. Protein digestibility of soybean meal in this study was very similar to that for red drum and hybrid striped bass (Sullivan and Reigh, 1995; Gaylord and Gatlin, 1996). Soybean meal is an important protein source as a partial substitute for fish meal in rockfish diets (Lee et al., 1996). The higher soybean meal utilization with fish size showed in Lee and Jeons (1996) study agree to this study showing that protein digestibility of soybean meal was relatively improved in grower fish. Low protein digestibility of cottonseed meal may be due to the higher content of fiber (Jones, 1979). Yeast products such as brewers yeast are frequently used as feed additive in aquaculture because they contain essential nutrients and may also contain unknown growth factor, attractant, etc. However, the use of yeast as feed ingredients for animals is poor due to yeast cell walls being composed of complex heteropolysaccharides in a structured carbohydrate protein complex such as mannoprotein and glucan (Johnson et al., 1980; Farkas, 1985). The low protein digestibility of brewers yeast in this study may be related to the rough cell wall of yeast. McGoogan and Reigh (1996) reported that high protein digestibility for red drum might be related to high protein content and low carbohydrate of ingredients. However, protein

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digestibility in this study was not correlated with protein and non-protein contents of ingredients tested for both fish size. For example, protein digestibility of feather meal which contained high protein (92%) was relatively low and that of wheat flour which contained low protein (15%) was relatively high compared to that of some other ingredients. Differences in protein digestibility among studies may be due to different experimental conditions such as fish species, digestible protein compounds and processing treatments of ingredients. Protein quality of dietary protein sources depends on the amino acid composition and their availability. Deficiency of an essential amino acid leads to poor utilization of the dietary protein and consequently reduces growth and decreases feed efficiency. Although the data presented in this study suggest a reasonable agreement between protein and amino acid digestibilities, individual amino acid availabilities within a feed ingredient are variable. Since essential amino acids requirements and their availabilities of different protein sources have not been reported for rockfish, use of amino acid availability data obtained from this study should allow for more accurate and economical rockfish feed formulation. Anderson et al. (1992) suggested that use of true amino acid availability would allow more accurate and economical feed formulation than use of apparent amino acid availability for Atlantic salmon when the feces were collected by stripping of anal region. The apparent protein and amino acid digestibilities were known to be influenced by metabolic fecal nitrogen excretion (Ogino and Chen, 1973; Willson et al., 1981). Feces collected manually from intestine may have more digestive juice and mucus by handling stress than did collection from collection column, thus increasing the endogenous nitrogen material and causing an underestimation of digestibility. In considering the amount of endogenous nitrogen materials such as bile, enzymes, epithelial cells and mucus, true nutrient digestibility appeared to be more desirable in manual fecal collection comparing to fecal collection column. When fecal collection column methods are employed, sufficient feces could be collected with the same feeding conditions without fish-handling, thus endogenous nitrogen material produced by stress might be decreased. Lee (1997a) investigated nutrient digestibility using reference diet and protein-free diet, and suggested that amino acid availability would not be greatly influenced by metabolic fecal nitrogen for rockfish when the feces were collected by fecal collection column as employed in this study. Rychly and Spannhof (1979) also showed that true protein digestibility of rainbow trout was slightly higher than apparent protein digestibility, but the difference was negligible when feces were collected from the bottom of the aquaria. Digestible energy requirements of fish range from 2.33 to 4.10 kcal/g diet depending on species and dietary protein levels (NRC, 1993). Digestible energy values of test ingredients for rockfish and other fish species are shown in Table 7. Digestible energy values of the different feedstuffs varied among species. Digestible energy values of 4.7 5.2 kcal/g for fish meals, meat meal, blood meal and corn gluten meal in the present study were higher than those for rainbow trout, and the values (2.9 3.0 kcal/g) of soybean meal were same to those (2.9 3.0 kcal/g) determined for rainbow trout and red drum (NRC, 1993; Gaylord and Gatlin, 1996). Digestible energy values of cottonseed meal and brewers yeast in the present study were lower than those for rainbow trout, channel

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Table 7 Digestible energy values (kcal/kg dry matter) of ingredients for rockfish and other fish species Ingredients Rockfisha Grower White fish meal Anchovy meal Meat meal Feather meal Blood meal Soybean meal Corn gluten meal Cottonseed meal Wheat flour Brewers yeast
a b c

Juvenile 4886 4823 5097 4865 4966 2921 5203 2612 2051 2857

Average 4655 4757 5025 4529 5013 2979 5211 2273 1897 3009

Rainbow troutb 3490 4570

Channel catfishb

Chinook salmonb

Red drumc

4425 4692 4953 4193 5060 3037 5219 1934 1744 3161

5248

4611 2934 4681 2689 1805 3787

3667

2004 3250

3005 3272

3380

Data from the current study. Data reported in NRC (1993). Gaylord and Gatlin (1996).

catfish and/or red drum (NRC, 1993; Gaylord and Gatlin, 1996). From data obtained in the present study with rockfish, energy was readily obtainable from animal products and corn gluten meal containing highly digestible protein and lipid contents with the exception of feather meal. Lipid and carbohydrate are important energy sources, but the relative amounts of carbohydrate energy in animal products such as fish meal and lipid energy in plant products such as wheat flour used in this study (Table 2) were low. Although the data of lipid and carbohydrate digestibility determined in this study were not shown in the table, lipid digestibility of fish meals in the present study was determined to be 93 98%, which is similar to lipid digestibility (95 97%) of fish oil and fish meal for rockfish (Lee, 1997b) and rainbow trout (Cho et al., 1982). This indicates that carnivorous fish including rockfish can easily utilize the fish oil for growth or energy as described by Sargent et al. (1989). The ability of fishes to utilize carbohydrate as an energy source differs among species and appears to be rather limited in carnivorous fish (Wilson, 1994). In this study, it appeared that energy from plant products containing relatively high carbohydrate content was not readily available to rockfish. Energy digestibility in this study was negatively related to the carbohydrate content and positively related to protein content of test ingredients. The nitrogen-free extract digestibilities of soybean meal (56% and 42% in grower and juvenile fish, respectively), cottonseed meal (15% and 21%), wheat flour (24% and 23%) and brewers yeast (76% and 54%) which had the relatively low energy digestibility values were low for rockfish. Rainbow trout are capable of digesting only 38 55% of dietary starch (Bergot and Breque, 1983), whereas common carp can digest up to 85% of the starch in a high-starch diet (Chiou and Ogino, 1985). In considering these results and strong carnivorous feeding habit of rockfish (Hatanaka and Iizuka, 1962), the major factors affecting digestible energy of feed ingredients for rockfish appear to be the composition of protein and lipid.

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In summary, white fish meal, anchovy meal, meat meal, and corn gluten meal showed the highest crude protein and energy digestibilities among ingredients tested. Dry matter and energy digestibilities of cottonseed meal and wheat flour were lower than those of other ingredients, probably due to the high fiber and/or nitrogen-free extract contents. Dietary protein sources for rockfish should be selected carefully on the basis of amino acid balance and price, and its energy sources should be considered on the product origin, processing procedure, chemical composition, etc. This information may be useful for more precisely formulating diets for rockfish on the basis of available nutrients in order to minimize production costs as well as restrict waste production.

Acknowledgements This work was supported by the funds of the Ministry of Marine Affairs in Korea.

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