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한수지 47(6), 824-83, 204 Original Article Kor J Fish Aquat Sci 47(6),824-83,204 넙치 (Paralichthys olivaceus) 치어사료에서산가수분해농축대두박 (Acid-concentrated soybean meal) 의어분대체효과및소화율평가 김성삼 오대한 조성준 2 서상현 2 한현섭 이경준 3 * 국립수산과학원사료연구센터, CJ 제일제당생물자원연구소, 2 CJ 제일제당소재연구소, 3 제주대학교해양생명과학과 Evaluation of Acid-concentrated Soybean Meal as a Fishmeal Replacement and its Digestibility in Diets for Juvenile Olive Flounder Paralichthys olivaceus Sung-Sam Kim, Dae-Han Oh, Seong-Jun Cho 2, Sang-Hyun Seo 2, Hyon-Sob Han and Kyeong-Jun Lee 3 * Aquafeed Research Center, National Fisheries Research and Development Institute, Pohang 79-923, Korea Animal Bioscience Research Institute, CJ CheilJedang Corporation, Incheon 400-03, Korea 2 Food Ingredients R&D Center, CJ CheilJedang Corporation, Seoul 52-866, Korea 3 Department of Marine Life Sciences, Jeju National University, Jeju 690-756, Korea This study was conducted to investigate the effects of fishmeal replacement with acid-concentrated soybean meal (ACSBM) on growth performance, blood biochemistry, and ingredient digestibility in juvenile olive flounder Paralichthys olivaceus. Six experimental diets were formulated to replace fishmeal protein with ACSBM at 0%, 20%, 30%, 40%, 50%, and 60% (designated ACSBM0, ACSBM20, ACSBM30, ACSBM40, ACSBM50, and ACSBM60, respectively). Triplicate groups of fish (initial fish mean weight: 4.3±0.03 g) were fed the experimental diets to apparent satiation (twice daily at 08:00 and 8:00 h). After a 2-week feeding trial, a total of 80 healthy fish were randomly distributed into three Guelph system tanks at a density of 60 fish/tank (initial fish mean weight : 50.6±2.4 g) to test the apparent digestibility coefficients of the ingredients (ACSBM, fishmeal, and soybean meal). Although negative effects were observed with ACSBM40, ACSBM50 and ACSBM60 after 2 weeks of feeding, up to 20% of the fishmeal protein could be successfully replaced with ACSBM without significant growth depression. Hemoglobin and hematocrit values of fish fed the ACSBM50 and ACSBM60 diets were significantly lower than those of fish fed the ACSBM0 diet. Glucose values of fish fed the ACSBM60 diet were significantly higher than those of fish fed the ACSBM0 and ACSBM20 diets. Digestibility of protein in ACSBM and soybean meal was 85.9% and 82.5%, respectively. Results indicated that at least 20% of fishmeal protein can be replaced by ACSBM in diets of juvenile olive flounder without supplementation of limiting amino acids. Key words: Olive flounder, Fish meal, Acid-concentrated soybean meal, Digestibility, Replacement 서론,. (50-70%). (Yang et al., 20; Song et al., 204). http://dx.doi.org/0.5657/kfas.204.0824 Kor J Fish Aquat Sci 47(6) 824-83, December 204 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial Licens (http://creativecommons.org/licenses/by-nc/3.0/)which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. Received 22 October 204; Revised 4 November 204; Accepted 3 November 204 *Corresponding author: Tel: +82. 64. 754. 3423 Fax: +82. 64. 756. 3493 E-mail address: kjlee@jejunu.ac.kr Copyright 204 The Korean Society of Fisheries and Aquatic Science 824 pissn:0374-8, eissn:2287-885

산가수분해농축대두박의어분대체효과 825 (Storebakken et al., 2000; Leenhouwers et al., 2006; Gatlin et al., 2007). protease inhibitor, tannins, lectins non-starch polysaccharides (antinutritional factors, ANF) (Francis et al., 200). ANF, (bioprocess) (Rumsey et al., 994; Buttle et al., 200; Refstie et al., 998, 2005).. (soy protein isolates, SPI), (soy protein concentrates, SPC), (high protein soybean meal, HSB) (soy products) (Chou et al., 2004; Glencross et al., 2005). (Ao, 20; Cavins et al., 972). (acid-concentrated soybean meal, ACSBM) (defatted soybean meal). (steaming roasting) oligosaccharides, lectins, soya antigens, phytase trypsin inhibitors ANF (Refstie et al., 998; Watson et al., 204). in vivo,. 재료및방법 산가수분해농축대두박제조공정 ( 67.0%, 3.4%, 3.5%) CJ. 5. N HCl ph 4.5. 30 5,000 g 20, (ACSBM). 실험사료 ( 67.0%, 3.4%, 3.5%) CJ. Table. Formulation and proximate composition of the experimental diets for olive flounder Paralichthys olivaceus Ingredients Diets ACSBM0 ACSBM20 ACSBM30 ACSBM40 ACSBM50 ACSBM60 Fish meal 50.0 40.0 35.0 30.0 25.0 20.0 ACSBM 0.0 0.0 5.0 20.0 25.0 30.0 Soybean meal 6.0 6.0 6.0 6.0 6.0 6.0 Corn gluten meal 6.0 6.2 6.4 6.6 6.7 6.9 Wheat flour 25.5 24.7 24.0 23.4 23.0 22.4 Fish oil 0.0 0.6..5.8 2.2 Mineral mix 2.0.0.0.0.0.0 Vitamin mix 3.0.0.0.0.0.0 Choline chloride 0.5 0.5 0.5 0.5 0.5 0.5 Proximate composition Protein (% DM) 49.6 49.6 49. 49. 49.3 49.3 Lipid (% DM) 5.7 6. 5.3 6. 5.7 6.5 Ash (% DM) 9.5 8. 7.6 6.8 6.3 5.6 Acid-concentrated soybean meal (ACSBM) was provided by CJ CheilJedang Corp. 2 MgSO 4.7H 2 O, 80.0; NaH 2 PO 4.2H 2 O, 370.0; KCl, 30.0; Ferric citrate, 40.0; ZnSO 4.7H 2 O, 20.0; Ca-lactate, 356.5; CuCl, 0.2; AlCl 3.6H 2 O, 0.5; Na 2 Se 2 O 3, 0.0; MnSO 4.H 2 O, 2.0; CoCl 2.6H 2 O,.0. 3 L-ascorbic acid, 2.2; DL-αtocopheryl acetate, 8.8; thiamin hydrochloride, 2.7; riboflavin, 9.; pyridoxine hydrochloride,.8; niacin, 36.4; Ca- D -pantothenate, 2.7; myo-inositol, 8.8; D -biotin, 0.27; folic acid, 0.68; p-aminobezoic acid, 8.2; menadione,.8; retinyl acetate, 0.73; cholecalficerol, 0.003; cyanocobalamin, 0.003.

826 김성삼ㆍ오대한ㆍ조성준ㆍ서상현ㆍ한현섭ㆍ이경준 Table Table 2. ACSBM 20%, 30%, 40%, 50% 60% 6 (ACSBM0, AC- SBM20, ACSBM30, ACSBM40, ACSBM50 ACSBM60)., 30% (NVM-4-2P, Daeyoung, Korea),. (SMC-2, Kuposlice, Busan, Korea). 24 h -20 C. 실험어및사육관리. 2. ( : 4.3 0.03 g) 8 50 L 40. 2-3 L/min. (5-22 ) 2-h light/2-h dark 2 (08:00 h 8:00 h). 3 24 h. 2. 샘플수집및분석 (weight gain), (specific growth rate), (feed conversion ratio), (protein efficiency ratio) (survival). 3 2-phenoxyethanol (200 ppm). hematocrit, hemoglobin. alanine aminotransferase (ALT), aspartate aminotransferase (AST). Hematocrit microhematocrit technique, hemoglobin, ALT, AST, protein glucose (Express plus system, Bayer, USA). AOAC (995) (25, 3 h), (550, 6 h), (Kejltec system 2300, Table 2. Amino acid composition (% protein) of the experimental diets for olive flounder Paralichthys olivaceus Amino acids Diets ACSBM0 ACSBM20 ACSBM30 ACSBM40 ACSBM50 ACSBM60 Arginine 6.07 6.53 6.2 6.08 6.52 5.94 Histidine 2.45 2.72 2.75 2.52 2.83.25 Isoleucine 4.6 4.97 4.7 4.77 5.27 4.44 Leucine 8.08 7.8 7.32 7.63 7.43 8.56 Lysine 5.40 5.00 5.06 4.7 4.42 5.59 Methionine 0.94 0.76 0.68 0.6 0.53 0.48 Phenylalanine 4.72 5.36 5.5 5. 5.83 4.99 Threonine 4.60 4.72 4.62 4.46 4.84 3.67 Valine 5.02 4.95 4.6 4.77 4.72 4.73 Aspartic acid 9.83 9.30 9.02 9.7 9.44.0 Glutamic acid 3.9 2.9 2.8 3.4 3. 7.7 Serine 4.9 4.6 4.69 4.97 4.85 5.37 Proline 2.6 3.7 7.5 6.4 5.6 2.2 Glycine 6.22 5.78 5.30 5.22 5.3 5. Alanine 7.07 6.67 6.9 6.26 6.03 5.78 Tyrosine 2.69 2.98 2.82 2.75 3.24 2.68 Cystine 0.86.3 0.55.7 0.2 0.53

산가수분해농축대두박의어분대체효과 827 Sweden), Folch et al. (959) Soxhlet (Soxhlet heater system C-SH6, Korea). Sycom S-433D automatic amino acid analyzer (Sykam, Eresing, Germany). 사료원료의외관상소화율측정 (,, ) (reference diet) (test diets). Table 5. 7:3. (indicator) chromium oxide (Cr 2 O 3, Sigma-Aldrich).0%. 2 (Guelph system). ( : 50.6 2.4 g) 6 400 L Guelph system 60. 2 L/min. 7:00 h. 09:00 h (feces). (0 mm, F093-0, Chmlab) -70 C. 0 (9-22 ). chromium oxide Divakaran et al. (2002). (550 ) 3 h. chromium oxide mono-chromate 5-0 mg glass test tube. glass test tub perchloric reagent 4 ml. Perchloric reagent 00 ml 200 ml, 70% perchloric acid 200 ml. perchloric reagent glass test tube 300 20. 50 ml 3 25 ml. (Beckman DU-730, USA) 350 nm. Standard standard Chromium oxide.. 참고사료및실험사료의영양소소화율 (%) =00 [ ( %/ %) ( %/ %)] (%) =[( ) (0.7 )]/[(- 0.7) ] (Completely randomized design), SPSS (Version 2.0) One-way ANOVA. Tukey's test (P 0.05) (mean SD). arcsine. 결과및고찰 2 Table 3. 20% 30% ACSBM20, ACSBM30 ACSBMS0, 40% ACSBM40, ACSBM50 ACSBM60 ACSBM0.. ACSBM20 ACSBM0 ACSBM30, AC- SBM40, ACSBM50, ACSBM60 ACSBM0. AC- SBM20 ACSBM30, ACSBM40, ACSBM50, ACSBM60 ACSBM20.., 40%. 30% 30%. 20-30% (Chou et al., 2004; Hernandez et al., 2007; Lim and Lee, 2008). Soy protein hydrolysates 85% (Song et al., 204), Soy protein concentrate (SPC) (Atlantic halibut, Berge et al.,999), (Atlantic salmon, Refstie et al.,200), (Common carp, Escaffre et al., 997), (Rainbow trout, Médale et al., 998; Mambrini et al.,

828 김성삼ㆍ오대한ㆍ조성준ㆍ서상현ㆍ한현섭ㆍ이경준 Table 3. Growth performance of olive flounder Paralichthys olivaceus fed the six experimental diets for 2 weeks Diets ACSBM0 ACSBM20 ACSBM30 ACSBM40 ACSBM50 ACSBM60 IBW (g) 2 4.2±0. 4.3±0.0 4.2±0.0 4.3±0.0 4.3±0. 4.2±0.0 FBW (g) 3 49.7±0.5 c 49.4±. c 45.6±.7 bc 44.7±0.9 b 40.0±3.3 a 36.±0.9 a WG (%) 4 249±3 c 246±8 bc 222±2 bc 23±6 b 80±24 a 53±7 a FCR 5.35±0.03 c.36±0.0 bc.42±0.08 ab.45±0.04 ab.7±0.20 a.97±0. a PER 6.48±0.04 c.49±0. c.44±0.08 c.40±0.04 bc.20±0.4 ab.03±0.06 a SGR (%) 7.69±0.0 b.66±0.02 b.6±0.0 b.54±0.02 b.39±0. a.25±0.04 a FI (g) 8 47.9±0.7 b 47.7±2.4 b 44.5±.6 ab 44.0±0.3 a 43.4±0.6 a 42.8±0.6 a Survival (%) 95.8±.4 97.5±0.0 95.8±3.8 95.0±2.5 93.3±6.3 89.2±5.2 Mean values of triplicate groups, values are presented as mean ± SD. Values in the same row having different superscript letters are significantly different (P<0.05). 2 Initial mean body weight. 3 Final mean body weight. 4 Weight gain (%)=00 (final mean body weight initial mean body weight)/initial mean body weight. 5 Feed conversion ratio=dry feed fed/wet weight gain. 6 Protein efficiency ratio=wet weight gain/total protein given. 7 Specific growth rate (%)=[(log e final body weight log e initial body weight)/days] 00. 8 Feed intake (g) = dry feed fed/fish. 999) (Senegalese sole, Aragão et al., 2003) (40-75%). Turbot SPC 25% (Day et al., 2000; Deng et al., 2006). (methionine, lysine), (Deng et al., 2006). (methionine) methionine (Table 2). 20%. 20%, (methionine, lysine ),,, phytase 20%. Table 4. Hematocrit AC- SBM20, ACSBM30 ACSBM40 ACSBM0 ACSBM50 AC- SBM60 ASBM0. Hemoglobin ACSBM20 ACSBM30 ACSBM0, ACSBM40, ACSBM50 ACSBM60 ACSBM0. Glucose ACSBM60 ACSBM0 ACSBM20. ALT, AST, protein.. ACSBM hematocrit, hemoglobin, glucose. ACSBM Table 4. Serological characteristics of olive flounder Paralichthys olivaceus fed the six experimental diets for 2 weeks Diets ACSBM0 ACSBM20 ACSBM30 ACSBM40 ACSBM50 ACSBM60 Hematocrit (%) 2.3±0.6 c 20.±.2 c 2.3±2. c 9.±0.2 bc 6.8±0.7 ab 5.7±.2 a Hemoglobin (g/dl) 4.54±0.03 c 4.60±0.20 c 4.46±0.29 c 4.29±0.35 bc 3.74±0.2 ab 3.52±0.35 a ALT (U/L) 32.9±5.5 34.3±3.6 4.0±8.4 38.2±7.7 34.0±8.7 29.7±6.4 AST (U/L).5±.8 3.0±.4 2.6±. 0.±.5 0.5±.3 0.7±3.3 Protein (g/dl) 4.23±0.2 3.76±4.08 4.08±0.46 4.20±0.9 4.76±0.25 4.49±0.23 Glucose (mg/dl) 8.5±0.7 a 8.7±.5 a 22.8±4.7 ab 22.6±2.6 ab 26.8±3.5 ab 29.5±6.6 b Mean values of triplicate groups, values are presented as mean ± SD. Values in the same row having different superscript letters are significantly different (P< 0.05).

산가수분해농축대두박의어분대체효과 829 Table 5. Formulation of the reference diet for digestibility test (% of DM basis) Ingredients Contents White fish meal 42.5 Soybean meal 6.0 Corn gluten meal 6.0 Wheat flour 30.0 Mineral mix.0 Vitamin mix 2.0 Yeast.0 Fish oil 0.0 Choline chloride 0.5 CMC.0 Chromium oxide 3.0 Test ingredient 0 MgSO 4.7H 2 O, 80.0; NaH 2 PO 4.2H 2 O, 370.0; KCl, 30.0; Ferric citrate, 40.0; ZnSO 4.7H 2 O, 20.0; Ca-lactate, 356.5; CuCl, 0.2; AlCl 3.6H 2 O, 0.5; Na 2 Se 2 O 3, 0.0; MnSO 4.H 2 O, 2.0; CoCl 2.6H 2 O,.0. 2 L-ascorbic acid, 2.2; DL-αtocopheryl acetate, 8.8; thiamin hydrochloride, 2.7; riboflavin, 9.; pyridoxine hydrochloride,.8; niacin, 36.4; Ca- D -pantothenate, 2.7; myo-inositol, 8.8; D -biotin, 0.27; folic acid, 0.68; p-aminobezoic acid, 8.2; menadione,.8; retinyl acetate, 0.73; cholecalficerol, 0.003; cyanocobalamin, 0.003. methionine lysine, hematocrit hemoglobin.. ACSBM. ACSBM50 ACSBM60 glucose. Glucose (Gordon, 968),. ACSBM,. Table 6. ACSBM. 9.5% ACSBM 85.9% (82.5%). Table 6. Apparent digestibility coefficient (%, ADC) of dry matter and protein in the test ingredient determined by fecal collection method (Guelph system) for olive flounder Paralichthys olivaceus Test ingredient ADC of test ingredients Acid-concentrated soybean meal., 20%,.. 사 사 (, RP-204-AQ-26) CJ. CJ. References Fish meal Soybean meal Dry matter (%) 57.7±8.0 a 82.0±4.6 b 58.3±7.6 a Protein (%) 85.9±.3 9.5±7.7 82.5±7.8 Mean values of triplicate groups, values are presented as mean ± SD. Values in the same row having different superscript letters are significantly different (P< 0.05). Ao T. 20. Using exogenous enzymes to increase the nutritional value of soybean meal in poultry diet. In: El-Shemy H, Ed. Soybean and Nutrition. InTech, New York, USA, 20-24. AOAC (Association of Official Analytical Chemists), 995. Official Methods of Analysis. 6th edn. Association of Official Analytical Chemists, Arlington, Virginia, USA. Aragão C, Conceicção LEC, Dias J, Marques AC, Gomes E and Dinis MT. 2003. Soy protein concentrate as a protein source for Senegalese sole (Solea senegalensis Kaup 858) diets: effects on growth and amino acid metabolism of postlarvae. Aquac Res 34, 443-452. http://dx.doi.org/0./j.365-209.2003.0097.x. Berge GM, Grisdale-Helland B and Helland SJ. 999. Soy protein concentrate in diets for Atlantic halibut (Hippoglossus hippoglossus). Aquaculture 78, 39-48. http://dx.doi. org/0.06/s0044-8486(99)0027-.

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