한수지 52(2), , 2019 Original Article Korean J Fish Aquat Sci 52(2), ,2019 동 식물성혼합물의사료내어분대체가강도다리 (Platichthys stellatus) 의성장및혈액성상에미치는영향 신승준

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한수지 52(2), 34-40, 209 Original Article Korean J Fish Aquat Sci 52(2),34-40,209 동 식물성혼합물의사료내어분대체가강도다리 (Platichthys stellatus) 의성장및혈액성상에미치는영향 신승준 이태규 이동훈 임성률 양일창 김성삼 2 최정우 김진수 김정대 * 강원대학교동물생명과학대학, 경기도해양수산자원연구소, 2 CJ 제일제당 Effect of Dietary Fish Meal Replacement by a Blend of Plant and Animal Ingredients on the Growth and Blood Chemistry of Starry Flounder Platichthys stellatus Seung-Jun Shin, Tae-Kyu Lee, Dong-Hoon Lee, Seong-Ryul Lim, Il-Chang Yang, Sung-Sam Kim 2, Jung-Woo Choi, Jin-Soo Kim and Jeong-Dae Kim* College of Animal Life Sciences, Kangwon National University, Chuncheon 2434, Korea Gyenoggi Province Maritime and Fisheries Research Institute, Yangpyeong 253, Korea 2 CJ Cheiljedang, Feed & Livestock R&D, Suwon 6495, Korea A feeding trial was conducted to investigate the effects of dietary fish meal replacement by a blend of lysine cell mass, corn protein concentrate and poultry by-product meal on the growth and blood chemistry of the starry flounder Platichthys stellatus. The fish meal replacer (FMR) was prepared to have the same level of protein as fish meal (FM). With a commercial diet as a positive control, five experimental diets (basal, FM42, FM32, FM22 and FM2) were formulated to contain 52% protein and 0% lipid. The dietary FM levels decreased from 52% (basal) to 42, 32, 22 and 2% with concomitant increase in the FMR to 0, 20, 30, 40 and 50%, respectively. Juvenile starry flounder with an average body weight of 77.3 g were randomly distributed in each (30 fish/tank) of 8 plastic tanks (39 99 54 cm). After a 45-day feeding trial, the survival rate ranged from 95.6% (FM22) to 00% (control and FM42), while the weight gain of the fish groups varied from 49.7 to 58.4 g. The results clearly revealed that starry flounder can grow well on a diet containing low FM (2%) with a high level of FMR (50%) without any adverse effects. Key words: Platichthys stellatus, Fish meal, Fish meal replacer (FMR), Survival rate, Weight gain 서론 (Platichthys stellatus),., 5. 60% (Gatlin et al., 2007; Tacon et al., 20).,. (Olsen and Hasan, 202). (Lim et al., 2008). *Corresponding author: Tel: +82. 33. 250. 8634 Fax: +82. 33. 259. 5572 E-mail address: menzang@gmail.com 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. https://doi.org/0.5657/kfas.209.034 Korean J Fish Aquat Sci 52(2), 34-40, April 209 Received 4 January 209; Revised 2 February 209; Accepted 9 March 209 저자직위 : 신승준 ( 연구원 ), 이태규 ( 연구원 ), 이동훈 ( 연구사 ), 임성률 ( 연구사 ), 양일창 ( 연구원 ), 김성삼 ( 연구원 ), 최정우 ( 교수 ), 김진수 ( 교수 ), 김정대 ( 교수 ) Copyright 209 The Korean Society of Fisheries and Aquatic Science 34 pissn:0374-8, eissn:2287-885

강도다리사료내어분대체제의첨가효과 35, (Lim et al., 2008; Krogdahl et al., 200; NRC, 20). (Thiesses et al., 2003; Barrows et al., 2007; Burr et al., 202). (Storebakken et al., 2000; Refstie et al., 2006). (Lim et al., 2008; Gaylord and Barrows, 2009). (Burr et al., 202). (EP, extrude pellet). (Lee et al., 2006), (Lee et al., 2003), (Lee et al., 2004) (Park et al., 206)., (lysine cell mass). 재료및방법 실험어및사육조건 2,000., 77.3 4.7 g (mean+sd) 550 L (39 99 54 cm) 8 30 3. 7.7 0.2 mg/l,.6.. % ( 4 ). 2, 45. 실험사료및실험설계 Table Table 2. (fish meal), (soybean meal). (fish oil) (wheat flour). -2 [mono-dicalcium phosphate, Ca(H 2 PO 4 ) 2 H 2 O+CaHPO 4 2H 2 O]. ( Table. Ingredient and chemical composition of the experimental diets Ingredient Fish meal, sardine Basal FM42 FM32 FM22 FM2 Control 52.00 42.00 32.00 22.00 2.00Closed FMR 0.00 20.00 30.00 40.00 50.00 - Soybean meal 2.00 2.00 2.00 2.00 2.00 - SPC 2 3.00 3.00 3.00 3.00 3.00 - Wheat flour 4.65 3.30 2.25.05 0.00 - Lecithin 0.50 0.50 0.50 0.50 0.50 - Fish oil 3.50 3.90 4.00 4.20 4.50 - Amino acids 3.40 2.05 2.70 3.40 4.05 - MDCP 4 0.40 0.60 0.80.00.00 - Mineral mixture 0.80 0.90.00.0.20 - Vitamin. E 0.30 0.30 0.30 0.30 0.30 - Choline 0.30 0.30 0.30 0.30 0.30 - Vitamin. C 0.30 0.30 0.30 0.30 0.30 - Vitamin mixture 0.80 0.80 0.80 0.80 0.80 - Mold inhibitor 0.05 0.05 0.05 0.05 0.05 - Total 00.00 00.00 00.00 00.00 00.00 00.00 Proximate composition (g/00g, as-fed) Moisture 9.5 9.24 9.38 9.35 9.38 9.22 Crude protein 5.60 5.94 52.08 52.06 52.5 5.53 Crude fat 9.35 9.77 9.9 9.89 0.09 0.56 Crude fiber.6.25.5.54.57 0.86 Crude ash 8.62 8.36 8.06 7.65 5.24 9.60 Pepsin Digestibility 89. 86.58 9.77 9.68 88.3 89.29 Fish meal replacer, a blend of lysine cell-mass (Bioprotam, Daesang Corporation, Korea), corn protein concentrate (Empyreal 75, Cargill, Inc., USA) and poultry-by product meal (Harim Corp., Korea), which were mixed at the ratio of :.5:2.5 to contain the same level of protein as fish meal. 2 Soy Protein Concentrate, Selecta, Brazil. 3 Contain lysine, methionine, tryptophan, betaine and taurine. 4 Mono-, di-calcium phosphate. FM, Fish meal.

36 신승준ㆍ이태규ㆍ이동훈ㆍ임성률ㆍ양일창ㆍ김성삼ㆍ최정우ㆍ김진수ㆍ김정대, ) 4 6. 어체측정, 24 (AQUI-S, New Zealand Ltd., Lower Hutt, NZ) (weight gain, %), (survival rate, %), (specific growth rate, %), (daily feed intake, %/average weight/d), (feed conversion rate) (protein efficiency ratio). 일반성분및아미노산분석 5, AOAC (995) (25, 3 ), kjeldahl (N 6.25),. 2 soxtec system 046 (Tacator AB, Sweden) soxhlet. 6 N HCl ninhydrin Table 2. Amino acid profile of the experimental diets Amino acid Basal FM42 FM32 FM22 FM2 Control Aspartic acid 4.6 4.46 4.48 4.27 4.26 4.38 Threonine 2.5 2.08 2.0 2.0 2.0 2.09 Serine 2.8 2.2 2.5 2.06 2.07 2.3 Glutamic acid 7.29 7.3 7.23 6.97 7.08 7.0 Proline 2.72 2.70 2.75 2.6 2.68 2.58 Glycine 3.66 3.58 3.53 3.23 3.8 3.08 Alanine 3.3 3.08 3.2 3.07 3.9 2.83 Valine 2.44 2.35 2.39 2.30 2.35 2.44 Isoleucine 2.09 2.02 2.04.95.97 2.06 Leucine 3.7 3.64 3.65 3.56 3.65 3.65 Tyrosine.54.45.42.28.43.48 Phenylalanine 2.08.99.99.93 2.0 2.06 Histidine.3.07.09.02.0.2 Lysine 3.66 3.5 3.56 3.40 3.40 3.82 Arginine 3.7 3.0 3.3 2.98 2.98 2.98 Tryptophan 0.55 0.54 0.52 0.5 0.50 0.6 Taurine 0.30 0.33 0.36 0.40 0.43 0.26 Cystine 0.56 0.55 0.55 0.52 0.53 0.65 Methionine.6...05.07.2 Total 47.28 45.94 46.38 44.2 44.87 45.66 FM, Fish meal. amino acid Analyzer S433 (SYKAM, German). 혈액성분분석 hematocrit (PCV, %) hemoglobin (Hb, g/ dl), 4, 2,000 rpm, 0 (Centrifuge 545R, Eppendorf Ltd., Germany) (plasma), aspartate aminotransferase (AST, U/L), alanine aminotransferase (ALT, U/L), albumin (ALB, g/dl), glucose (GLU, mg/dl), total cholesterol (TCHO, mg/dl), triglyceride (TG, mg/dl) total protein (TP, g/dl). PCV HAEMA- TOKRIT20 (Hettich Ltd., Germany), Hb (Fuji DRI- CHEM slide, Fuji photo film co. Ltd., Japan) (DRI-CHEM 3500 I, Fujifilm Ltd., Japan).. 통계처리 (,,,,,,, ), SPSS Version 0 (SPSS, Michigan Avenue, Chicago, IL, USA) (one-way ANOVA). (homogeneity of variance) Cochran s test (Sokal and Rohlf, 995)., Duncan s multiple range test (Duncan, 955). 결과 45 Table 3. (WG, weight gain, %) (P>0.05) (27.80%). 49.7 g ( ) 58.4 g (FM42, FM32), 95.6% (FM22) 00% (, FM42) (P>0.05). (DFI, daily feed intake).03-.05% (SGR, specific growth rate, %).06% FM32.24% (P>0.05). (FCR, feed conversion ratio) FM42 FM2.50,.76 (P>0.05).

강도다리사료내어분대체제의첨가효과 37 (PER, protein efficiency ratio).00-.7 (P>0.05). Table 4, 5. (PCV, %) FM42 FM32, FM22 (P<0.05) (basal) FM2 (P>0.05)., FM32 FM22 (P<0.05) FM32, FM22 (P>0.05). (Hb, g/dl) (P>0.05)., aspartate aminotransferase (AST) 25.8-3.2 U/L, FM32, FM22 (P<0.05), FM42 FM2 (P>0.05)., FM42 FM22 (P<0.05), FM32, FM22 (P>0.05). alanine aminotransferase (ALT) 80.2-06.0 U/L FM32, FM22 (P<0.05), FM42 FM2 (P>0.05)., FM42 FM22 (P<0.05), FM32, FM22 Table 3. Growth performance of starry flounder Platichthys stellatus fed the experimental diets for 45 days WG Diet In.wt (g/f) 2 Fin.wt (g/f) 3 % 4 g/fish 5 Survival 6 DFI 7 SGR 8 FCR 9 PER 0 Basal 76.89±3.3 ns 23.7±5.0 ns 30.68±.34 ns 54.28±2.70 ns 96.67±3.33 ns.05±0.0 ns.6±0.05 ns.62±0.08 ns.09±0.95 ns FM42 78.44±3.49 236.89±6.73 32.73±2.5 58.44±4.27 00.00±0.00.03±0.0.23±0.08.50±0.09.7±0.3 FM32 77.78±3.32 236.7±4.38 32.9±2.86 58.39±4.43 96.67±.93.04±0.0.24±0.09.52±0.2.5±0.6 FM22 77.±2.9 235.9±4.48 32.77±0.98 58.08±2.39 95.56±2.94.05±0.02.23±0.04.5±0.06.5±0.07 FM2 75.±3.20 23.93±5.53 32.42±0.79 56.82±2.37 98.89±..03±0.00.22±0.03.50±0.04.5±0.05 Control 78.22±3.93 227.89±7.95 27.80±.6 49.67±4.02 00.00±0.00.05±0.00.06±0.06.76±0.08.00±0.08 Values (means±se of the triplicate replications) in the same column with the same superscript are not significantly different (P>0.05); ns, nonsignificant. 2 Initial weight, g/fish. 3 Final weight, g/fish. 4 Weight gain (%)=(final weight-initial weight)/final weight 00. 5 Weight gain (g/fish)=final weight-initial weight. 6 Survival rate (%)=final fish number/initial fish number 00. 7 Daily Feed intake (%/av. bw/d)=feed intake (as-is)/[(initial body weight+final body weight)/2]/45 00. 8 Specific growth rate (%/d)=[ln(final wt)-ln(initial weight)]/experimental days 00. 9 Feed conversion ratio=dry feed intake/wet weight gain. 0 Protein efficiency ratio=weight gain/protein intake. FM, Fish meal. Table 4. Haematological characteristics of juvenile starry flounder Platichthys stellatus fed the experimental diets for 45 days Parameters Basal FM42 FM32 FM22 FM2 Control PCV (%) 2 27.2±.7 ab 29.0±2.4 a 20.7±.3 d 22.5±0.5 cd 25.7±.0 abc 24.0±.3 bcd Hb (g/dl) 3 5.6±0.3 5.7±0.4 4.9±0. 5.0±0. 5.2±0. 5.2±0. ns Values (means±se of the triplicate replications) in the same row with the same superscript are not significantly different (P>0.05). 2 Hematocrit. 3 Hemoglobin. FM, Fish meal. Table 5. Blood plasma parameters of juvenile starry flounder Platichthys stellatus fed the experimental diets for 45 days Parameters Basal FM42 FM32 FM22 FM2 Control AST 2 3.2±0.7 a 29.7±.0 ab 27.0±0.7 bc 25.8±.2 c 30.3±0.7 a 27.3±.3 bc ALT 3 06.0±3.5 a 99.2±3.8 ab 94.3±2.9 bc 80.2±3.7 d 98.7±4.0 ab 85.0±3.7 cd ALB 4 0.9±0. a 0.8±0. ab 0.7±0. b 0.7±0. b 0.8±0. ab 0.8±0. ab GLU 5 85.3±2.5 a 79.0±2.8 ab 60.5±3.6 c 60.2±3.9 c 66.8±4. c 69.2±4.4 bc TCHO 6 232.0±8.7 a 203.7±4.2 b 82.8±5.7 b 89.2±6.6 b 200.5±4.6 b 202.3±2.5 b TG 7 30.3±4.9 a 5.0±4.8 b 07.8±2.7 c.8±4.0 ab.3±2. b 08.5±.2 b TP 8 4.2±0. a 3.8±0. bc 3.6±0. c 3.7±0. c 4.±0. ab 3.8±0. bc Values (means±se of the triplicate replications) in the same row with the same superscript are not significantly different (P>0.05); ns, nonsignificant. 2 Aspartate aminotransferase, U/L. 3 Alanine aminotransferase, U/L. 4 Albumin, g/dl. 5 Glucose, mg/dl. 6 Total cholesterol, mg/dl. 7 Triglyceride, mg/dl. 8 Total protein, g/dl. FM, Fish meal.

38 신승준ㆍ이태규ㆍ이동훈ㆍ임성률ㆍ양일창ㆍ김성삼ㆍ최정우ㆍ김진수ㆍ김정대 (P>0.05). (ALB, albumin) 0.7-0.9 g/dl FM32, FM22 (P<0.05), FM42, FM2 (P>0.05)., (P>0.05). (GLU, glucose) 60.2-85.3 mg/dl FM32, FM22, FM2 (P<0.05), FM42 (P>0.05)., FM42 FM32, FM22 FM2 (P<0.05), FM32, FM22, FM2 (P>0.05). (TCHO, total cholesterol) 82.8-232.0 mg/dl (P<0.05), (P>0.05). (TG, triglyceride) 07.8-30.3 mg/dl FM42, FM32, FM2 (P<0.05), FM22 (P>0.05). (TP, total protein) 3.6-4.2 g/ dl FM42, FM32, FM22 (P<0.05), FM2 (P>0.05). Table 6, (P>0.05). 고찰 40-60%, (Karapanagiotidis, 204). (CPC, corn protein concentrate) wet-milling (endosperm), LCM (lysine cell mass) Corynebacterium, (PBPM, poultry by-product meal),. Burr et al. (202) ( 3.5 g) CPC 27.3%, 38.6%,. Yu et al. (203) (Litopenaeus vannamei) 2% CPC,,. Khalifa et al. (208) 20% 4.5%, 9.3% 0% CPC 5%, 0% 9% 8,.5 g 3. g ( ), 2.5 g (CPC 5% ) 2.4 g (CPC 0% ), CPC 9% 0. g. LCM,. LCM (ammonium sulfate), (crude protein) 68.6%, 0.4% LCM. Wang et al. (2004) (Sebastes schlegeli) LCM (BASF Ltd.) 0%, LCM 70% 0.07%. (SCP, single cell protein) (yeast), (algae), SCP 970. (algae), (yeast). SCP Table 6. Muscle composition of juvenile starry flounder Platichthys stellatus fed the experimental diets for 45 days Composition Diet Basal FM42 FM32 FM22 FM2 Control Moisture 76.26±0.5 76.58±0.65 76.4±0.24 76.2±0.2 75.56±0.7 76.59±0.07 ns Crude fat 3.±0.4 3.00±0.6 2.87±0.08 2.80±0.32 3.05±0.47 2.70±0.25 ns Crude protein 9.59±0.37 9.8±0.5 9.93±0.5 20.09±0.07 20.37±0.26 9.83±0.22 ns Values (means±se of the triplicate replications) in the same row with the same superscript are not significantly different (P>0.05); ns, nonsignificant. FM, Fish meal.

강도다리사료내어분대체제의첨가효과 39 (Francis et al., 200; Refstie et al., 2005)., LCM. (PBPM) 64.8%, 2.99%,,, 0.93, 0.40 0.24% PBPM., ( 5.66 g) 70% (Song et al., 204). LCM, CPC PBPM (FMR) (Table ), (basal) 52% FMR 0%, FM2 2% FMR 50%, 40% FMR 40%. Table 3 Table 6 FMR 0% (basal) 50% (FM2), (control group), FMR. (,, )., Ding et al. (200) 45%, 0% TP, TG TCHO 2, Park et al. (206) AST ALT 20.5 3.5 U/L, ALT (Table 5). Lee et al. (2003).9 g 0 AST 37-87 U/L, ALT 9-97 U/L.., EP. FMR, FMR. 사사 207 ( : 52070239), ( : 2060883) ( : 2060744). References AOAC (Association of Official Analytical Chemists). 995. Official Methods of Analysis. 6th ed. Arlington, VA, U.S.A. Barrows FT, Gaylord TG, Stone DAJ and Smith CE. 2007. Effect of protein source and nutrient density on growth efficiency, histology, and plasma amino acid concentration of rainbow trout (Oncorhynchus mykiss). Aquac Res 38, 747-758. https://doi.org/0./j.365-209.2007.0854.x. Burr GS, Wolters WR, Barrows FT and Hardy RW. 202. Replacing fishmeal with blends of alternative proteins on growth performance of rainbow trout (Oncorhynchus mykiss), and early or late stage juvenile Atlantic salmon (Salmo salar). Aquaculture 334-337, 0-6. https://doi. org/0.06/j.aquaculture.20.2.044. Ding L, Zhang L, Wang J, Ma J, Meng X, Duan P, Sun L and Sun Y. 200. Effect of dietary lipid level on the growth performance, feed utilization, body composition and blood chemistry of juvenile starry flounder (Platichthys stellatus). Aquac Res 4, 470-478. https://doi.org/0./j.365-209.2009.02440.x. Duncan DB. 955. Multiple range and multiple F tests. Biometrics, -42. Francis G, Makkar HP and Becker K. 200. Antinutritional factors present in plant-derived alternate fish feed ingredients and their effects in fish. Aquaculture 99, 97-227. https:// doi.org/0.06/s0044-8486(0)00526-9. Gatlin DM, Barrows FT, Brown P, Dabrowski K, Gaylord TG, Hardy RW, Herman E, Hu G, Krogdahl Å, Nelson R, Overturf K, Rust M, Sealey W, Skonberg D, Souza EJ, Stone D, Wilson R and Wurztel E. 2007. Expanding the utilization of sustainable plant products in aquafeeds: a review. Aquac Res 38, 55-579. https://doi.org/0./j.365-209.2007.0704.x. Gaylord TG and Barrows FT. 2009. Multiple amino acid supplementation to reduce dietary protein in plant-based rainbow trout, Oncorhynchus mykiss, feeds. Aquaculture 287, 80-84. https://doi.org/0.06/j.aquaculture.2008.0.037.

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