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1 한수지 49(3), , 206 Original Article Korean J Fish Aquat Sci 49(3), ,206 항산화기능사료첨가제가조피볼락 (Sebastes schlegelii) 의성장, 체조성, 혈액성상및세균공격성에미치는영향 윤아영 김희성 서영완 조성환 * 배준영 한국해양대학교해양생명과학부, CJ 제일제당 Effects of Dietary Antioxidant Supplementation on the Growth, Serum Chemistry, Body Composition and Challenge Test Results of Juvenile Rockfish Sebastes schlegelii Ahyeong Yun, Hee Sung Kim, Youngwan Seo and Sung Hwoan Cho* and Jun Young Bae Division of Marine BioScience, College of Ocean Science and Technology, Korea Maritime University, Busan 492, Korea Food ingredients New Business Team, CJ CheilJedang Corporation, Seoul 04560, Korea The effects of dietary antioxidant [saltwort (SW), leek (LK), and dandelion (DD)] supplementation on the growth, body composition, serum chemistry, and challenge test results of juvenile rockfish, Sebastes schlegelii, were determined. In an experiment, 320 fish were randomly distributed into eight, 50-L flow-through tanks (40 fish per tank). Four experimental diets were prepared: the control diet (Con) with synthetic antioxidant: and diets with SW, LK, or DD from natural sources. Each diet was randomly assigned to duplicate tanks of fish, which were hand-fed to satiation twice daily for 8 weeks. After the 8-week feeding trial, 20 fish from each tank were artificially infected with Streptococcus iniae and monitored for 6 days. The dietary additives did not affect survival, weight gain, specific growth rate, feed efficiency (FE), protein efficiency ratio (PER), or plasma chemistry. However, the cumulative mortality of fish fed the SW, LK and DD diets was lower than that of fish fed the control diet beginning 4 days after infection. In conclusion, dietary supplementation with SW, LK, and DD did not affect the growth, FE, PER, or plasma chemistry of rockfish. However, dietary inclusion of SW, LK, and DD lowered the mortality of rockfish following infection with S. iniae. Key words: Rockfish (Sebastes schlegelii), Additive, Antioxidant, Challenge test, Streptococcus iniae 서론 (Sebastes schlegelii) (Paralichthys olivaceus) 205 8,774 (45,759 ) (KOSIS, 206). (Kim et al., 200; Wang et al., 2003; Cho et al., 205), (Lee et al., 2000; Lee et al., 203), (Lee et al., 996; Jeon et al., 204) (Kim et al., 999).,., (Tang et al., 200). Korean J Fish Aquat Sci 49(3) , June 206 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial Licens ( which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. Received 27 May 206; Accepted 3 June 206 *Corresponding author: Tel: Fax: address: chosunh@kmou.ac.kr Copyright 206 The Korean Society of Fisheries and Aquatic Science 323 pissn:0374-8, eissn:

2 324 윤아영ㆍ김희성ㆍ서영완ㆍ조성환ㆍ배준영 ( ),. (herb), (Turan and Akyurt, 2005; Turan et al., 2007; Galina et al., 2009; Soosean et al., 200; Dada and Oviawe, 20; Deng et al., 20; Cho and Lee, 202; Kim et al., 203).,. [Saltwort (SW), Salicornia herbacea L.] sterols (Lee et al., 2004), polysaccharides (Lee et al., 2006), flavonoid glycosides (Park and Kim, 2004; Kong et al., 2008) chlorogenic acid (Chung et al., 2005), flavonoid isorhamnetin 3-O-ß-D-glucopyranoside, quercetin 3-O-ß-D-glucopyranoside tungmadic acid (3-caffeoyl-4-dihydrocaffeoyl quinic acid) (Hwang et al., 2009; Kong et al., 2009), (Rhee et al., 2009; Shin et al., 2002; Kim 2007; Song et al., 2007; Im et al., 2003; Jo et al., 2002). [Leek (LK), Allium tuberosum Rottler] flavonoid ß-carotene phenol ß-sitosterol, quercetin, phenolic acid, kaemperol (Wenzel et al., 2000; Jung et al., 2003) (Ahn et al., 200; Jung et al., 2003). [Dandelion (DD), Taraxacum mongolicum H.] flavonoid (Heo and Wang, 2008), (Takasaki et al., 999), (Yoon, 2008) (Han et al., 2005). ethoxyquin (, ),,. 실험어및사육조건 재료및방법 2, 2 ( : 52%, 2%) 2-4%. [ (Mean SE: g)] L ( : 45 L)..4 L/min, aeration. Table. Ingredients (%, DM basis) and nutrient composition of the experimental diets Ingredients (%) Experimental diets Con SW LK DD Fish meal Defatted soybean meal Casein Wheat flour Ethoxyquin 4 0. Additives Squid liver oil Soybean oil Vitamin premix Mineral premix Choline Nutrients (%) Dry matter Crude protein Crude lipid Ash SW (Saltwort), LK (Leek), and DD (Dandelion) were purchased from Tojongherb Co Ltd. (Dongdaemun-gu, Seoul, Korea). 2 Fish meal and was purchased from Abank Co Ltd. (Seocho-gu, Seoul, Korea). 3 Defatted soybean meal was supplied by CJ CheilJedang Corp. (Jung-gu, Seoul, Korea). 4 Ethoxyquin was supplied from Chunhajeil feed Co Ltd. (Daedeok-gu, Daejeon, Korea). 5 Vitamin premix contained the following amount which were diluted in cellulose (g/kg mix): 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-aminobenzoic acid, 8.2; menadione,.8; retinyl acetate, 0.73; cholecalciferol, 0.003; cyanocobalamin, Mineral premix contained the following ingredients (g/kg mix): 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; KI, 0.5; Na 2 Se 2 O 3, 0.0; MnSO 4 H 2 O, 2.0; CoCl 2 6H 2 O,.0.

3 항산화기능사료첨가제가조피볼락에미치는영향 325 실험사료의준비 Table.,,.. 0.% ethoxyquin (, ) 0.% (Con), % SW, LK, DD (,, ) %. 3: (09:00, 7:00). 일반성분분석 8 5,,, AOAC (990). Kjeldahl method (Auto Kjeldahl System, Buchi B-324/435/42, Switzerland) ether-extraction method, 550 C 4, 05 C dry oven 24. 채혈및혈액성상분석 ppm,. Automatic Chemistry System (HITACHI 780/ , Hitachi, Japan) (total protein), glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT) triglyceride. 세균공격성실험 (challenge test) CFU/L Streptococcus iniae (FP5024) 0. ml. 6, 2.,. 통계분석 One-way ANOVA Duncan s multiple range test (Duncan 955) SPSS program version 9.0 (SPSS Michigan Avenue, Chicago, IL, USA). 결과및고찰 8 (%), (g/ fish) (SGR) Table % (P>0.05) g, %/day, (P>0.05). (Choi et al., 2004; Kim et al., 2005; Cho and Lee, 202; Cho et al., 203). 8 (g/fish), (feed efficiency, FE), (protein efficiency ratio, PER) (protein retention, PR) Table 3. (LK) Table 2. Survival (%), weight gain (g/fish) and specific growth rate (SGR) of rockfish Sebastes schlegelii fed the experimental diets containing various sources of antioxidants for 8 weeks Experimental diets Initial weight (g/individual) Final weight (g/individual) Survival (%) Weight gain (g/individual) SGR (%/day) Con 4.3± ± ±2.50 a.2±0.43 a 2.30±0.056 a SW 4.3± ± ±0.00 a.3±0.02 a 2.3±0.003 a LK 4.3± ± ±0.00 a 2.4±0.07 a 2.43±0.0 a DD 4.3± ± ±0.00 a.6±0.02 a 2.34±0.002 a Values (means of duplicate±se) in the same column sharing a common superscript are not significantly different (P>0.05). SGR (%/day) = (Ln final weight of fish - Ln initial weight of fish) 00/days of feeding trial.

4 326 윤아영ㆍ김희성ㆍ서영완ㆍ조성환ㆍ배준영 Table 3. Feed consumption (g/fish), feed efficiency (FE, %), protein efficiency ratio (PER) and protein retention (PR) of rockfish Sebastes schlegelii fed experimental diets containing various sources of antioxidants for 8 weeks Experimental diets Feed consumption (g/fish) FE (%) PER 2 PR 3 Con 0.9±0.32 b 03±. a 2.0±0.09 a 45.4±0.28 a SW.2±0.05 b 00±0.6 a.97±0.03 a 45.2±0.29 a LK 2.3±0.0 a 0±0.5 a.97±0.00 a 42.0±0.35 b DD.6±0.05 ab 00±0.6 a.96±0.0 a 44.0±0.54 a Values (means of duplicate±se) in the same column sharing a common superscript are not significantly different (P>0.05). Feed efficiency (FE, %) = Weight gain of fish 00/feed consumed. 2 Protein efficiency ratio (PER) = Weight gain of fish/protein consumed. 3 Protein retention (PR) = Protein gain 00/protein consumed. (Con) (SW) (P<0.05), (DD) (P>0.05). (00-03%) ( ) (P>0.05). LK (P<0.05). (Cho and Lee, 202), (Choi et al., 2004), (Kim et al., 2005) (Cho et al., 203). 0.5% Chlorella (Bai et al., 200). 8 Table 4. LK DD SW (P<0.05). SW LK DD (P<0.05) (P>0.05). SW (P<0.05), (P>0.05). Chlorella (Bai et al., 200).,, (Seo et al., 2009).,,. Galina et al. (2009) (immunostimulant) (Astragalus membranaceus), (Scutelaria baicalensis), (Lonicera japonica), Ganoderma lucidium holy (Ocimum sanctum), amla (Phyllanthus emblica), neem (Phyllanthus emblica) Solanum trilobatum. 8 total protein, total cholesterol, glutamic pyruvic transaminase (GPT), glutamic oxaloacetic transaminase (GOT) triglyceride Table 5. Table 4. Chemical composition (%) of rockfish Sebastes schlegelii fed experimental diets containing various sources of antioxidants for 8 weeks Experimental diets Moisture Crude protein Crude lipid Ash Con 62.9±0.47 b 2.±0.0 ab 6.2±0.07 b 6.0±0.04 a SW 62.±0.04 b 2.3±0.00 a 6.5±0.04 a 6.2±0.08 a LK 64.8±0.58 a 20.3±0.08 c 6.3±0.03 b 6.0±0.05 a DD 66.±0.3 a 2.0±0. b 6.3±0.00 b 6.±0.04 a Values (means of duplicate±se) in the same column sharing a common superscript are not significantly different (P>0.05). Table 5. Serum chemistry of rockfish Sebastes schlegelii fed experimental diets containing various sources of antioxidants for 8 weeks Experimental diets Total protein (g/dl) Total cholesterol (mg/dl) GPT (IU/L) GOT (IU/L) Triglyceride (mg/dl) Con 4.2±0.9 a 92.0±3.00 a 2.0±.00 a 66.5±8.50 a 265.5±38.50 a SW 4.0±0. a 88.5±8.50 a 2.0±0.00 a 45.0±6.00 a 32.0±26.00 a LK 4.4±0.04 a 96.5±6.94 a 2.5±0.4 a 42.5±6.2 a 307.5± 2.04 a DD 4.5±0.8 a 208.5±7.50 a 3.0±0.00 a 62.5±5.50 a 303.0± 9.00 a Values (means of duplicate±se) in the same column sharing a common superscript are not significantly different (P>0.05). GPT, glutamic pyruvic transaminase; GOT, glutamic oxaloacetic transaminase.

5 항산화기능사료첨가제가조피볼락에미치는영향 327 (P>0.05).. (Jeon et al., 203). 2% glucose (Bai et al., 200) S. iniae 6 Fig.. LK 36, 4 6 SW, LK DD. 4 6 SW LK DD (P<0.05).. 0.5% Vibrio alginolyticus 0.% (Kim et al., 999) % Edwardsiella tarda 60 (Cho and Lee, 202), 0.5% E. tarda (Choi et al., 2004). (Scutellaria baicalensis) E. tarda V. anguillarum S. iniae (Cho et al., 203; Kim et al., 203). glucan E. tarda Cumulative mortality (%) Elapsed time (day) Con SW LK DD Fig.. Cumulative mortality of rockfish Sebastes schlegelii fed the experimental diets containing various sources of antioxidants for 8 weeks, and then infected by Streptococcus iniae (means of duplicates ± SE). (Won et al., 2004).,., S. iniae. 사사 204 ( ) ( ), 2002 ( ) (No. KRF- 202RAA ),. References Ahn JM, Lee SH and Song YS Biological functions in leek. Food Indus Nutr 6, Association of Official Analytical Chemists (AOAC) Official Methods of Analysis (5 th edn). Association of Official Analytical Chemists, Arlington, VA, USA. Bai SC, Koo J, Kim K and Kim S Effects of Chlorella powder as a feed additive on growth performance in juvenile Korean rockfish, Sebastes schlegelii (Hilgendorf). Aquacult Res 32, Cho SH and Lee S Onion powder in the diet of the olive flounder, Paralichthys olivaceus: Effects on the growth, body composition, and lysozyme activity. J World Aquacult Soc 43, x. Cho SH, Jeon GH, Kim HS, Kim DS and Kim C Effects of dietary Scutellaria baicalensis extract on growth, feed utilization and challenge test of olive flounder (Paralichthys olivaceus). Asian-Aust J Anim Sci 26, org/0.573/ajas Cho SH, Kim HS, Myung SH, Jung W, Choi J and Lee S Optimum dietary protein and lipid levels for juvenile rockfish (Sebastes schlegelii, Hilgendorf 880). Aquacult Res 46, Choi S, Ko S, Park G, Lim S, Yu G, Lee J and Bai SC Utilization of song-gang stone as dietary additive in juvenile olive flounder, Paralichthys olivaceus. J Aquaculture 7, Chung YC, Chun HK, Yang JY, Kim JY, Han EH, Kho YH and Jeong HG Tungtungmadic acid, a novel antioxidant,

6 328 윤아영ㆍ김희성ㆍ서영완ㆍ조성환ㆍ배준영 from Salicornia herbacea. Arch Pharm Res 28, Duncan DB Multiple range and multiple F tests. Biometrics, -42. Dada AA, Oviawe NE. 20. The use of bitter kola Garcinia kola dry seed powder as a natural growth-promoting agent for African sharptooth catfish Clarias gariepinus fingerlings. Afr J Aquat Sci 36, Deng J, An Q, Bi B, Wang Q, Kong BL, Tao L and Zhang X. 20. Effect of ethanolic extract of propolis on growth performance and plasma biochemical parameters of rainbow trout (Oncorhynchus mykiss). Fish Physiol Biochem 37, Galina J, Yin G, Ardo L and Jeney Z The use of immunostimulating herbs in fish. An overview of research. Fish Physiol Biochem 35, Han SH, Hwang JK, Park SN, Lee KH, Kang IK, Kim KS and Kim KH Potential effect of solvent fractions of Taraxacum mongolicum H. on protection of gastric mucusa. Korean J Food Sci Technol 37, Heo S, Wang M Antioxidant activity and cytotoxicity effect of extracts from Taraxacum mongolicum H. Kor J Pharmacogn 39, Hwang YP, Yun HJ, Chun HK, Chung YC, Kim HK, Jeong MH, Yoon TR and Jeong HG Protective mechanisms of 4-caffeoyl, 4-dihydrocaffeoyl quinic acid from Salicornia herbacea against tert-butylhydroperoxide-induced oxidative damage. Chemico-Biological Interactions 8, Im SA, Kim GW and Lee CK Immunomodulatory activity of Salicornia herbacea L. components. Korean J Nutr 40, 5-3. Jeon GH, Cho SH, Lee SM, Nam TJ and Kim DS Effects of the dietary inclusion of Porphyra and sea tangle Laminaria japonica on the growth, feed utilization, body composition, and plasma chemistry of juvenile Korean rockfish Sebastes schlegelei. Korean J Fish Aquat Sci 46, Jeon GH, Kim HS, Myung SH and Cho SH The effect of the dietary substitution of fishmeal with tuna by-product meal on growth, body composition, plasma chemistry and amino acid profiles of juvenile Korean rockfish (Sebastes schlegelii). Aquacult Nutr 20, org/0./anu.253. Jo YC, Ahn JH, Chon SM, Lee KS, Bae TJ and Kang DS Studies on pharmacological effects of glasswort (Salicornia herbacea L.). Korean J Med Crop Sci 0, Jung H, Noh K, Cho H, Park J, Choi C, Kwon T and Song Y Effect of buchu (Allium tuberosum) on lipid peroxidation and antioxidative defense system in streptozotocininduced diabetic rats. Korean J Life Sci 3, Kim MW Effects of Salicornia herbacea L. supplementation on blood glucose and lipid metabolites in streptozotocin-induced diabetic rats. Korean J Nutr 40, 5-3. Kim KH, Hwang YJ and Bai SC Resistance to Vibrio alginolyticus in juvenile rockfish (Sebastes schlegelii) fed diets containing different doses of aloe. Aquaculture 80, Kim K, Wang XJ and Bai SC Reevaluation of the optimum dietary protein level for the maximum growth of juvenile Korean rockfish, Sebastes schlgeli (Hilgendorf). Aquacult Res 32 (Suppl. ), org/0.046/j x x. Kim C, Kang B, Jee H, Choi A, Kim C, Cho Y, Hahn H and Nam KD Effect of chitosan-based feed additive on the growth and quality of cultured Japanese flounder, Paralichthys olivaceus. J Chitin Chitosan 0, Kim KT, Jeon GH, Cho SH, Lim SG, Kwon M and Yoo J Effects of dietary inclusion of various concentrations of Scutellaria baicalensis Georgi extract on growth, body composition, serum chemistry and challenge test of far eastern catfish (Silurus asotus). Aquacult Res 44, dx.doi.org/0./j x. KOSIS Korean Statistical Information Service. Korea. Kong C, Kim YA, Kim M, Park J, Kim J, Kim S, Lee B, Nam TJ and Seo Y Flavonoid glycosides isolated from Salicornia herbacea inhibit matrix metalloproteinase in HT080 cells. Toxicol in Vitro 22, org/0.06/j.tiv Kong C, Kim JA, Qian ZJ, Kim YA, Lee JI, Kim SK, Nam TJ and Seo YW Protective effect of isorhamnetin 3-Oß-D-glucopyranoside from Salicornia herbacea against oxidation-induced cell damage. Food Chem Taxicol 47, Lee S, Yoo J and Lee JY The use of soybean meal, corn gluten meal, meat meal, meat and bone meal, or blood meal as a dietary protein source replacing fish meal in Korean rockfish (Sebastes schlegelii). Korean J Ani Nut Feed 20, Lee S, Hwang U and Cho SH Effects of feeding frequency and dietary moisture content on growth, body composition and gastric evacuation of juvenile Korean rockfish (Sebastes schlegelii). Aquaculture 87, org/0.06/s (00) Lee YS, Lee HS, Shin KH, Kim BK and Lee SH Constituents of the halophyte Salicornia herbacea. Arch Pharm Res 27, /BF Lee KY, Lee MH, Chang IY, Yoon SP, Lim DY and Jeon YJ Macrophage activation by polysaccharide fraction isolated from Salicornia herbacea. J Ethnopharm 03, Lee J, Lee B, Kim K, Han H, Park G, Lee J, Yun H and Bai SC Optimal feeding frequency for juvenile Korean rock-

7 항산화기능사료첨가제가조피볼락에미치는영향 329 fish Sebastes schlegelii fed commercial diet at two different water temperatures. Korean J Fish Aquati Sci 46, Park S and Kim K Isolation and identification of antioxidant flavonoids from Salicornia herbacea L. J Korean Soc Appl Biol Chem 47, Rhee KS, Park HJ and Cho JY Salicornia herbacea: botanical, chemical and pharmacological review of halophyte marsh plant. J Med Plants Res 3, Seo J, Kim K and Lee S Effects of supplemental herb medicines in the diets on growth, feed utilization and body composition of juvenile and grower rockfish Sebastes schlegelii. J Aquaculture 22, 2-7. Shin KS, Boo HO, Jeon MW and Ko JY Chemical components of native plant, Salicornia herbacea L. Korean J Plant Res 5, Song H, Kim D, Jung Y and Lee M Antioxidant activities of red hamcho (Salicornia herbacea L.) against lipid peroxidation and the formation of radicals. Korean J Food Nutr 20, Soosean C, Marimuthu K, Sudhakaran S and Xavier R Effect of mangosteen (Garcinia mangostana L.) extracts as a feed additive on growth and hematological parameters of African catfish (Clarias gariepinus) fingerlings. Eur Rev Med Pharmacol Sci 4, Takasaki M, Konoshima T, Tokuda H, Masuda K, Arai Y, Shiojima K and Ageta H Anti-carcinogenic activity of Taraxacum plant. I. Biol Pharm Bull 22, Tang S, Kerry JP, Sheehan D and Buckley DJ A comparative study of tea catechins and α-tocopherol as antioxidants in cooked beef and chicken meat. Eur Food Res Technol 23, Turan F and Akyurt I Effects of red clover extract on growth performance and body composition of African catfish Clarias gariepinus (Bruchell, 822). Fish Sci 7, Turan F, Güurek M, Yaglioglu D Dietary red clover (Trifolium pratense) on growth performance of common carp (Cyprinus carpio). J Ani Vet Adv 6, Wang X, Kim K, Park G, Choi S, Jun H and Bai SC Evaluation of L-ascorbyl-2-glucose as the source of vitamin C for juvenile Korean rockfish Sebastes schlegelii (Hilgendorf). Aquacult Res 34, org/0.046/j x. Wenzel U, Kuntz S, Brendel MD and Daniel H Dietary flavone is a potent apoptosis inducer in human colon carcinoma cells. Cancer Res 60, Won K, Kim S and Park S The effects of β-,3/,6-linked glucan in the diet on immune responses of olive flounder, Paralichthys olivaceus by oral administration. J Fish Pathol 7, Yoon T Effect of water extracts from foot of Taraxacum officinale in innate and adaptive immune responses in mice. Korean J Food Nutr 2,

한수지 47(6), , 2014 Original Article Kor J Fish Aquat Sci 47(6), ,2014 배합사료및습사료공급에따른넙치 (Paralichthys olivaceus) 의성장및어체성분비교 김강웅 이진혁 배기민 김경덕

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