58 윤태현ㆍ이초롱ㆍ차지훈ㆍ김주민ㆍ신승준ㆍ이경준ㆍ김정대 5.9% (KOSIS, 207). 25,652 92%, 70,000 (Kim, 207). Choi et al. (2005) 4 g % % (Kim, 204), 2.,. 55%,. 재료

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한수지 5(2), 57-62, 208 Original Article Korean J Fish Aquat Sci 5(2),57-62,208 육성넙치 (Paralichthys olivaceus) 사료내인공급제의첨가가성장률, 사료이용효율, 혈액성상및어체내광물질조성에미치는영향 윤태현 이초롱 차지훈 김주민 신승준 이경준 김정대 * 강원대학교동물생명과학대학, 제주대학교해양생명과학과 Supplemental Effects of Dietary Phosphorus on the Growth, Feed Utilization, Blood Parameters and Mineral Composition of the Bodies of Juvenile Olive Flounders Paralichthys olivaceus Tae-Hyun Yoon, Chorong Lee, Ji-Hoon Cha, Joo-Min Kim, Seung-Jun Shin, Kyeong-Jun Lee and Jeong-Dae Kim* College of Animal Life Sciences, Kangwon National University, Chuncheon 2434, Korea Department of Marine Life Sciences, Jeju National University, Jeju 63243, Korea A feeding trial was conducted to investigate the effects of supplementation with four phosphorus (P) additives [monocalcium phosphate (MCP), dicalcium phosphate (DCP), tricalcium phosphate (TCP) and magnesium hydrogen phosphate (MHP)] on the growth, feed utilization and whole body mineral composition of juvenile olive flounders Paralichthys olivaceus. A basal diet without P supplementation was prepared as a negative control and four supplemental P sources were added at a level of 2%. Triplicate groups of fish (initial mean body weight g) were fed one of the experimental diets to apparent satiation twice a day, at 08:30 and 8:00 for 0 weeks. The final body weights of fish fed the experimental diets ranged from 29.4 g (DCP) to 34.2 g (MCP) and did not differ significantly (P>0.05) among treatments. Similar tendencies were found for all parameters including weight gain (%), specific growth rate (SGR), feed conversion ratio (FCR), protein efficiency ratio (PER), feed intake (FI) and survival rate (SR). The hematocrit (%), hemoglobin (g/dl), serum inorganic P and whole body mineral composition did not differ significantly different (P>0.05) among the treatments. Therefore, dietary P addition is not necessary for juvenile olive flounder fed a fish meal-based diet. Key words: Paralichthys olivaceus, Phosphorus, Growth, Feed conversion ratio, Mineral composition 서론 (P), (Auer et al., 986)., 0.02-0.6 mg/l (Lall, 99). 0.3 0.9% (NRC, 20), (monocalcium phosphate, MCP), (dicalcium phosphate, DCP) (tricalcium phosphate, TCP). (Yoon et al., 204; 205; 206) (total phosphorus) (Yoon et al., 207). (Paralichthys olivaceus) 206 4,636 https://doi.org/0.5657/kfas.208.057 Korean J Fish Aquat Sci 5(2) 57-62, April 208 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 8 March 208; Revised 26 March 208; Accepted 30 March 208 *Corresponding author: Tel: +82. 33. 250. 8634 Fax: +82. 33. 259. 5570 E-mail address: menzang@gmail.com Copyright 208 The Korean Society of Fisheries and Aquatic Science 57 pissn:0374-8, eissn:2287-885

58 윤태현ㆍ이초롱ㆍ차지훈ㆍ김주민ㆍ신승준ㆍ이경준ㆍ김정대 5.9% (KOSIS, 207). 25,652 92%, 70,000 (Kim, 207). Choi et al. (2005) 4 g 6 0.45-0.57%. 50-70% (Kim, 204), 2.,. 55%,. 재료및방법 실험사료의제조. (control) MCP (monocalcium phosphate), DCP (dicalcium phosphate), TCP (tricalcium phosphate) MHP (magnesium hydrogen phosphate, MgHPO 4 ) 2% (Kim et al., 998) 4 5. 50% 0%, 55%,.5%, 3%, 8%, 3%. 3% %.. 6%, 37.%.9%, 2% 2% (Table ). 00 mesh (40 µm). 500 kg (Table ). V-mixer (Hangjin Co., Korea) 0. 5 twin extruder (model ATX-2, Fesco Precision Co., Korea) 2 mm 3 mm. Table extrusion. Feeder speed 6-8 rpm, conditioner temperature 80-90, 00-50 ml, screw speed 250-320 rpm, st barrel 50 ml, 2 nd barrel 3 rd barrel temperature 05-35, steam heater pressure 4-6 kgf/cm², 4 th barrel temperature 80-90., 60-80 6 5-8%. 실험어및사육관리 ( Table. Ingredient and chemical composition of the experimental diets Ingredient (%) Experimental diets Control MCP DCP TCP MHP Fish meal 55.00 55.00 55.00 55.00 55.00 Soybean meal.50.50.50.50.50 Wheat flour 8.00 8.00 8.00 8.00 8.00 Squid liver powder 3.00 3.00 3.00 3.00 3.00 Fish oil 3.00 3.00 3.00 3.00 3.00 Vitamin mix 2 0.0 0.0 0.0 0.0 0.0 Mineral mix 3 0.0 0.0 0.0 0.0 0.0 Corn gluten meal 3.00 3.00 3.00 3.00 3.00 Finnstim 4.00.00.00.00.00 Choline-HCl 5 0.70 0.70 0.70 0.70 0.70 P source - 2.00 2.00 2.00 2.00 Corn grain 4.60 2.60 2.60 2.60 2.60 Total 00.00 00.00 00.00 00.00 00.00 Composition (%, DM) 6 C. protein 5.32 50.34 49.68 49.29 49.78 C. lipid 0.30 9.88 0.08 9.57 0.26 C. ash 2.59 4.33 4.35 5.78 5.55 CHO 27.79 25.45 25.89 25.36 24.4 Ca 3.2 3.69 3.60 3.86 3.79 P 2.3 2.49 2.50 2.54 2.5 MCP, monocalcium phosphate; DCP, dicalcium phosphate; TCP, tricalcium phosphate; MHP, magnesium hydrogen phosphate. 2 Vitamin added to supply the following (per kg diet): vitamin A, 8000 IU; vitamin D 3, 600 IU; vitamin E, 200 IU; vitamin K 3, 30 mg; thiamine HCl, 80 mg; riboflavin, 50 mg; D-Ca pantothenate, 00 mg; biotin, mg; folic acid, 0 mg; vitamin B 2, 80 mg; niacin, 200mg; pyridoxine HCl,40 mg; ascorbic acid, 500 mg; inositol, 400 mg; BHT, 5 mg; BHA, 5 mg. 3 Mineral added to supply the following (per kg diet): ferrous sulfate (20.% Fe), 220 mg; copper sulfate (25.4% Cu), 63.5 mg; zinc sulfate (22.7% Zn), 352 mg; manganous sulfate (32.5% Mn), 00 mg; cobalt chloride (24.8% Co), 20 mg; potassium iodide (76.4% I), 6.5 mg; sodium selenite (45.6% Se), 2.2 mg; sodium fluoride (45.2% F), 8 mg. 4 Betaine S, DANISCO, Finland. 5 Coline Chloride-50, Cofavet special, Korea. 6 Values are means of 2 or 3 determinations.

넙치사료내인공급제첨가효과 59 ). 2 3 FRP. (, g) 5 20 L (3 / ) 35. 3 L/min. 2L:2D, 3-8.5. 2 (08:30 hr 8:00 hr). 4 (0 ), 24. 4 2 mm 6 3 mm, 0. 샘플수집및생화학적분석 0, (weight gain), (specific growth rate), (feed conversion ratio), (protein efficiency ratio) (survival rate)., 5 2-phenoxyethanol (200 ppm). 5 ( 5 ) hematocrit, hemoglobin inorganic P. AOAC (995) (25, 3 ), (550, 6 ), (Kjeltec System 2300, Sweden), Folch et al. (957) Soxhlet (Soxhlet Heater System C-SH6, Korea)..0 g, (START-D microwave digestion system, Milestone, USA). (PFA, Perfluoroalkoxy) 0 ml 6,. 5 80 5 0, 200-250 psi. 0.45 m 25 ml. ICP-OES. ICP (Inductively Coupled Plasma) 000 g/ml (AccuStandard, USA). HNO 3. 0.0-5.0 g/ml, 0.0-.0 g/ml ICP-OES (Perkin Elmer OPTIMA 7300DV, USA). Hematocrit (Micro-hematocrit Capillary Tubes), (Micro Hematocrit VS-2000, Vision Scientific, Korea) 0. Hemoglobin phosphorus kit (Express plus system, Bayer, USA). 통계학적분석 (completely randomized design), SPSS version 2.0 (SPSS Inc., Chicago, IL, USA) Oneway ANOVA. Turkey s HSD (P 0.05). (mean SD) Table 2. Growth performance of olive flounder Paralichthys olivaceus (initial body weight, g) fed the experimental diets for 0 weeks Diet FBW 2 WG 3 SGR 4 FCR 5 PER 6 FI 7 SR 8 Control 32.7±3.5 ns 94±30.4 ns.56±0.5 ns.84±0.24 ns.07±0.4 ns 37.9±.8 ns 88.6±7.6 ns MCP 9 34.2±0.8 206±6.9.62±0.03.60±0.07.25±0.06 35.6±.4 90.5±4.4 DCP 9 29.4±.9 64±8.3.4±0.0.94±0.8.04±0.0 33.9±.8 90.5±.6 TCP 9 32.6±2.4 92±2.2.55±0.0.80±0.3.5±0.2 37.5±2.7 92.4±3.3 MHP 9 3.4±3.4 8±30.7.49±0.6 2.05±0.52.02±0.24 39.0±2.8 92.4±8.7 Mean values of triplicate groups are presented as mean±sd. Values in the same column having different superscript letters are significantly different (P<0.05). 2 FBW=final body weight (g/fish). 3 Weight gain (%)=00 (final mean body weight-initial mean body weight)/initial mean body weight. 4 Specific growth rate (%)=[(log e final body weight-log e initial body weight)/days] 00. 5 Feed conversion ratio=dry feed fed/wet weight gain. 6 Protein efficiency ratio=wet weight gain/total protein given. 7 Feed intake (g/fish). 8 Survival rate (%)=final fish number/initial fish number x 00. 9 MCP, monocalcium phosphate; DCP, dicalcium phosphate; TCP, tricalcium phosphate; MHP, magnesium hydrogen phosphate.

60 윤태현ㆍ이초롱ㆍ차지훈ㆍ김주민ㆍ신승준ㆍ이경준ㆍ김정대 arcsine. 결과및고찰 50% 0%, 5.3%. 6.58% (TCP) 9.90% (MCP), MHP 6.69% TCP. (P) 2.3%, 2% 2.49% (MCP) 2.54% (TCP) (Table ). 0 Table 2. 29.4 g (DCP) 32.7 g (Control). (FCR, feed conversion ratio).6 (MCP) 2.05 (MHP, magnesium hydrogen phosphate) MHP. (PER, protein efficiency ratio).02.25 (SR) 88.6 92.4% Table 3. Blood parameters of olive flounder Paralichthys olivaceus fed the experimental diets for 0 weeks Diet Hematocrit (%) Hemoglobin (g/dl) Serum inorganic Phosphorus (mg/dl) Control 24.4 ±.2 ns 4.07±0.4 ns 4.4±0.9 ns MCP 2 23.9±.6 4.04±0.4 4.7±0.7 DCP 2 23.9±.0 3.89±0.7 4.7±0.7 TCP 2 23.5±.6 3.93±0.8 4.9±0.8 MHP 2 22.9±.4 3.92±0.3 4.4±.8 Mean values of triplicate groups are presented as mean±sd. Values in the same column having different superscript letters are significantly different (P<0.05). 2 MCP, monocalcium phosphate; DCP, dicalcium phosphate; TCP, tricalcium phosphate; MHP, magnesium hydrogen phosphate.. (PCV) 22.9 24.4%, (Hb) 3.92 4.07 g/dl. 4.4 4.9 mg/dl (Table 3). Table 4. 65.3 68.7%, 4.6 6.0%.8 4.2% (P>0.05). Table 5.,,,. DCP 5.73 mg/g TCP (P<0.05)., P., (Grisdale-Helland and Helland, 998).,. Sarker et al. (2009) 270 g (Seriola quinqueradiata) K, Na Ca P 96., 94.8 92.4%. Choi et al. (2005) 4g K, Na Ca P 65, 83 85%.. 55%. P,. Hematocrit(%) Table 4. Whole body composition of olive flounder Paralichthys olivaceus (% DM) fed the experimental diets for 0 weeks Diet Protein Lipid Ash Moisture Control 68.7±3.8 ns 4.6±0.7 ns 2.9±0.6 ns 74.7±0.7 ns MCP 2 65.9±2.9 4.8±0.3 4.2±0.7 74.9±0.3 DCP 2 65.3±.5 5.3±0.7 2.6±.4 74.2±0.4 TCP 2 67.0±.0 6.0±. 2.2±.2 75.±0.4 MHP 2 65.9±0. 4.9±0.5.8±0.3 74.8±0.6 Mean values of triplicate groups are presented as mean±sd. Values in the same column having different superscript letters are significantly different (P<0.05). 2 MCP, monocalcium phosphate; DCP, dicalcium phosphate; TCP, tricalcium phosphate; MHP, magnesium hydrogen phosphate.

넙치사료내인공급제첨가효과 6 Table 5. Mineral concentration (mg/g fresh wt) of whole body composition of olive flounder Paralichthys olivaceus fed the five experimental diets for 0 weeks Diet Calcium Phosphorus Magnesium Sulfur Potassium Sodium Control 7.59±. ns 4.25±0.7 a 0.36±0.03 ns 2.28±0.0 ns 2.78±0.03 ns.00±0.03 ns MCP 2 6.70±.4 4.36±0.7 a 0.34±0.0 2.0±0.2 2.70±0.09 0.99±0.02 DCP 2 7.95±. 5.73±2. b 0.36±0.0.99±0.08 2.96±0.02 0.99±0.0 TCP 2 7.5±.0 5.4±. ab 0.32±0.04 2.32±0.40 2.70±0.05 0.97±0.05 MHP 2 7.98±.8 4.40±0.4 a 0.34±0.02.96±0.5 2.77±0.0 0.96±0.02 Mean values of triplicate groups are presented as mean±sd. Values in the same column having different superscript letters are significantly different (P<0.05). 2 MCP, monocalcium phosphate; DCP, dicalcium phosphate; TCP, tricalcium phosphate; MHP, magnesium hydrogen phosphate., Hb (g/dl) (McClatchey, 994). (Siddiqui, 977), (Barham et al., 980; Iwama et al., 986), (Giles et al., 984) (Ellsaesser and Clem, 986). PCV Hb 44-49% 8.9-0.4 g/dl (Sandnes et al., 988), 22.5-39.2% 5.2-8.3 g/dl (Vazquez and Guerrero, 2007). Rahimnejad and Lee (203) valine PCV 30.7-34.3% Hb 4.4-5.4 g/dl. Yoon et al. (206) PCV Hb 24.2-28.8% 7/5-8.8 g/ dl, Yoon et al. (207) 25.8-29.5% 7.7-8.7 g/dl. 90%., P (Yoon et al., 206; Yoon et al., 207). (Table 2), P (Table 3) (Table 4). 55%.. Kim and Shin (2006) 200 g Ca.7-.28%, P 0.5-0.57%. Kim (2000) 350 g Ca 0.62-0.76%, P 0.54-0.66%. Ca 0.84-0.86%, P 0.56-0.59% (Kim et al., 20). 30 g Ca 0.67 (MCP)-0.80% (MHP) Kim and Shin (2006) Kim (2000) Kim et al. (20). P 0.43 (Control)-0.57% (DCP)., P DCP P. (Mg), (S), (K) (Na).,,,. References AOAC (Association of Official Analytical Chemists). 995. Official Methods of Analysis. 6 th ed. Arlington, VA, U.S.A., 298. https://doi.org/0.06/0924-2244(95)90022-5. Auer MT, Kiesser MS and Canale RP. 986. Identification of critical nutrient levels through field verification of models for phosphorus and phytoplankton growth. Can J Fish Aquat Sci 43, 379-388. https://doi.org/0.39/f86-048. Barham WT, Smith GL and Schoonbee HL. 980. The hematological assessment of bacterial infection in rainbow trout, Salmo gairdneri Richardson. J Fish Biol 7, 275-28. https://doi.org/0./j.095-8649.980.tb0276.x. Choi SM, Kim KW, Kang YJ, Wang X, Kim JW, Yoo GY and Bai SC. 2005. Reevaluation of the phosphorus requirement of juvenile olive flounder Paralichthys olivaceus and the bioavailability of various inorganic phosphorus sources. J World Aquacult Soc 36, 27-222. https://doi.org/ 0./ j.749-7345.2005.tb00388.x. Ellsaesser CF and Clem LW. 986. Haematological and immunological changes in channel catfish stressed by handling and transport. J Fish Biol 28, 5-52. https://doi. org/0./j.095-8649.986.tb0587.x. Folch J, Lees M and Sloane-Stanley GH. 957. A simple method for the isolation and purification of total lipids from

62 윤태현ㆍ이초롱ㆍ차지훈ㆍ김주민ㆍ신승준ㆍ이경준ㆍ김정대 animal tissues. J Boil Chem 226, 497-509. https://dx.doi. org/0.007/bf02534078. Giles MA, Majewski HS and Hobden B. 984. Osmoregulatory and hematological responses of rainbow trout (Salmo gairdneri) to extended environmental acidification. Can J Fish Aquat Sci 4, 686-694. https://doi.org/0.39/f84-207. Grisdale-Helland B and Helland SJ. 998. Macreonutrient utilization by Atlantic halibut (Hipoglossus hipoglossus): diet digestibility and growth of kg fish. Aquaculture 66, 57-65. https://doi.org/0.06/s0044-8486(98)00274-9. Iwama GK, Greek GL and Randall DJ. 986. Changes in selected haematological parameters in juvenile chinook salmon subjected to a bacterial challenge and a toxicant. J Fish Biol 28, 563-572. https://doi.org/0./j.095-8649.986. tb0593.x. KOSIS (Korean Statistical Information Service). 207. Investigation of fisheries aquaculture production. 207. Retrieved from http://kosis.kr/stathtml/stathtml.do?mode=tab&orgi d=0&tblid=dt_ez003&vw_cd=mt_ztitle&list_ id=f38&scrid=&seqno=&lang_mode=ko&obj_var_ id=&itm_id=&conn_path=mt_ztitle&path=%252fstat isticslist%252fstatisticslistindex.do on Feb 5, 207. Kim JD, Kim KS, Song JS, Lee JY and Jeong KS. 998. Optimum level of dietary monocalcium phosphate based on growth and phosphorus excretion of mirror carp, Cyprinus carpio. Aquaculture 6, 337-344. https://doi.org/0.06/ S0044-8486(97)0028-0. Kim JD and Shin SH. 2006. Growth, feed utilization and nutrient retention of juvenile olive flounder (Paralichthys olivaceus) fed moist, semi-moist and extruded diets. Asian-Aust J Anim Sci 9, 720-726. https://doi.org/0.573/ajas.2006.720. Kim JD. 2000. Effects of dietary microbial phytase supplementation on growth and feed utilization of Japanese flounder (Paralichtys olivaceus). An Anim Res Sci, 46-59. Kim JD, Choi SM and Shin SJ. 20. Growth, Feed Utilization and Nutrient Excretions of Slow Growing Flounder (Paralichthys olivaceus) Fed either Moist or Extruded Pellet. An Anim Res Sci 22, 2-8. Kim JD. 204. Better Understanding of the Aquafeed (in Korean). The st appendix to a monthly Aquainfo magazine. Aquainfo, Seoul, Korea, 268. Kim JD. 207. Development Direction of Korean Aquafeed. In: Commemorative Seminar for the foundation of Korean Aquafeed Association. Laon Convention, Daejeon, Korea. Lall SP. 99. Digestibility, metabolism and excretion of dietary phosphorus in fish. In: Nutritional Strategies and Aquaculture Wastes. Cowey CB and Cho CY, eds. University of Guelph, Ontario, Canada, 2-36. McClatchey K. 994. Clinical laboratory medicine. Baltimore: Williams & Wilkins, Philadelphia, U.S.A. NRC (National Research Council). 20. Nutrient Requirements of Fish and Shrimp. National Academy Press, Washington, D.C., U.S.A., 376. Rahimnejad S and Lee KJ. 203. Dietary valine requirement of juvenile red sea bream Pagrus major. Aquaculture 46, 22-28. https://doi.org/0.06/j.aquaculture.203.09.026. Sandnes K, Lie Ø and Waagbø R. 988. Normal ranges of some blood chemistry parameters in adult farmed Atlantic salmon, Salmo salar. J Fish Biol 32, 29-36. https://doi. org/0./j.095-8649.988.tb0534.x. Sarker PK, Fukada H and Masumoto T. 2009. Phosphorus availability from inorganic phosphous sources in yellowtail (Seriola quinqueradiata Termminck and Schlegel). Aquaculture 289, 3-7. https://doi.org/0.06/j.aquaculture.2009.0.006. Siddiqui AQ. 977. Reproductive biology, length-weight relationship and relative condition of Tilapia leucosticte (Trewavas) in Lake Naivasha, Kenya. J Fish Biol 0, 25-260. https://doi.org/0./j.095-8649.977.tb0530.x. Vázquez GR and Guerrero GA. 2007. Characterization of blood cells and hematological parameters in Cichlasoma dimerus (Teleostei, Perciformes). Tissue and cell 39, 5-60. https:// doi.org/0.06/j.tice.2007.02.004. Yoon TH, Lee DH, Won SG, Ra CS and Kim JD. 204. Effects of dietary supplementation of magnesium hydrogen phosphate (MgHPO 4 ) as an alternative phosphorus source on growth and feed utilization of juvenile far eastern catfish (Silurus asotus). Asian-Aust J Anim Sci 27, 4-49. https://doi.org/0.573/ajas.204.4079. Yoon TH, Lee DH, Won SG, Ra CS and Kim JD. 205. Optimal incorporation level of dietary alternative phosphate (MgH- PO 4 ) and requirement for phosphorus in juvenile far eastern catfish (Silurus asotus). Asian-Aust J Anim Sci 28, -9. https://doi.org/ 0.573/ajas.4.0378. Yoon TH, Won SG, Lee DH, Choi JW, Ra CS and Kim JD. 206. Effect of a new phosphorus Source, magnesium hydrogen phosphate (MHP) on growth, utilization of phosphorus and physiological responses in Carp (Cyprinus carpio). Fish Aquat Sci 9, 49-57. https://doi.org/0.86/s4240-06-0038-4. Yoon TH, Won SG, Lee DH, Jeong GS, Choi JW, Ra CS and Kim JD. 207. Phosphorus requirement and optimum level of dietary supplementation with magnesium hydrogen phosphate (MgHPO 4 ) recovered from swine manure for Juvenile Carp Cyprinus carpio. Korean J Fish Aquat Sci 50, 46-52. http://dx.doi.org/0.5657/kfas.207.046.