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39 4 2015 4 JOURNAL OF FISHERIES OF CHINA Vol. 39 No. 4 Apr. 2015 1000-0615 2015 04-0539 - 08 DOI 10. 11964 /jfc. 20140909476 1 2* 2 1 2 2 2 2 2 2 B 1 1. 430070 2. 430223 B 1 0. 08 0. 57 1. 13 2. 09 4. 11 8. 09 mg /kg 6 64. 4 ± 1. 5 g 12 1. 13 2. 09 4. 11 8. 09 mg /kg 2. 09 mg /kg P < 0. 05 P > 0. 05 P < 0. 05 1. 13 mg /kg 0. 57 mg /kg P < 0. 05 64 ~ 325 g 1. 16 mg /kg 2. 06 mg /kg B 1 S 963 A B 1 vitamin B 1 1. 0 mg /kg Lateolabrax japonicus 7 25 TPP mg /kg 8 1. 0 mg /kg 5 0. 5 mg /kg α α Ctenopharyngodon idella 9 1. 16 ~ 4. 49 mg /kg 1 Lim 10 3. 5 mg /kg B Oreochromis niloticus Oncorhynchus 2. 5 mg /kg mykiss 2-3 Ictalurus punctatus 4 Cyprinus carpio 5 Seriola quinqueradiata 6 4 2014-09-18 2014-11-22 CARS-49 201003020 E-mail wenhua. hb@ 163. com Oreochromis mossambicus Oreochromis niloticus 11 GIFT Oreochromis niloticus 12 5% ~ 30%

540 39 4 2 h 3 000 r /min 10 min - 40 3-80 1 Tab. 1 1 1. 1 98% 0 0. 5 1. 0 2. 0 4. 0 8. 0 mg / kg 6 6 0. 08 0. 57 1 1. 13 2. 09 4. 11 8. 09 mg /kg 60 2. 0 mm 4. 0 mm - 20 1. 2 500 L 2 64. 4 ± 1. 5 g 18 15 6 3 6 9 00 16 00 2 1 50 K 12 11 00 1 /3 125 27 ~ 33 ph 7. 2 ~ 7. 5 5. 0 mg /L 0. 5 mg /L 1. 3 12 24 h 6 3 H 2 O 3 6H 2 O Formulation and proximate analysis of the basal diet ingredient /% content casein 30 gelatin 7. 5 dextrin 38 corn oil 4 soy oil 4 mineral premix 4 vitamin premix 1 choline chloride 0. 25 cellulose 11. 25 total 100 nutrients composition moisture 9. 41 crude protein 32. 23 crude fat 6. 78 ash 2. 66 1. mg /g 10 10 mg D- 0. 6 4 1. 5 200 L- C-2-60 6. 05 α- E 4 2 000 IU D 3 400 IU 1 g 2. mg /g 135. 8 327 2. 137 87. 2 43. 5 0. 15 0. 125 75 0. 1 0. 80 1 3 1 g Notes 1. the vitamin mixture supplied the following mg /g mixture riboflavin 10 calcium Pantothenate 10 mg D-biotin 0. 6 Pyridoxine hydrochloride 4 folic acid 1. 5 inositol 200 L-ascorbyl- 2-monophosphate-Mg 60 nicotinic acid 6. 05 α-tocopheryl acetate 50 menadione 4 retinol acetate 2 000 IU and cholecaliferol 400 IU. All ingredients were diluted with Micro-cellulose to 1 g. 2. the mineral mixture supplied the following mg /g diet Ca H 2 PO 4 2 135. 8 Ca CH 3 CHOHCOO 2 5H 2 O 327 FeSO 4 2. 125 MgSO 4 7H 2 O 137 NaH 2 PO 4 87. 2 NaCl 43. 5 AlCl 3 6H 2 O 0. 15 KIO 3 0. 125 KCl 75 CuCl 2 2H 2 O 0. 1 MnSO 4 H 2 O 0. 80 CoCl 2 6H 2 O 1 and ZnSO 4 7H 2 O 3. All ingredients were diluted with Micro-cellulose to 1 g

4 B 1 541 survival rate SR % = 100 N t /N 0 1 weight gain rate WGR % = 100 FBW - IBW /IBW 2 feed efficiency FE = FTBW - ITBW /FG L 3 W VW /W 3 condition factor CF g /cm 3 =100 W / 4 hepatosomatic HSI % = 100 HW / 5 viscerosomatic index VSI % = 100 6 N t N 0 IBW TaKaRa SYBR Premix EX Taq TM Tli RNaseH Plus 95 5 min 95 15 s 60 15 s 72 20 s 40 EF1 13 TK 5'-GTGGTTCGCCAGTCTGATAATG-3' 5'-CGGATGTTCTTTCCTGAG-CAG-3' EF1A 5'-GCACGCTCTGCTGGCCTTT-3' EF1A 5'-GCGCTCAATCTT-CCATCCC-3' RG- 6000 QIAGEN PCR C t C t 2 - ΔΔC t 14 1. 4 SPSS 19. 0 One-Way ANOVA g FBW g W Duncan g L cm HW g VW ± P <0. 05 g FG ITBW 2 g FTBW g 2. 1 VB 105 1 12 GBT 147002002 GBT 9695. 27-2008 1. 13 2. 09 4. 11 8. 09 mg /kg VB 1 0. 08 0. 57 mg /kg VB 1 P < 0. 05 0. 57 mg /kg VB Sysmex Chemix-800 1 P < 0. 052 VB 1 2. 09 4. 11 8. 09 mg /kg VB 1 P < 0. 05 TK XM VB -005477993 1 1. 13 2. 09 4. 11 8. 09 mg /kg VB 1 2 Tab. 2 Effect of dietary thiamin supplement on the growth performance feed efficiency and body indices of GIFT tilapia / mg /kg dietary thiamin levels 0. 08 0. 57 1. 13 2. 09 4. 11 8. 09 /g IBW 64. 3 ± 2. 2 a 64. 2 ± 1. 7 a 63. 9 ± 1. 7 a 64. 5 ± 2. 6 a 64. 8 ± 1. 4 a 64. 7 ± 1. 4 a /g FBW 282. 4 ± 7. 9 a 302. 2 ± 8. 9 b 318. 1 ± 11. 4 bc 325. 0 ± 12. 3 c 321. 8 ± 3. 3 c 325. 8 ± 8. 4 c /% WGR 339. 6 ± 3. 8 a 370. 9 ± 6. 9 b 397. 8 ± 5. 1 c 403. 5 ± 6. 4 c 396. 8 ± 6. 9 c 402. 3 ± 5. 6 c FE 0. 74 ± 0. 07 a 0. 72 ± 0. 05 a 0. 84 ± 0. 05 ab 0. 91 ± 0. 06 b 0. 90 ± 0. 03 b 0. 92 ± 0. 07 b /% VSI 8. 38 ± 0. 4 a 8. 60 ± 0. 3 a 9. 29 ± 0. 4 b 9. 21 ± 0. 3 b 9. 26 ± 0. 5 b 9. 17 ± 0. 2 b /% HSI 2. 36 ± 0. 2 a 2. 36 ± 0. 2 a 3. 03 ± 0. 2 b 2. 57 ± 0. 3 a 2. 59 ± 0. 2 a 2. 52 ± 0. 2 a / g /cm 3 CF 3. 52 ± 0. 2 a 3. 68 ± 0. 3 ab 3. 90 ± 0. 2 bc 4. 14 ± 0. 2 c 3. 68 ± 0. 1 ab 3. 60 ± 0. 2 ab /% SR 100 a 100 a 100 a 100 a 100 a 100 a P > 0. 05 Notes means in the same line sharing a same superscript letter are not significantly different determined by Duncan s test P > 0. 05. The same as below

542 39 P < 0. 05 10. 6% ~ 11. 4% 1. 13 2. 5% ~ 2. 6% mg /kg P < 0. 05 P > 0. 05 2. 09 mg /kg P < 0. 05 1. 13 mg /kg P < 0. 05 100% 1. 16 mg /kg 1 1 2. 2 Fig. 1 Broken-line analysis of the relationship P > 0. 05 between dietary thiamin level and 3 69. 0 ~ weight gain rate of GIFT tilapia 70. 5% 15. 0% ~ 15. 9% Tab. 3 3 Effect of dietary thiamin levels on body composition of GIFI tilapia / mg /kg dietary thiamin levels 0. 08 0. 57 1. 13 2. 09 4. 11 8. 09 /% moisture 69. 6 ± 0. 6 a 69. 9 ± 0. 5 a 69. 0 ± 0. 6 a 69. 1 ± 0. 8 a 69. 9 ± 0. 8 a 70. 5 ± 0. 5 a /% crude protein 15. 1 ± 0. 6 a 15. 0 ± 0. 5 a 15. 6 ± 0. 3 a 15. 9 ± 0. 4 a 15. 1 ± 0. 2 a 15. 3 ± 0. 3 a /% crude fat 11. 4 ± 0. 5 a 10. 6 ± 0. 6 a 11. 6 ± 0. 5 a 11. 5 ± 0. 6 a 10. 9 ± 0. 5 a 11. 0 ± 0. 4 a /% ash 2. 62 ± 0. 09 a 2. 50 ± 0. 08 a 2. 49 ± 0. 09 a 2. 62 ± 0. 08 a 2. 49 ± 0. 07 a 2. 64 ± 0. 09 a 2. 3 P > 0. 05 2. 09 4. 11 8. 09 mg /kg 0. 08 0. 57 1. 13 mg /kg P < 0. 05 4 P < 0. 05 1. 13 2. 09 4. 11 8. 09 mg /kg P < 0. 05 0. 08 0. 57 mg /kg P < 0. 05 Tab. 4 4 Effect of dietary thiamin level on some serum biochemical indices of GIFT tilapia / mg /kg dietary thiamin levels 0. 08 0. 57 1. 13 2. 09 4. 11 8. 09 / mg /dl pyruvate 1. 09 ± 0. 02 b 0. 55 ± 0. 03 a 0. 52 ± 0. 02 a 0. 50 ± 0. 02 a 0. 52 ± 0. 02 a 0. 50 ± 0. 01 a / U /L ALP 18. 8 ± 1. 8 a 21. 3 ± 1. 6 ab 20. 8 ± 1. 3 ab 24. 7 ± 1. 4 c 22. 0 ± 1. 4 bc 22. 4 ± 1. 5 bc / mmol /L HDLC 0. 62 ± 0. 06 a 1. 04 ± 0. 06 b 0. 93 ± 0. 08 b 0. 93 ± 0. 07 b 0. 91 ± 0. 06 b 1. 02 ± 0. 09 b / mmol /L T-CHO 3. 60 ± 0. 60 a 3. 68 ± 0. 40 a 5. 01 ± 0. 52 b 4. 84 ± 0. 43 b 4. 66 ± 0. 23 b 4. 93 ± 0. 34 b

4 B 1 543 2. 4 P < 0. 05 0. 57 1. 13 2. 09 4. 11 8. 09 P < 0. 05 P > 0. 05 P < 0. 055 1. 13 2. 09 4. 11 8. 09 mg / kg 0. 08 0. 57 mg /kg 5 / μg /g hepatic thiamin content mg /kg 2. 06 mg /kg 2 Tab. 5 Effect of dietary thiamin levels on hepatic thiamin transketolase activity transketolase expression of GIFT tilapia / mg /kg dietary thiamin levels 0. 08 0. 57 1. 13 2. 09 4. 11 8. 09 0. 34 ± 0. 02 a 0. 45 ± 0. 01 b 0. 59 ± 0. 02 c 0. 75 ± 0. 03 d 0. 76 ± 0. 03 d 0. 76 ± 0. 03 d / U /kg transketolase activity 45. 0 ± 2. 0 a 45. 7 ± 3. 9 a 61. 4 ± 3. 9 b 63. 9 ± 2. 0 b 68. 2 ± 2. 0 b 65. 6 ± 2. 4 b 1. 00 ± 0. 23 a 1. 60 ± 0. 13 b 1. 78 ± 0. 13 b 1. 79 ± 0. 12 b 1. 81 ± 0. 13 b 1. 89 ± 0. 21 b transketolase expression 3 9 Scopthalmus maximus 15 Epinephelus coioides 16 Cyprinus carpio var. jian 17 11 2. 5 mg /kg Oncorhynchus tshawytscha 18 5 10 6 8 64 g Lim 10 2 2 5. 30 g 6 3. 62 g 4 Fig. 2 Broken-line analysis of the relationship Morito 8 between dietary thiamin and hepatic thiamin concentration of GIFT tilapia 17 4 8 11 1. 16 mg /kg 10 3. 5 mg

544 39 Sheu 25 7 25 mg /kg 19 Oncorhynchus spp. 10 ~ 15 mg /kg 6 11. 2 mg /kg 9 1. 16 mg /kg 17 1. 02 mg /kg 8 1. 0 mg /kg 4 1. 0 mg /kg 5 0. 5 mg /kg 4 1 9 Toshiro 20 64 ~ 325 g 1. 16 2. 06 mg /kg 2. 06 mg /kg 1 NRC National Research Council. Vitamins M Nutrient requirements of fish and shrimp. Washington DC USA National Academy Lim 10 Morito 8 Press 2011. 2 Kitamura S Suwa T Ohaara S et al. Studies on A vitamin requirements of rainbow Salmo gairdnerii. 21-23 2. The deficiency symptoms of fourth kinds of vitamin B J. Japanese Society of Fisheries Science 1967 33 12 1120-1125. 3 Lehmitz R Spannhof L. Transketolase activity and thiamin deficiency of the kidney in rainbow trout Salmo gairdneri following contionous feeding with raw fearing J. Arch Tierernahr 1977 27 4 287 - NADPH 24 295. 4 Takeshi M James W A. Thiamin requirement of α channel catfish fingerlings Ictalurus punctatus J. α Journal of Nutrition 1978 108 1 176-180. 5 Aoe H Masuda I Mimura T et al. Water-soluble 15 vitamin requirements of carp-vi. Requireement for Masumoto 20 mrna thiamine and effects of antithiamines J. The Japanese Society of Fisheries Science 1969 35 5 459-465. 6 Hosokawa H. The vitamin requirements of fingerling Huang 16 yellowtail Seriola quinqueradiata D. Japan Kochi University 1989. 7 Zhong W R Zhang Sh H. Studies on the

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546 39 Dietary thiamin requirement of GIFT Oreochromis niloticus REN Chun 1 WEN Hua 2* HUANG Feng 2 JIANG Ming 1 2 WU Fan 2 LIU Wei 2 TIAN Juan 2 YANG Changgeng 2 WU Jinping 2 1. College of Fisheries Huazhong Agricultural University Wuhan 430070 China 2. Key laboratory of Freshwater Biodiversity Conservation and Utilization of Ministry of Agriculture Yangtze River Fisheries Research Institute Chinese Academy of Fishery Sciences Wuhan 430223 China Abstract GIFT tilapia initial weight 64. 4 ± 1. 5 g were fed purified diets containing six levels 0. 08 0. 57 1. 13 2. 09 4. 11 and 8. 09 mg /kg diet respectively of thiamin for 12 weeks. The experiment was used to determine the dietary thiamin requirement of GIFT tilapia on the basis of growth parameters hepatic thiamin saturation and a thiamin-dependent biochemical function. The results showed that w ith the increasing dietary thiamin level w eight gain rate of tilapia first increased linearly then remained nearly unchanged. Hepatic thiamin contents presented the same tendency w ith w eight gain rate. Fish fed diets supplemented w ith showed significantly higher hepatic transketolase expression serum high density lipoprotein cholesterol and lower serum pyruvate than fish fed diets without supplementation of P < 0. 05. Fish fed the control diet w ithout supplementation of thiamin show ed significantly higher serum pyruvate content than fish fed diets supplemented with thiamin P < 0. 05 while protein moisture ash lipid ontent in whole body had no significant differences among various treatments P > 0. 05. Based on broken method of the relationships between weight gain rate hepatic thiamin contents and dietary concentrations it can be concluded that the dietary requirement for GIFT tilapia was considered to be 1. 16 and 2. 06 mg /kg diet respectively. Key words GIFT Oreochromis niloticus thiamin B 1 requirement growth Corresponding author WEN Hua. E-mail wenhua. hb@ 163. com