JOURNAL OF FISHERIES OF CHINA ± 1. 5 g. VB mg /kg mg /kg mg /kg Lateolabrax japonicus 7 25

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1 JOURNAL OF FISHERIES OF CHINA Vol. 39 No. 4 Apr DOI /jfc * B B mg /kg ± 1. 5 g mg /kg mg /kg P < P > P < mg /kg mg /kg P < ~ 325 g mg /kg mg /kg B 1 S 963 A B 1 vitamin B mg /kg Lateolabrax japonicus 7 25 TPP mg /kg mg /kg mg /kg α α Ctenopharyngodon idella ~ mg /kg 1 Lim mg /kg B Oreochromis niloticus Oncorhynchus 2. 5 mg /kg mykiss 2-3 Ictalurus punctatus 4 Cyprinus carpio 5 Seriola quinqueradiata CARS wenhua. hb@ 163. com Oreochromis mossambicus Oreochromis niloticus 11 GIFT Oreochromis niloticus 12 5% ~ 30%

2 h r /min 10 min Tab % mg / kg mg /kg mm 4. 0 mm L ± 1. 5 g K / ~ 33 ph 7. 2 ~ mg /L 0. 5 mg /L 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 cellulose total 100 nutrients composition moisture crude protein crude fat ash mg /g mg D L- C α- E IU D IU 1 g 2. mg /g 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 α-tocopheryl acetate 50 menadione 4 retinol acetate 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 Ca CH 3 CHOHCOO 2 5H 2 O 327 FeSO MgSO 4 7H 2 O 137 NaH 2 PO NaCl AlCl 3 6H 2 O KIO KCl 75 CuCl 2 2H 2 O 0. 1 MnSO 4 H 2 O CoCl 2 6H 2 O 1 and ZnSO 4 7H 2 O 3. All ingredients were diluted with Micro-cellulose to 1 g

3 4 B 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 % = N t N 0 IBW TaKaRa SYBR Premix EX Taq TM Tli RNaseH Plus 95 5 min s s s 40 EF1 13 TK 5'-GTGGTTCGCCAGTCTGATAATG-3' 5'-CGGATGTTCTTTCCTGAG-CAG-3' EF1A 5'-GCACGCTCTGCTGGCCTTT-3' EF1A 5'-GCGCTCAATCTT-CCATCCC-3' RG QIAGEN PCR C t C t 2 - ΔΔC t SPSS 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 GBT GBT mg /kg VB mg /kg VB 1 P < mg /kg VB Sysmex Chemix P < VB mg /kg VB 1 P < TK XM VB 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 /g IBW ± 2. 2 a ± 1. 7 a ± 1. 7 a ± 2. 6 a ± 1. 4 a ± 1. 4 a /g FBW ± 7. 9 a ± 8. 9 b ± bc ± c ± 3. 3 c ± 8. 4 c /% WGR ± 3. 8 a ± 6. 9 b ± 5. 1 c ± 6. 4 c ± 6. 9 c ± 5. 6 c FE ± a ± a ± ab ± b ± b ± b /% VSI ± 0. 4 a ± 0. 3 a ± 0. 4 b ± 0. 3 b ± 0. 5 b ± 0. 2 b /% HSI ± 0. 2 a ± 0. 2 a ± 0. 2 b ± 0. 3 a ± 0. 2 a ± 0. 2 a / g /cm 3 CF ± 0. 2 a ± 0. 3 ab ± 0. 2 bc ± 0. 2 c ± 0. 1 ab ± 0. 2 ab /% SR 100 a 100 a 100 a 100 a 100 a 100 a P > Notes means in the same line sharing a same superscript letter are not significantly different determined by Duncan s test P > The same as below

4 P < % ~ 11. 4% % ~ 2. 6% mg /kg P < P > mg /kg P < mg /kg P < % mg /kg Fig. 1 Broken-line analysis of the relationship P > between dietary thiamin level and ~ 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 /% moisture ± 0. 6 a ± 0. 5 a ± 0. 6 a ± 0. 8 a ± 0. 8 a ± 0. 5 a /% crude protein ± 0. 6 a ± 0. 5 a ± 0. 3 a ± 0. 4 a ± 0. 2 a ± 0. 3 a /% crude fat ± 0. 5 a ± 0. 6 a ± 0. 5 a ± 0. 6 a ± 0. 5 a ± 0. 4 a /% ash ± a ± a ± a ± a ± a ± a 2. 3 P > mg /kg mg /kg P < P < mg /kg P < mg /kg P < Tab. 4 4 Effect of dietary thiamin level on some serum biochemical indices of GIFT tilapia / mg /kg dietary thiamin levels / mg /dl pyruvate ± b ± a ± a ± a ± a ± a / U /L ALP ± 1. 8 a ± 1. 6 ab ± 1. 3 ab ± 1. 4 c ± 1. 4 bc ± 1. 5 bc / mmol /L HDLC ± a ± b ± b ± b ± b ± b / mmol /L T-CHO ± a ± a ± b ± b ± b ± b

5 4 B P < P < P > P < mg / kg mg /kg 5 / μg /g hepatic thiamin content mg /kg 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 ± a ± b ± c ± d ± d ± d / U /kg transketolase activity ± 2. 0 a ± 3. 9 a ± 3. 9 b ± 2. 0 b ± 2. 0 b ± 2. 4 b ± a ± b ± b ± b ± b ± b transketolase expression 3 9 Scopthalmus maximus 15 Epinephelus coioides 16 Cyprinus carpio var. jian mg /kg Oncorhynchus tshawytscha g Lim g g 4 Fig. 2 Broken-line analysis of the relationship Morito 8 between dietary thiamin and hepatic thiamin concentration of GIFT tilapia mg /kg mg

6 Sheu mg /kg 19 Oncorhynchus spp. 10 ~ 15 mg /kg mg /kg mg /kg mg /kg mg /kg mg /kg mg /kg Toshiro ~ 325 g mg /kg 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 Kitamura S Suwa T Ohaara S et al. Studies on A vitamin requirements of rainbow Salmo gairdnerii The deficiency symptoms of fourth kinds of vitamin B J. Japanese Society of Fisheries Science 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 NADPH Takeshi M James W A. Thiamin requirement of α channel catfish fingerlings Ictalurus punctatus J. α Journal of Nutrition 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 Hosokawa H. The vitamin requirements of fingerling Huang 16 yellowtail Seriola quinqueradiata D. Japan Kochi University Zhong W R Zhang Sh H. Studies on the

7 4 B requirements of Lateclabrux japonicus for vitamins at various growth stages J. Journal of Zhejiang Ocean University Natural Science suppl performance functions of digestion absorption and. immune of juvenile Jian carp Cyprinus carpio var Morito C L H Conrad K H Hilton J W. The thiamin deficiency signs and requirement of rainbow trout Salmo gairdneri Richardson J. Fish Physiology and Biochemistry Halver J E. Nutrition of salmonoid fishes. III. Watersoluble 9 Zhao Z Y. Dietary myo-inositol folic acid and vitamin requirements of Chinook salmon J. thiamine requirement for grass carp Journal of Nutrition Ctenopharyngodon idellus fingerling D. Wuhan Huazhong Agricultural University Halver J E. The vitamins M Halver J E. Fish nutrition. New York Academy Press B 1 20 Masumoto T Hardy R W Casillas E. Comparison of transketolase activity and thiamin pyrophosphate 10 Lim C Yildirim-Aksoy M Barros M M et al. levels in Erythrocytes and liver of rainbow trout Thamin requirement of Nile tilapia Oreochromis Salmo gairdneri as indicators of thiamin status niloticus J. Journal of the World Aquaculture Society Lim C Leamaster B Brock J A. Thiamin requirement of red hybrid tilapia grown in seawater J. World Aquaculture Society Annual Conference and Exposition A. 12 Tendencia E A Fermin A C Dela Pe a M R et al. Effect of Epinephelus coioides Chanos chanos and GIFT tilapia in polyculture with Penaeus monodon on the growth of the luminous bacteria Vibrio harveyi J. Aquaculture Yang C G Wang X L Tian J et al. Evaluation of reference genes expression in Nile tilapia Oreochromis niloticus J. Gene Livak K J Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2 - ΔΔC t method J. Methods Cowey C B Adron J W Knox D et al. Studies on the nutrition of marine flatfish The thiamin requirement of turbot Scopthalmus maximus J. Journal of Nutrition Huang J W Tian L X Du Z Y et al. Effects of dietary thiamin on the physiological status of the grouper Epinephelus coioides J. Fish Physiology and Biochemistry Huang H H. Effect of thiamin on growth Jian D. Yaan Sichuan Agricultural University J. Journal of Nutrition McCourt J A Nixon P F Duggleby R G. Thiamin nutrition and catalysis-induced instability of thiamin diphosphate J. British Journal of Nutrition Ma K Hutchins A Sung S J et al. Pyruvate ferredoxin oxidoreductase from the hyperthermophilic archaeon Pyrococcus furiosus functions as a CoA-dependent pyruvate decarboxylase J. Proceedings of the National Academy of Sciences of the United States of America Zhang J Xiong Z Y. The effect of the exercise performance with the lack of vitamin B 1 J. Journal of Ankang Teachers College B Rindi G. Thiamin C Ziegler E E Filer L J. Present Knowledge in Nutrition. Washington DC International Life Sciences Institute Press Sheu K F Calingasan N Y Dienel G A et al. Regional reductions of transketolase in thiaminedeficient rat brain J. Journal of Neurochemistry

8 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 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 China Abstract GIFT tilapia initial weight ± 1. 5 g were fed purified diets containing six levels and 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 < 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 < while protein moisture ash lipid ontent in whole body had no significant differences among various treatments P > 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 and mg /kg diet respectively. Key words GIFT Oreochromis niloticus thiamin B 1 requirement growth Corresponding author WEN Hua. wenhua. hb@ 163. com

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