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1 2016 年 9 月, 23(5): Journal of Fishery Sciences of China 研究论文 DOI: /SP.J 吉富罗非鱼对饲料中苯丙氨酸的需要量蒋明 1 1,, 武文一 2, 文华 1, 刘伟 1, 吴凡 1, 田娟 1 1, 杨长庚 1.,, ; 2., 摘要 : 6 ( 0.78% 0.95% 1.09% 1.34% 1.51% 1.72%) (phenylalanine, Phe) ( 30.10%, MJ/kg), ( 24~32 ) ( ) g 60 d, Phe (GIFT, Oreochromis niloticus),, Phe,, ; (P<0.05), (P<0.05) Phe (P>0.05); (P>0.05) Phe, (P<0.05), (P>0.05); Phe Na + -K + -ATP (P<0.05), (P>0.05),, Phe 1.17%~1.21%, 3.89%~4.02% 关键词 : ; ; ; 中图分类号 : S963 文献标志码 : A 文章编号 : (2016) ,,,, [1] (GIFT, Oreochromis niloticus), [2],, 10 [3], (phenylalanine, Phe), [3] Phe (Tyrosine, Tyr) Tyr,,, [4] Phe Tyr, Tyr Phe, Phe Tyr, Phe [5], Tyr,, Phe [6], Phe,,, 收稿日期 : ; 修订日期 : 基金项目 : (CARS-49); ( ) ( ). 作者简介 : (1979 ),,,. jiangm.hb@163.com 通信作者 :,. wenhua.hb@163.com

2 [7] ; Phe (Ctenopharyngodon idella) [6] 鲃 (Catla catla) [8] (Chanos chanos Forsskal) [9] (Carassius auratus gibelio) [10] (Megalobrama amblycephala) [11], (20 g) 1.57% [12], Santiago [13] (0.013~1 g) Phe 1.05%( Tyr 0.5%),, [14],, Phe 60 d,, Phe, 1 材料与方法 1.1 试验饲料 ( α- 1 : 1 : 1 ),, ( Phe ) [15], Millamena [16], ( 1), Phe 0%( ), 0.20%, 0.40%, 0.60%, 0.80% 1.00%(L- ) 80, 1,, 2 mm, 60 5 mm, 20 Phe 0.78%, 0.95%, 1.09%, 1.34%, 1.51% 1.72%, 2.59%, 3.16%, 3.62%, 4.45%, 5.02% 5.71% 2 composition 表 1 实验饲料组成及主要营养水平 ( 风干基础 ) Tab. 1 Composition and nutrient levels of test diets (air-dry basis) % item fish meal gelatin peanut soybean meal amino acid mixture dextrin soybean oil corn oil choline chloride ) vitamin premix 1) ) mineral premix 2) antiseptic BHT bentonite microcrystalline L- L-Gly L- L-Phe total ( 待续 to be continued)

3 5 : 1175 proximate composition ( 续表 1 Tab. 1 continued) item /(MJ kg 1 ) gross energy crude protein crude lipid ash : 1) kg : A, 5000 IU; D 3, 2000 IU; E, 60 mg; C, 120 mg; K 3, 5 mg; B 1, 5 mg; B 2, 20 mg; B 6, 10 mg;, 120 mg;, 10 mg;, 1 mg;, 0.1 mg;, 400 mg. 2) kg : Ca(H 2 PO 4 ) 2, mg; Ca(CH 3 CHOHCOO) 2, 6540 mg; FeSO 4, 42.5 mg; MgSO 4, 1340 mg; NaH 2 PO 4, 1744 mg; NaCl, 870 mg; AlCl 3, 3 mg; KIO 3, 2.5 mg; KCl, 1500 mg; CuCl 2, 2 mg; MnSO 4, 16 mg; CoCl 2, 20 mg; ZnSO 4, 60 mg. Note: 1) The premix provided the following per kg of diets: vitamin A, 5000 IU; vitamin D 3, 2000 IU; vitamin E, 60 mg; vitamin C, 120 mg; vitamin K 3, 5 mg; vitamin B 1, 5 mg; vitamin B 2, 20 mg; vitamin B 6, 10 mg; nicotinic acid, 120 mg; calcium pantothenate, 10 mg; folic acid, 1 mg; biotin, 0.1 mg; inositol 400 mg. 2) The premix provided the following per kg of diets: Ca(H 2 PO 4 ) 2, mg; Ca(CH 3 CHOHCOO) 2, 6540 mg; FeSO 4, 42.5 mg; MgSO 4, 1340 mg; NaH 2 PO 4, 1744 mg; NaCl, 870 mg; AlCl 3, 3 mg; KIO 3, 2.5 mg; KCl, 1500 mg; CuCl 2, 2 mg; MnSO 4, 16 mg; CoCl 2, 20 mg; ZnSO 4, 60 mg. 表 2 基础饲料中氨基酸组成分析 Tab. 2 Analyzed amino acid composition of the basal diet % % dry matter 30% ( ) provided by provided by amount in 30% muscle amino acid AA in basal diet(measured value) fish meal-gelatin-peanut meal-soybean meal AA mixture protein Met Lys Thr Try ND * Arg Ile Leu Val His Phe Cys Gly Tyr Ser Pro Ala Asp Glu : *. Note: * Try was not detected. 1.2 试验鱼与饲养管理, (3%NaCl ), (4.0 m 4.0 m 1.5 m); 15 d,, 24 h, 270 [ (52.70±1.80) g], 18 (1.0 m 1.0 m 1.5 m), 15, 6, 3, 6, 60 d, 1 24~32, ph 6.5~7.0, 4 mg/l,

4 mg/l 2 ( 8:00; 16:00),, 7000 m 2, 3 m, 3 kw,, 1: 00 4: 00, 2 h,, 4 mg/l, 1.3 样品采集, 24 h;, ; 3, 3, 30 mg/l (MS-222),,, 4 2 h, 3000 r/min 15 min,,,, ; 40 ; (4 ) (4, 4000 r/min)15 min, 40, 1.4 指标测定, (weight gain rate) (specific growth rate) (feed conversion ratio) (protein efficiency rate) (protein deposition rate) (survival rate) (condition factor) (hepatosomatic index) (viscerosomatic index) (WGR, %) (W t W 0 ) 100/W 0 ; (SGR, % d 1 )= (lnw t lnw 0 ) 100/t (FCR)=F/(W t W 0 ) (PER, %)=(W t W 0 )/F P (PDR, %) =100 (W t P t W 0 P 0 )/F P (CF, g/cm 3 )=W 100/L 3 (HSI, %)=W h 100/W (VSI, %)=W v 100/W, N t, N 0 ; W t (g), W 0 (g), W (g); t (d); F (g); P (%), P t (%), P 0 (%), W h (g), W v (g), L (cm) 105, ;,, (Parr-6200), L-8900 ( GB/T ) (Sysmex, CHEMIX- 800) (AST) (ALT) (TG) (T-CHO) (GLU) Na + -K + -ATP 1.5 数据处理 ± ( x ±SD), SPSS 18.0 One-Way ANOVA Tukey s, P<0.05, 2 结果与分析 2.1 饲料苯丙氨酸水平对吉富罗非鱼生长性能和饲料蛋白质利用率的影响 3, Phe (FBW) (WGR) (SGR) (FCR) (PER) (PDR) (CF) (HSI) (VSI)(P<0.05) Phe, WGR SGR PER PDR 1.34%, WGR SGR PER PDR, (P<0.05) Phe, FCR, 1.34%

5 5 : 1177, (P<0.05) Phe (1.72% ) CF HSI VSI Phe ; 1.34%, 0.95 % 1.09% 1.51% (P>0.05), (P<0.05); 1.09%, 0.95% 1.34% 1.51% (P>0.05), (P<0.05) Tab. 3 表 3 饲料苯丙氨酸水平对罗非鱼生长性能饲料利用和形态学指标的影响 Effects of dietary phenylalanine levels on growth performance, feed utilization and morphometry indexes of GIFT, Oreochromis niloticus n=3; x ±SD item /g IBW 53.56± ± ± ± ± ±0.67 /g FBW ±1.94 c ±1.41 b ±1.60 a ±1.09 a ±1.88 c ±1.44 d /% WGR ±1.17 c ±1.74 b ±2.10 a ±2.20 a ±1.27 c ±2.47 d /(% d 1 ) SGR 2.53±0.01 c 2.57±0.01 b 2.60±0.01 a 2.61±0.01 a 2.53±0.02 c 2.45±0.01 d FCR 1.37±0.01 c 1.33±0.01 b 1.25±0.01 a 1.26±0.01 a 1.35±0.02 c 1.42±0.01 d /% PER 2.43±0.02 c 2.50±0.02 b 2.65±0.01 a 2.66±0.01 a 2.46±0.03 c 2.34±0.02 d /% PDR 38.44±0.10 c 39.99±0.10 b 40.88±0.17 a 40.91±0.17 a 39.27±0.32 b 38.50±0.28 d /[g cm 3 ]CF 3.53±0.23 b 3.83±0.27 a 3.97±0.25 a 3.68±0.32 ab 3.75±0.18 ab 3.48±0.32 b /% HSI 1.88±0.58 bc 1.99±0.59 abc 2.38±0.53 ab 2.49±0.64 a 2.02±0.43 abc 1.75±0.36 c /% VSI 9.70±0.84 ab 9.69±1.30 ab 10.75±1.10 a 10.07±1.54 a 9.66±0.93 ab 8.68±1.11 b : (P>0.05, Tukey s ); (P<0.05). Notes: Values in the same line sharing the same superscript letter are not significantly different determined by Tukey s test (P>0.05), and values with different letters are significantly different (P<0.05). Phe (X), WGR(Y 1 ) FCR(Y 2 ) PER(Y 3 ), ( 1 3) : Y 1 = X X (R 2 =0.9523), X=1.17; Y 2 =0.6529X X (R 2 =0.95), X=1.21; Y 3 = X X (R 2 =0.9367), X= Fig. 2 The relationship between feed conversion ratio and dietary phenylalanine levels of GIFT, Oreochromis niloticus 1 Fig. 1 The relationship between weight gain rate and dietary phenylalanine levels of GIFT, Oreochromis niloticus 2.2 饲料苯丙氨酸水平对吉富罗非鱼体成分和肌肉氨基酸含量的影响 4, Phe

6 (P<0.05), (P>0.05) Phe,, 1.34% ;, 1.09% 5, Phe (P>0.05) 2.3 饲料苯丙氨酸水平对吉富罗非鱼血清生化指标的影响 6, Phe (ALT), (TG) (T-CHO) (GLU) (P<0.05) 3 Fig. 3 The relationship between protein efficiency rate and dietary phenylalanine levels of GIFT, Oreochromis niloticus whole fish Tab. 4 表 4 饲料苯丙氨酸水平对罗非鱼全鱼和肌肉组成的影响 Effect of dietary phenylalanine levels on body and muscle composition of GIFT, Oreochromis niloticus n=3; x ±SD; % item moisture 69.26± ± ± ± ± ±0.32 crude protein 15.32± ± ± ± ± ±0.39 crude lipid 9.41±0.85 b 9.46±0.40 b 9.69±0.48 b 11.24±0.97 a 11.17±0.23 a 10.23±0.78 ab ash 3.10±0.22 a 4.25±0.35 c 3.50±0.17 ab 4.02±0.56 bc 3.58±0.37 ab 3.78±0.33 bc muscle moisture 76.54± ± ± ± ± ±0.35 crude protein 19.43± ± ± ± ± ±1.33 crude lipid 1.54± ± ± ± ± ±0.45 ash 1.29±0.04 ab 1.32±0.08 ab 1.21±0.05 b 1.29±0.08 ab 1.37±0.09 ab 1.41±0.17 a : (P>0.05, Tukey s ); (P<0.05). Notes: Values in the same line sharing the same superscript letter are not significantly different determined by Tukey s test (P>0.05), and values with different letters are significantly different (P<0.05). 表 5 饲料苯丙氨酸水平对吉富罗非鱼肌肉氨基酸含量 ( 干物质 ) 的影响 Tab. 5 Effect of dietary phenylalanine levels on amino acid composition (dry matter) of GIFT, Oreochromis niloticus n=3; x ±SD; % amino acid Met 2.50± ± ± ± ± ±0.14 Lys 7.70± ± ± ± ± ±0.50 Thr 3.84± ± ± ± ± ±0.15 Arg 5.05± ± ± ± ± ±0.26 Ile 3.89± ± ± ± ± ±0.24 Leu 6.67± ± ± ± ± ±0.38 Val 3.99± ± ± ± ± ±0.25 His 2.28± ± ± ± ± ±0.17 Phe 3.61± ± ± ± ± ±0.17 ( 待续 to be continued)

7 5 : 1179 ( 续表 5 Tab. 5 continued) amino acid Tyr 2.70± ± ± ± ± ±0.01 Cys 0.82± ± ± ± ± ±0.04 Gly 4.81± ± ± ± ± ±0.28 Ser 3.34± ± ± ± ± ±0.14 Pro 2.81± ± ± ± ± ±0.17 Ala 5.06± ± ± ± ± ±0.23 Asp 8.77± ± ± ± ± ±0.45 Glu 12.38± ± ± ± ± ±0.60 EAA 80.22± ± ± ± ± ±4.01 TAA 39.53± ± ± ± ± ±2.24 :. Note: Try was not detected. Tab. 6 表 6 饲料苯丙氨酸水平对吉富罗非鱼血清生化指标的影响 Effect of dietary phenylalanine levels on serum biochemical parameters of GIFT, Oreochromis niloticus n=3; x ±SD; % item /(U L 1 ) AST 98.25± ± ± ± ± ±1.01 /(U L 1 ) ALT 21.50±9.06 ab 17.25±4.50 ab 12.00±3.32 b 15.50±2.94 ab 23.50±9.38 ab 27.75±7.97 a /(mmol L 1 ) T-CHO 3.89±0.37 b 3.98±1.11 b 5.27±0.66 a 4.26±0.49 ab 4.38±0.70 ab 4.52±0.97 ab /(mmol L 1 ) GLU 3.13±1.01 b 2.42±1.06 b 5.13±0.72 a 4.98±1.88 a 3.43±0.85 ab 3.01±0.75 b /(mmol L 1 ) TG 2.71±0.31 b 3.79±2.38 ab 6.59±1.20 a 3.41±1.87 ab 3.70±1.39 ab 2.47±1.21 b : (P>0.05, Tukey s ); (P<0.05). Notes: Values in the same line sharing the same superscript letter are not significantly different determined by Tukey s test (P>0.05), and values with different letters are significantly different (P<0.05). 1.09%, (P<0.05);, 1.09%, (P<0.05); (P>0.05) 2.4 饲料苯丙氨酸水平对罗非鱼前肠消化酶活性的影响 Phe ( 7) Phe, Na + -K + - ATP Tab. 7 表 7 饲料苯丙氨酸水平对吉富罗非鱼前肠消化酶活性的影响 Effect of dietary phenylalanine levels on digestive enzyme activities in the intestine of GIFT, Oreochromis niloticus n=3; x ±SD; % item Na + -K + -ATP /(U mg 1 ) intestinal Na + -K + -ATPase 1.01±0.32 b 1.52±0.26 ab 1.21±0.43 ab 1.41±0.49 ab 1.44±0.31 ab 1.71±0.19 a /(U mg 1 ) intestinal protease 6.16±0.62 b 11.68±5.18 ab 19.07±9.83 ab 16.25±5.42 ab 24.55±18.47 a 22.90±6.94 a /(U g 1 ) intestinal lipase 83.56±5.55 c ±13.82 b ±16.38 ab ±13.07 a ±17.49 a ±18.74 ab /(U mg 1 ) intestinal amylase 0.18± ± ± ± ± ±0.04 : (P>0.05, Tukey s ); (P<0.05). Notes: Values in the same line sharing the same superscript letter are not significantly different determined by Tukey s test (P>0.05), and values with different letters are significantly different (P<0.05).

8 , (P>0.05) Phe 1.72%, Na + -K + -ATP (P<0.05); 1.51%, (P<0.05), % % 3 讨论 3.1 饲料苯丙氨酸水平对吉富罗非鱼生长性能和饲料蛋白质利用率的影响 Phe, [3] Phe (Oncorhynchus mykiss) [17] (Bidyanus bidyanus) [18], Phe, [6] [11], Phe,, Phe, Phe,,, [10] Phe,, [19],, Phe,,,, Phe 1.17%~1.21%( 3.89%~4.02%) (0.013~1 g) Phe (1.05%) [13], 20% [20], Phe ; [10] (1.09%) [11] (0.93%~ 1.00%) [6] (1.04%) [21] (Salmo salar) (0.77%) 鮰 (Ictalurus punctatus) [22] (0.6%), 鲃 (Barbus capito) [23] (1.70%) (Bidyanus bidyanus) [18] (2.20%) [24] (1.27%); (Acipenser transmontanus) (1.19%) [25] (Oncorhynchus mykiss) (1.16%) [17] Phe [26], Phe [7] Phe 3.2 饲料苯丙氨酸水平对吉富罗非鱼体组成的影响 Phe Khan [7] 鲃 (Catla catla), Phe Phe, Phe, Phe, [10] (Labeo rohita) [7],, Phe, [5], Phe,, Phe ( ) [27], 3.3 饲料苯丙氨酸水平对吉富罗非鱼血清生化指标的影响 [28], (ALT) (TG) (T-CHO) (GLU) Phe (ALT)

9 5 : 1181, [29] Phe ALT, Phe, Phe,,, ALT [30] ;, Phe AST Phe, GLU 1.09%, Phe [11] GLU ;,,, T-CHO TG, GLU, Phe GLU [31], Zhou [32] (Rachycentron canadum), ; 3.4 饲料苯丙氨酸水平对吉富罗非鱼前肠消化酶活性的影响 [33],, [34] Phe,,,,, [35] Phe,,, [36] Na + -K + - ATP, Na + -K +, [37], Na + -K + -ATP, Na +, Na +, Na + -K + -ATP [38] Na + -K + -ATP Phe, Phe Na + -K + -ATP, 4 结论 Phe,,,, Phe 1.17%~1.21%, 3.89%~ 4.02% 参考文献 : [1] Kanshik S J, Seiliez I. Protein and amino acid nutrition and metabolism in fish: Current knowledge and future needs[j]. Aquac Res, 2010, 41(3): [2] El-Sayed A F M. Tilapia feed and feeding in semi-intensive culture systems[c]//from the pharaohs to the future. Eighth International Symposium on Tilapia in Aquaculture Proceedings. Cairo, Egypt, 2008: [3] National Research Council(NRC). Nutrient Requirements of Fish and Shrimp[M]. Washington, D C: National Academies Press, 2011, 69 70: [4] Zhou S W. Animal Biochemistry[M]. Beijing: China Agriculture Press, 1999: [. [M]. :, 1999: ] [5] He Z G, Mai K S, Ai Q H. The recent advance on phenylalanine nutritional physiology in fish[j]. China Feed, 2012(5): [,,. [J]., 2012(5): ] [6] Li W, Feng L, Liu Y, et al. Effects of dietary phenylalanine on growth, digestive and brush border enzyme activities and antioxidant capacity in the hepatopancreas and intestine of young grass carp (Ctenopharyngodon idella)[j]. Aquac Nutr, 2015, 21(6): [7] Khan M A, Abidi S F. Total aromatic amino acid requirement of Indian major carp Labeo rohita (Hamilton) fry[j]. Aquaculture, 2007, 267(1 4): [8] Zehra S, Khan M A. Dietary phenylalanine requirement and tyrosine replacement value for phenylalanine for fingerling

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12 Dietary phenylalanine requirement of the GIFT strain of Nile tilapia, Oreochromis niloticus, reared in freshwater JIANG Ming 1, WU Wenyi 1, 2, WEN Hua 1, LIU Wei 1, WU Fan 1, TIAN Juan 1, YANG Changgeng 1 1. Key Laboratory of Freshwater Biodiversity Conservation and Utilization, Ministry of Agriculture; Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan , China; 2. College of Fisheries and Life Science, Shanghai Ocean University, Shanghai , China Abstract: Phenylalanine (Phe) is one of the essential amino acids for fish. It is an essential component of proteins that participate in many important physiological and metabolic processes, including growth and development. A deficiency or excess of phenylalanine in the diet can significantly affect the growth performance and/or feed utilization of farmed fish. Therefore, it is necessary to ensure that dietary levels of Phe meet, but do not exceed, the requirements of the fish. The GIFT strain of Oreochromis niloticus (Nile tilapia) is becoming an important and economically valuable farmed freshwater species. The aim of this study was to determine the optimal dietary Phe requirement of the GIFT strain by evaluating its growth performance, feed utilization, body composition, muscle amino acid composition, and digestive enzyme activities in a 60-day growth experiment. Triplicate groups of experimental tilapia (initial body weight, g) were cultured in freshwater in 15 floating net cages (1.0 m 1.0 m 1.5 m). Six isonitrogenous (30.10% crude protein) and isocaloric (17.73 MJ/kg gross energy) diets were formulated with protein sources including fish meal, gelatin, peanut meal, soybean meal, casein, and crystalline amino acids. The measured Phe levels in the experimental diets were 0.78%, 0.95%, 1.09%, 1.34%, 1.51%, and 1.72%. The dietary amino acid pattern, except for Phe, was adapted to the muscle amino acid pattern of the GIFT strain. The results showed that with increasing dietary Phe levels, the weight gain rate, specific growth rate, protein efficiency ratio, and protein deposition efficiency increased significantly (P<0.05) with the highest values in the 1.34% Phe group. The feed conversion ratio decreased significantly (P<0.05) with increasing dietary Phe levels, and showed the lowest value in the 1.09% Phe group. The hepatosomatic index, viscerosomatic index, and condition factor of the GIFT strain all increased significantly (P<0.05) as dietary Phe levels increased from 0.78% to 1.34%, but decreased when dietary Phe levels exceeded 1.34%. The highest whole body crude lipid and whole body ash contents were in the 1.34% dietary Phe group. There were no significant differences (P>0.05) among the six groups in whole body moisture content, whole body crude protein content, muscle moisture content, muscle crude protein content, muscle crude lipid content, and muscle amino acids contents. The muscle ash content of the group fed 0.98% dietary Phe was significantly (P<0.05) lower than that of the group fed 1.72% dietary Phe, but was not significantly different (P>0.05) from those of the other groups. The serum chemistry analysis showed that dietary Phe significantly affected the glucose, total cholesterol, triglyceride, and alanine aminotransferase contents (P<0.05), but not the aspartate aminotransferase content (P>0.05). The dietary Phe levels also significantly affected the activities of intestinal protease, intestinal lipase, and Na + -K + -ATPase (P<0.05), but not intestinal amylase activity. The quadratic regression analyses of weight gain rate, feed conversion ratio, and protein efficiency ratio showed that the optimum dietary Phe requirement of GIFT is 1.17% 1.21% of the diet or 3.89% 4.02% of dietary proteins. Key words: GIFT(Oreochromis niloticus); phenylalanine; dietary requirement; growth Corresponding author: WEN Hua. wenhua.hb@163.com

中草药 Chinese Traditional and Herbal Drugs 第 46 卷第 9 期 2015 年 5 月 1339 非酒精性脂肪性肝病 (nonalcoholic fatty liver disease,nafld) 是一种与胰岛素抵抗遗

中草药 Chinese Traditional and Herbal Drugs 第 46 卷第 9 期 2015 年 5 月 1339 非酒精性脂肪性肝病 (nonalcoholic fatty liver disease,nafld) 是一种与胰岛素抵抗遗 1338 中草药 Chinese Traditional and Herbal Drugs 第 46 卷第 9 期 2015 年 5 月九味肝泰胶囊对高脂饮食诱导大鼠非酒精性脂肪肝的治疗作用陈菲 1 2,, 艾国 3, 盛柳青 1, 麻佳蕾 1, 黄正明 1. 金华职业技术学院医学院, 浙江金华