Effect of dietary guanidinoacetic acid on growth, hepatic anti-oxidative capacity and muscle energy metabolism in channel catfish (Ictalurus punctatus)
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摘要: 为研究饲料中添加胍基乙酸（GAA）对斑点叉尾鮰生长、肝脏抗氧化以及肌肉能量代谢的影响。本实验共设计4个实验组，分别投喂含0（对照组）、300、600和900 mg/kg GAA（GAA0、GAA300、GAA600和GAA900）的等能等氮饲料，实验周期8周。结果显示，各添加GAA组末均重（FBW）和增重率（WGR）较对照组均有提高趋势，但差异不显著；添加GAA显著影响脏体比（VSI）和腹脂率（IPF），在GAA300组VSI和IPF最低；以IPF和VSI为评价指标，通过二次函数分析获得GAA最适量分别为275和150 mg/kg。添加GAA对全鱼水分、粗蛋白、粗脂肪和灰分均无显著影响，对肌肉中游离氨基酸和脂肪酸均无显著影响，但能显著提高肌肉中羟脯氨酸（HYP）含量，改善肌肉品质。添加GAA可提高肝脏抗氧化能力，其中GAA300组的总抗氧化能力（T-AOC）和超氧化物歧化酶（SOD）活性显著高于对照组。添加GAA可改善肌肉能量代谢，其中GAA300组的琥珀酸脱氢酶（SDH）和丙酮酸激酶（PK）活性显著高于对照组。研究表明，饲料中添加GAA对斑点叉尾鮰生长及饲料利用无显著促进作用，但添加150~300 mg/kg GAA能降低斑点叉尾鮰腹部脂肪含量和脏体比，并能提高肝脏抗氧化能力和肌肉能量代谢。Abstract: A 8-week feeding trial was conducted to study the effect of dietary guanidinoacetic acid (GAA) on the growth performance, hepatic anti-oxidative capacity and muscle energy metabolism of channel catfish (Ictalurus punctatus). Four isonitrogenous and isocaloric experimental diets with 0 (control diet), 300, 600, and 900 mg/kg GAA (GAA0, GAA300, GAA600, and GAA900) were formulated. The results showed that appropriate dietary GAA could improve the average final body weight (FBW) and weight gain rate (WGR), but not significantly. While the viscerosomatic index (VSI) and intraperitoneal fat ratio (IPF) of experiemental I. punctatus were significantly affected by dietary GAA levels, with the lowest values in GAA300 group. Based on the quadratic regression analysis of IFP and VSI, the optimal dietary GAA level for I. punctatus was 275 mg/kg and 150 mg/kg, respectively. The addition of GAA in diet had no significant effect on moisture content, crude protein, crude fat and ash content of whole fish, as well as the free amino acids and fatty acids in muscle. While the hydroxyproline (HYP) in muscle was significantly increased with the increase of dietary GAA for better muscle quality. Additionally, dietary GAA can improve hepatic anti-oxidative capacity. The total antioxidative capacity (T-AOC) and superoxide dismutases (SOD) activities of fish in GAA300 group were significantly higher than those of the control group. GAA can also improve muscle energy metabolism, and the succinate dehydrogenase (SDH) and pyruvate kinase (PK) activities of fish in GAA300 group were significantly higher than those of the control group. Conclusively, dietary GAA has no significant effect on growth performance and feed utilization of I. punctatus. While 150-300 mg/kg GAA in diet can reduce intraperitoneal fat content and viscerosomatic index, and improve hepatic anti-oxidative capacity and muscle energy metabolism of I. punctatus.
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图 1 饲料中GAA水平对斑点叉尾鮰腹脂率的影响
Figure 1. Effect of different dietary GAA supplementation on IPF of I. punctatus
图 2 饲料中GAA水平对斑点叉尾鮰脏体比的影响
Figure 2. Effect of different dietary GAA supplementations on VSI of I. punctatus
表 1 基础饲料配方及常规成分(干物质)
Table 1. Formulation and proximate composition of the basal diet (air-dry basis)
% 项目 items 含量 content 项目 items 含量 content 国产鱼粉(62%) domestic fish meal 5.00 蛋氨酸 methionine 0.20 猪肉粉(69%) pork meat powder 8.00 盐 NaCl 0.50 豆粕(46%) soybean meal 34.00 矿物元素预混料1 mineral premix 0.50 脱酚棉籽蛋白(60%) degossypolized cottonseed protein 5.00 维生素预混料2 vitamin premix 0.20 菜粕(36%) rapeseed meal 10.00 氯化胆碱(75%) choline chloride 0.25 面粉(15%) wheat flour 20.00 维生素C(35%) vitamin C 0.05 米糠(13%) rice bran 9.38 抗氧化剂 antioxidants 0.02 鱼油 fish oil 1.00 合计 total 100.00 豆油 soybean oil 3.00 营养水平 nutrient levels 磷酸二氢钙 calcium diphosphate 2.00 粗蛋白 crude protein 37.48 酿酒酵母 brewer,s yeast meal 0.50 粗脂肪 crude lipid 6.67 赖氨酸 lysine 0.40 粗灰分 crude ash 8.05 注：1. 每千克维生素元素预混料含有维生素A乙酸酯480 mg、维生素D3 7.5 mg、维生素E 10 668 mg、维生素K3 800 mg、维生素B2 1 500 mg、维生素B6 1 600 mg、烟酸7 800 mg、泛酸钙4 700 mg、叶酸1 100 mg、水分12%；2. 每千克矿物元素预混料含锰2 000 mg、铁90 000 mg、锌5 000 mg、铜300 mg、镁47 000 mg、硒10 mg、水分12%。 Notes: One kilogram of vitamin premix contains VA acetate 480 mg, VD3 7.5 mg, VE 10 668 mg, VK3 800 mg, VB2 1 500 mg, VB6 1 600 mg, niacin 7 800 mg, calcium pantothenate 4 700 mg, folic acid 1 100 mg, moisture 12%; one kilogram of mineral premix contained Mn 2 000 mg, Fe 90 000 mg, Zn 5 000 mg, Gu 300 mg, Mg 47 000 mg, Se 10 mg, moisture 12%. 
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表 2 饲料中GAA水平对斑点叉尾鮰生长和饲料利用的影响
Table 2. Effect of different dietary GAA supplementation on growth performance and feed utilization of I. punctatus
项目 items GAA0 GAA300 GAA600 GAA900 P值 P-value 初始体重/g IBW 36.67±0.00 37.09±0.27 36.88±0.21 36.91±0.51 0.815 终末体重/g FBW 145.96±0.89 147.37±0.99 147.22±1.13 146.88±0.51 0.780 增重率/% WGR 298.06±2.43 299.24±3.74 299.24±3.03 298.55±2.32 0.989 特定生长率/% SGR 2.47±0.01 2.47±0.02 2.47±0.01 2.47±0.01 0.983 饲料系数/% FCR 1.18±0.01 1.18±0.01 1.18±0.02 1.19±0.01 0.964 肝体比/% HSI 1.13±0.04 1.13±0.04 1.17±0.04 1.18±0.01 0.633 脏体比/% VSI 7.74±0.16ab 7.51±0.18a 8.06±0.22ab 8.43±0.06b 0.014 腹脂率/% IPF 3.94±0.24ab 3.72±0.15a 4.06±0.05ab 4.58±0.25b 0.043 肥满度/(g/cm3) CF 1.54±0.03 1.59±0.05 1.54±0.03 1.53±0.04 0.748 成活率/% SR 100.00±0.00 99.45±0.56 99.45±0.56 98.89±1.11 0.724 注：同一行数值上标不同字母表示差异显著(P < 0.05)；下同。
Notes: Values in the same row with different superscripts are significant different (P < 0.05); the same below.
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表 3 饲料中GAA水平对斑点叉尾鮰体成分的影响
Table 3. Effect of different dietary GAA supplementation on body nutritional composition of I. punctatus
项目 items GAA0 GAA300 GAA600 GAA900 P值 P-value 水分/% moisture 71.87±0.53 72.07±0.05 71.87±0.40 72.00±0.48 0.980 粗蛋白/% crude protein 14.07±0.42 14.28±0.31 14.07±0.07 14.06±0.18 0.929 粗脂肪/% crude lipid 9.67±0.26 9.13±0.34 9.77±0.32 9.11±0.28 0.326 粗灰分/% crude ash 3.11±0.07 3.26±0.09 3.19±0.04 3.17±0.01 0.432
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表 4 饲料中GAA水平对斑点叉尾鮰肌肉糖原和羟脯氨酸含量的影响
Table 4. Effect of different dietary GAA supplementation on glycogen and hydroxyproline in muscle of I. punctatus
项目 items GAA0 GAA300 GAA600 GAA900 P值 P-value 肌糖原/(mg/g) muscle glycogen 0.38±0.02 0.40±0.02 0.42±0.02 0.43±0.02 0.298 羟脯氨酸/(μg/g) HYP 8.40±1.01a 14.36±0.81b 20.83±0.79c 25.94±1.51c 0.000
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表 5 饲料中GAA水平对斑点叉尾鮰肌肉氨基酸组成的影响
Table 5. Effect of different dietary GAA supplementation on free amino acid composition in muscle of I. punctatus (dry matter) μg/g
项目 items GAA0 GAA300 GAA600 GAA900 P值 P-value 苏氨酸 Thr 184.33±10.01 201.73±14.34 189.11±10.49 172.62±4.86 0.297 缬氨酸 Val 60.01±2.75 57.02±2.30 57.07±3.32 58.08±5.58 0.179 蛋氨酸 Met 2.48±0.35 1.92±0.27 1.60±0.44 1.80±0.23 0.290 异亮氨酸 Ile 18.78±3.88 21.34±2.53 19.71±2.65 28.58±2.03 0.104 亮氨酸 Leu 60.33±4.00 64.65±1.58 68.60±3.74 61.05±3.01 0.276 苯丙氨酸 Phe 172.36±7.54 152.62±15.41 144.59±6.07 141.17±14.99 0.271 赖氨酸 Lys 159.91±12.28 147.81±14.22 184.69±15.32 158.96±14.94 0.337 组氨酸 His 51.93±3.80 53.97±2.08 59.61±5.26 54.96±3.55 0.605 精氨酸 Arg 77.10±3.09 70.61±9.45 67.36±10.42 67.29±6.24 0.770 脯氨酸 Pro 71.20±4.78 68.97±4.98 69.78±2.28 78.78±7.42 0.622 天冬氨酸 Asp 14.86±1.37 16.22±0.90 16.47±2.88 17.14±2.32 0.821 丝氨酸 Ser 61.89±5.30 55.43±2.77 55.80±5.43 64.22±6.50 0.569 谷氨酸 Glu 157.21±7.20 157.73±4.77 154.04±7.96 147.49±17.13 0.865 甘氨酸 Gly 222.14±19.53 207.06±15.73 186.84±15.58 187.31±12.73 0.363 丙氨酸 Ala 302.87±3.50 323.85±13.27 325.52±10.07 315.02±15.24 0.559 半胱氨酸 Cys 18.43±1.73 18.64±1.01 16.88±0.29 18.61±2.86 0.868 酪氨酸 Tyr 66.60±3.17 76.33±1.67 70.09±1.63 73.39±4.01 0.115 非必需氨基酸 NEAA 1013.18±14.53 1046.97±48.55 1009.20±38.29 1002.79±58.91 0.896 必需氨基酸 EAA 1711.26±59.96 1697.04±77.19 1701.59±23.15 1648.52±87.57 0.923
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表 6 饲料中GAA水平对斑点叉尾鮰肌肉脂肪酸含量的影响
Table 6. Effect of different dietary GAA supplementation on the fatty acid (FA) of total FA in muscle of I. punctatus
% 项目 items GAA0 GAA300 GAA600 GAA900 P值 P-value C14:0 1.06±0.02 1.13±0.04 1.15±0.06 1.07±0.02 0.299 C15:0 0.17±0.00 0.17±0.00 0.17±0.00 0.17±0.00 0.863 C16:0 19.61±0.12 19.20±0.25 19.82±0.26 19.71±0.31 0.337 C17:0 0.25±0.01 0.25±0.01 0.24±0.01 0.25±0.01 0.674 C18:0 8.73±0.10 8.50±0.33 7.97±0.32 8.56±0.14 0.191 C20:0 0.25±0.01 0.27±0.00 0.25±0.01 0.26±0.00 0.357 C22:0 0.07±0.00 0.08±0.00 0.08±0.01 0.08±0.00 0.427 C24:0 1.45±0.08 1.26±0.08 1.34±0.12 1.46±0.09 0.409 C16:1 1.41±0.02 1.48±0.07 1.56±0.09 1.47±0.03 0.414 C18:1n9t 0.22±0.00 0.22±0.01 0.21±0.01 0.20±0.01 0.206 C18:1n9c 35.40±0.99 36.29±0.52 36.23±0.77 35.19±0.99 0.718 C20:1 1.25±0.03 1.32±0.03 1.24±0.04 1.21±0.01 0.063 C18:2n6c 18.06±0.23 19.29±0.28 18.30±0.54 18.24±0.13 0.088 C18:3n6 0.22±0.01 0.25±0.01 0.25±0.02 0.23±0.01 0.193 C18:3n3 1.50±0.04 1.66±0.03 1.54±0.08 1.53±0.04 0.154 C20:2 1.31±0.05 1.18±0.03 1.18±0.04 1.26±0.00 0.115 C20:3n3 0.12±0.01 0.11±0.01 0.11±0.01 0.12±0.00 0.465 C20:3n6 1.55±0.10 1.39±0.06 1.34±0.05 1.59±0.11 0.159 C20:4n6 1.68±0.12 1.25±0.12 1.33±0.13 1.44±0.10 0.079 C20:5n3 (EPA) 0.02±0.01 0.02±0.01 0.02±0.01 0.02±0.01 0.979 C22:6n3 (DHA) 5.74±0.49 4.52±0.32 4.50±0.26 5.22±0.28 0.072 n3 PUFA 7.57±0.40 6.32±0.30 6.89±0.65 7.26±0.40 0.546 n6 PUFA 21.51±0.24 22.17±0.27 21.55±0.28 21.80±0.30 0.301 n3/n6 0.35±0.02 0.29±0.01 0.31±0.03 0.33±0.02 0.324 饱和脂肪酸 SFA 31.29±0.44 30.88±0.19 31.06±0.40 31.06±0.44 0.595 单不饱和脂肪酸 MUFA 38.30±1.03 39.38±0.58 39.30±0.89 38.07±1.07 0.659 多不饱和脂肪酸 PUFA 30.96±0.32 29.74±0.44 29.63±0.54 30.33±0.68 0.332
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表 7 饲料中GAA水平对斑点叉尾鮰肝脏抗氧化指标的影响
Table 7. Effect of different dietary GAA supplementation on hepatic antioxidant indices of I. punctatus
项目 items GAA0 GAA300 GAA600 GAA900 P值 P-value 总抗氧化力/(U/mg prot) T-AOC 0.12±0.01a 0.17±0.01b 0.13±0.00a 0.12±0.01a 0.003 谷胱甘肽过氧化物酶/(U/mg prot) GSH-Px 20.91±3.89 26.22±6.70 23.50±1.36 24.03±0.49 0.820 丙二醛/(nmol/mg prot) MDA 0.41±0.02a 0.62±0.10a 1.10±0.12b 1.12±0.12b 0.001 超氧化物歧化酶/(U/mg prot) SOD 966.18±6.81a 1191.42±26.06b 1236.50±65.17b 1133.99±40.61ab 0.007 过氧化氢酶/(U/mg prot) CAT 0.43±0.02 0.43±0.03 0.45±0.05 0.45±0.04 0.944
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表 8 饲料中GAA水平对斑点叉尾鮰肌肉能量代谢酶的影响
Table 8. Effect of different dietary GAA supplementation on the activities of muscle energy enzymes of I. punctatus
项目 items GAA0 GAA300 GAA600 GAA900 P值 P-value 肌酸激酶/(U/mg prot) CK 5.74±0.23 5.84±0.25 6.13±0.19 6.03±0.23 0.668 琥珀酸脱氢酶/(U/mg prot) SDH 11.11±0.11a 14.02±0.66b 12.88±0.70ab 13.31±0.54ab 0.028 丙酮酸激酶/(U/g prot) PK 82.59±1.44a 96.00±2.45b 103.98±2.73b 84.85±2.40a 0.001
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