Chinese Journal of Animal Nutrition >

2025 , Vol. 37 >Issue 3: 1929 - 1939

DOI: https://doi.org/10.12418/CJAN2025.163

RESEARCH PAPER

Effects of Dietary Tryptophan and Methionine on Growth Performance, Digestive Ability, Immune Function and Antioxidant Capacity of Chinese Mitten Crab (Eriocheir sinensis)

  • JIANG Xianghui ,
  • WANG Xingbing ,
  • LI He ,
  • HU Zongyun ,
  • ZHANG Jian ,
  • WEI Hongxiang ,
  • YANG Peimin , *
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  • Liaoning Key Laboratory of Aquatic Animal Disease Control, Liaoning Institute of Freshwater Fisheries, Liaoyang 111000, China
*professor, E-mail:

Received date: 2024-09-03

  Online published: 2025-03-13

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Abstract

This experiment was conducted to investigate the effects of dietary tryptophan and methionine on growth performance, digestive ability, immune function and antioxidant capacity of Chinese mitten crab (Eriocheir sinensis). In a two-factor experimental design, 0, 0.31% and 0.63% L-Try and 0, 0.33% and 0.67% L-Met were added to the basal diet, nine groups of isonitrogen and isolipid diets were prepared with three Try levels (actual contents were 0.31%, 0.62% and 0.94%) and three Met levels (actual contents were 0.66%, 0.99% and 1.33%), and each group contained 3 replicates. The Eriocheir sinensis with initial body weight of (50.27±2.20) g were selected, and the stocking density was 4 500 crab/hm2. The pre-experimental period lasted for 10 days, and the experimental period lasted for 80 days. The results showed as follows: 1) dietary Try and Met levels had not significant interaction effect on the growth performance of Eriocheir sinensis (P>0.05). The weight gain rate, specific growth rate and protein efficiency of 0.94% Try level and 0.99% Met level were significantly higher than those of other levels (P<0.05), and the feed conversion rate was significantly lower than that of other levels (P<0.05). 2) Dietary Try and Met levels had significant interaction effects on muscle crude protein and rude ash content of Eriocheir sinensis (P<0.05). The muscle crude protein content of 0.94% Try level and 0.99% and 1.33% Met level was significantly higher than that of other levels (P<0.05), and the muscle crude ash content was significantly lower than that of other levels (P<0.05). 3) Dietary Try and Met levels had significant interaction effects on hepatopancreas pepsin, trypsin, lipase and amylase activities of Eriocheir sinensis (P<0.05). The activities of pepsin and trypsin in hepatopancreas of 0.94% Try level and 0.99% Met level were significantly higher than those of other levels (P<0.05), the activities of lipase and amylase in hepatopancreas of 0.62%Try level and 0.99% Met level were significantly higher than those of other levels (P<0.05). 4) Dietary Try and Met levels had significant interaction effects on the serum non-specific immune (except acid phosphatase) and antioxidant indexes of Eriocheir sinensis (P<0.05). In the non-specific immune indexes, the activities of alkaline phosphatase and lysozyme in serum of 0.94% Try level and 0.99% Met level were significantly higher than those of other levels (P<0.05). In the antioxidant indexes, the activities of catalase and superoxide dismutase in serum of 0.94% Try level were significantly higher than those of other levels (P<0.05), and the malondialdehyde content was significantly lower than that of other levels (P<0.05); the activities of catalase, superoxide dismutase and total antioxidant capacity in serum of 0.99% Met level were significantly higher than those of other levels (P<0.05). In conclusion, the appropriate dietary Try and Met can improve the growth performance of Eriocheir sinensis, and the Try and Met supplemental levels and their interaction affect the muscle crude protein content, hepatopancreas digestive enzyme activity, non-specific immunity and antioxidant capacity; comprehensive all indexes, the best results are obtained when the levels of Try and Met in the diet are 0.94% and 0.99%, respectively.

Key words： Eriocheir sinensis; tryptophan; methionine; growth; body composition; digestive enzymes; immune function; antioxidant capacity

Cite this article

JIANG Xianghui , WANG Xingbing , LI He , HU Zongyun , ZHANG Jian , WEI Hongxiang , YANG Peimin . Effects of Dietary Tryptophan and Methionine on Growth Performance, Digestive Ability, Immune Function and Antioxidant Capacity of Chinese Mitten Crab (Eriocheir sinensis)[J]. Chinese Journal of Animal Nutrition, 2025 , 37(3) : 1929 -1939 . DOI: 10.12418/CJAN2025.163

中华绒螯蟹(Eriocheir sinensis)又名河蟹、毛蟹、大闸蟹,具有重要的营养价值和经济价值,是我国主要养殖蟹类之一[1]。色氨酸(Try)和蛋氨酸(Met)是水产动物体内不能合成的必需氨基酸,是中华绒螯蟹生长的第一、第二限制性氨基酸[2⇓-4]。到目前为止,对于中华绒螯蟹氨基酸营养需求量的研究仅局限于单个氨基酸需求量[5⇓-7],并没有针对相关性较强的氨基酸之间的交互作用进行研究。近些年来,越来越多的学者把饲料中氨基酸交互作用作为水产动物营养生理研究的重点,研究对象主要集中在鱼类和棘皮动物[8⇓-10]。Try和Met同属于中性氨基酸,具有功能相关性,Try除了有重要的生物学功能,还能节约Met,它可以通过代谢途径转化为5-羟色氨酸和色酮酸,从而代替Met参与蛋白质合成过程[11]。养殖过程中,有效利用Try和Met,不仅有利于提高动物的生长性能和健康水平,还可提高动物的品质、缓解蛋白质饲料资源紧缺和养殖废弃物造成的环境污染等问题。因此,本试验旨在探究饲料中添加Try和Met对中华绒螯蟹生长性能、体成分、消化能力、免疫功能及抗氧化能力的影响,以确定中华绒螯蟹饲粮中Try和Met的适宜添加水平。

1 材料与方法

1.1 试验设计和饲料

基础饲料主要蛋白质来源为鱼粉、虾粉、豆粕、菜籽粕,主要脂类来源为大豆油。在满足中华绒螯蟹饲料需求的基础上,采用3×3双因素试验设计了9组(1~9组)等氮(42%)等脂(6%)的饲料[5⇓-7],基础饲料中分别添加0、0.31%、0.63%的L-Try和0、0.33%、0.67%的L-Met,配制成3个Try水平(实际含量分别为0.31%、0.62%、0.94%)×3个Met水平(实际含量分别为0.66%、0.99%、1.33%)的试验饲料。试验饲料组成及营养水平见表1。饲料原料经粉碎过60目筛,混合均匀后用制粒机制成粒径为2 mm的颗粒饲料,55 ℃烘干24 h自然冷却后放入塑料自封袋中,于-20 ℃冰箱中保存备用。试验用晶体L-Try和L-Met实测含量>99.7%。
表1 试验饲料组成及营养水平(风干基础)

Table 1 Composition and nutrient levels of experimental diets (air-dry basis) %

项目
Items		 组别Groups
1 2 3 4 5 6 7 8 9
原料Ingredients
鱼粉Fish meal 21.86 21.86 21.86 21.86 21.86 21.86 21.86 21.86 21.86
虾粉Shrimp powder 8.72 8.72 8.72 8.72 8.72 8.72 8.72 8.72 8.72
豆粕Soybean meal 30.05 30.05 30.05 30.05 30.05 30.05 30.05 30.05 30.05
菜籽粕Rapeseed meal 13.95 13.95 13.95 13.95 13.95 13.95 13.95 13.95 13.95
面粉Wheat flour 11.28 11.28 11.28 11.28 11.28 11.28 11.28 11.28 11.28
麦麸Wheat bran 8.02 7.71 7.39 7.69 7.38 7.06 7.35 7.04 6.72
大豆油Soybean oil 2.12 2.12 2.12 2.12 2.12 2.12 2.12 2.12 2.12
磷酸二氢钙Ca(H2PO4)2 2.50 2.50 2.50 2.50 2.50 2.50 2.50 2.50 2.50
预混料Premix1) 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00
食盐NaCl 0.50 0.50 0.50 0.50 0.50 0.50 0.50 0.50 0.50
L-色氨酸L-Try 0.31 0.63 0.31 0.63 0.31 0.63
L-蛋氨酸L-Met 0.33 0.33 0.33 0.67 0.67 0.67
合计Total 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00 100.00
营养水平Nutrient levels2)
总能GE/(MJ/kg) 18.25 18.28 18.26 18.24 18.24 18.28 18.26 18.24 18.24
粗蛋白质CP 42.22 41.97 42.23 41.99 42.13 42.07 42.23 42.04 42.11
粗脂肪EE 6.17 6.15 6.18 6.25 6.17 6.19 6.18 6.25 6.19
水分Moisture 10.11 10.15 10.04 10.19 10.01 10.15 10.04 10.19 10.01
粗灰分Ash 4.78 4.75 4.84 4.80 4.77 4.75 4.84 4.80 4.77
色氨酸Try 0.31 0.62 0.93 0.32 0.61 0.94 0.30 0.62 0.94
蛋氨酸Met 0.66 0.65 0.66 0.98 0.99 0.99 1.32 1.33 1.33

1)预混料为每千克饲料提供 The premix provided the following per kg of diets: Cu 20 mg,Fe 25 mg,Mg 12 mg,Mn 26 mg,Zn 80 mg,Se 0.16 mg,I 0.45 mg,Co 0.6 mg,VA 10 000 IU,VE 100 mg,VC 100 mg,VD 32 500 IU,VK 32.2 mg,VB1 3.2 mg,VB2 15 mg,VB3 20 mg,VB4 10 mg,VB5 25 mg,VB12 0.016 mg,叶酸 folic acid 5 mg,胆碱 choline 600 mg,生物素 biotin 0.15 mg,肌醇 inositol 200 mg。

2)营养水平为实测值。Nutrient levels were measured values.

1.2 饲养试验和管理

试验方案经辽宁省淡水水产科学院学术委员会批准(批准号:2023014)。在辽宁省农科院所属稻鱼蟹健康生态研发基地(盘锦绕阳农业科技发展有限公司)选取9块平均面积为1.04 hm2的相邻稻田,用养殖专用的聚乙烯塑料薄膜平均分成3块,四角围成半圆弧型,以防止逃逸和敌害。2023年5月底至6月初进行水稻插秧,选择的水稻品种为盐粳939号,株行距为30 cm×18 cm,水深15~18 cm。在插秧结束1个月后投放附肢完整、个体均匀的健康中华绒螯蟹,平均体重为(50.27±2.20) g/尾,密度为4 500只/hm2,每天15:00按蟹体重的3%~5%在固定点投喂蟹料,固定点设置在靠近岸边的位置。每隔20 d监测1次水质指标和水中浮游动植物情况,避免蟹摄食昆虫及水生动植物对试验产生影响。试验预试期10 d,正试期80 d。

1.3 样品采集和指标测定

试验结束后干塘并全部过数,测量体重并记录存活只数。每组随机抽取30只雌蟹和30只雄蟹,擦干体表水分并用1.0 mL无菌注射器从步足基部抽取血淋巴,静置4 h后,在4 ℃下离心10 min,取血清装入冻存管,置于-80 ℃冰箱保存,用于血清免疫和抗氧化指标测定。解剖取出肝胰腺,-40 ℃保存待测。
饲料总能采用C6000氧弹量热仪(ISO 9831—1998.08.01)测定,饲料和蟹的水分、粗蛋白质、粗脂肪和粗灰分含量分别采用105 ℃恒重法(GB/T 6435—2014)、凯氏定氮法(GB/T 6432—2018)、索氏抽提法(GB/T 6433—2006)和550 ℃马福炉灼烧法(GB/T 6438—2007)测定[12]。
采用试剂盒(南京建成生物工程研究所)测定肝胰腺中消化酶[胃蛋白酶(pepsin)、胰蛋白酶(trypsin)、脂肪酶(lipase)和淀粉酶(amylase)]活性,血清非特异性免疫指标[碱性磷酸酶(AKP)、酸性磷酸酶(ACP)、溶菌酶(LZM)和酚氧化酶(PO)活性]和抗氧化相关指标[超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性和总抗氧化能力(T-AOC)及丙二醛(MDA)含量]。

1.4 计算公式

生长性能相关指标计算公式如下:
增重率(WGR,%)=[(Wt-W0)/W0]×100;
饵料系数(FCR)=Wf/(Wt-W0);
特定生长率(SGR,%/d)=[(lnWt-lnW0)/t]×100;
蛋白质效率(PER,%)=[(Wt-W0)/(Wf×饲料粗蛋白质含量)]×100。
式中:W0为试验蟹初始均重;Wt为试验蟹终末均重;t为饲喂天数;Wf为试验蟹摄食饲料重(风干基础)。

1.5 统计分析

试验数据先用Excel 2019初步处理后,用SPSS 20.0统计软件进行双因素方差分析(two-way ANOVA)。各组之间的差异显著性用Duncan氏多重比较法分析,P<0.05表示差异显著,数据以平均值±标准差(mean±SD)表示。

2 结果

2.1 饲料中添加Try和Met对中华绒螯蟹生长性能的影响

由表2可知,饲料Try和Met水平对中华绒螯蟹生长性能无显著交互作用(P>0.05)。饲料Try和Met水平对中华绒螯蟹增重率、饵料系数、特定生长率和蛋白质效率均产生显著影响(P<0.05)。随着饲料中Try水平的提升,增重率、特定生长率和蛋白质效率呈现上升趋势,饵料系数呈现下降趋势,Try水平为0.94%时效果最好;随着饲料中Met水平的提升,增重率、特定生长率和蛋白质效率则呈现先升高后降低的趋势,饵料系数呈现先降低后升高的趋势,Met水平为0.99%时效果最好。
表2 饲料中添加色氨酸和蛋氨酸对中华绒螯蟹生长性能的影响

Table 2 Effects of dietary Try and Met on growth performance of Eriocheir sinensis

项目
Items		 增重率
WGR/%		 饵料系数
FCR		 特定生长率
SGR/(%/d)		 蛋白质效率
PER/%
组别Groups
1 182.96±6.39a 3.25±0.11c 1.30±0.05a 0.73±0.03a
2 202.86±5.14b 2.93±0.07bc 1.39±0.04ab 0.82±0.02ab
3 210.25±7.88b 2.83±0.10b 1.42±0.08ab 0.85±0.04ab
4 226.76±3.69b 2.62±0.08b 1.48±0.09b 0.91±0.01b
5 222.87±5.48b 2.67±0.07b 1.47±0.11b 0.90±0.02ab
6 288.30±5.56c 2.10±0.13a 1.69±0.06c 1.16±0.08c
7 213.93±6.04b 2.78±0.06b 1.43±0.13ab 0.86±0.02ab
8 207.74±5.22b 2.86±0.09b 1.41±0.07ab 0.84±0.04ab
9 218.99±4.17b 2.72±0.20b 1.45±0.06b 0.88±0.03ab
色氨酸水平Try levels/%
0.31 207.88±7.43a 2.88±0.10b 1.40±0.04a 0.83±0.03a
0.62 211.15±7.43a 2.82±0.10b 1.42±0.04a 0.85±0.03a
0.94 239.18±7.43b 2.55±0.10a 1.52±0.04b 0.96±0.03b
蛋氨酸水平Met levels/%
0.66 198.69±7.43a 3.00±0.10c 1.37±0.04a 0.80±0.03a
0.99 245.97±7.43b 2.46±0.10a 1.55±0.04b 0.99±0.03b
1.33 213.55±7.43a 2.79±0.10b 1.42±0.04a 0.86±0.03a
双因素方差分析Two-way ANOVA
色氨酸水平Try level * * * *
蛋氨酸水平Met level * * * *
交互Interaction ns ns ns ns

*表示显著(P<0.05),ns表示不显著(P>0.05)。同列数据肩标不同小写字母表示差异显著(P<0.05),相同或无字母表示差异不显著(P>0.05)。下表同。

* indicated significant (P<0.05), ns indicated no significant (P>0.05). In the same column, values with different small letter superscripts mean significant difference (P<0.05), while with the same or no letter superscripts mean no significant difference (P>0.05). The same as below.

2.2 饲料中添加Try和Met对中华绒螯蟹体成分的影响

由表3可知,饲料Try和Met水平对中华绒螯蟹肌肉中粗蛋白质和粗灰分含量均有显著交互作用(P<0.05)。在饲料Try水平中,肌肉中粗蛋白质含量的最低值与最高值分别出现在0.31%和0.94% Try水平,且3个水平之间差异显著(P<0.05);肌肉中粗灰分含量的最低值与最高值分别出现在0.94%和0.31% Try水平,且3个水平之间差异显著(P<0.05)。在饲料Met水平中,0.99%和1.33% Met水平的肌肉中粗蛋白质含量最高,且显著高于0.66% Met水平(P<0.05);0.66 % Met水平的肌肉中粗灰分含量最高,且显著高于0.99%和1.33% Met水平(P<0.05)。
表3 饲料中添加色氨酸和蛋氨酸对中华绒螯蟹体成分的影响(鲜重基础)

Table 3 Effects of dietary Try and Met on body composition of Eriocheir sinensis (fresh weight basis) %

项目Items 水分Moisture 粗蛋白质CP 粗脂肪EE 粗灰分Ash
色氨酸水平Try levels/%
0.31 77.40±0.87 16.65±0.38a 0.85±0.03 2.76±0.07c
0.62 77.34±0.87 16.93±0.38b 0.86±0.03 2.61±0.07b
0.94 77.54±0.87 17.97±0.38c 0.86±0.03 2.43±0.07a
蛋氨酸水平Met levels/%
0.66 77.49±0.87 16.97±0.38a 0.85±0.03 2.71±0.07b
0.99 77.55±0.87 17.32±0.38b 0.85±0.03 2.57±0.07a
1.33 77.23±0.87 17.35±0.38b 0.86±0.03 2.53±0.07a
双因素方差分析Two-way ANOVA
色氨酸水平Try level ns * ns *
蛋氨酸水平Met level ns * ns *
交互Interaction ns * ns *

2.3 饲料中添加Try和Met对中华绒螯蟹消化酶活性的影响

由表4可知,饲料Try和Met水平对中华绒螯蟹肝胰腺中胃蛋白酶、胰蛋白酶、脂肪酶和淀粉酶活性均有显著交互作用(P<0.05)。肝胰腺中胃蛋白酶和胰蛋白酶活性随着饲料Try水平的上升而升高,随着Met水平的上升呈现先升高后降低的趋势,其中0.94% Try水平和0.99% Met水平的肝胰腺中胃蛋白酶和胰蛋白酶活性显著高于其他水平(P<0.05)。肝胰腺中脂肪酶和淀粉酶活性随着饲料Try和Met添加水平的上升呈现先升高后降低的趋势,其中0.62% Try水平和0.99% Met水平的肝胰腺中脂肪酶和淀粉酶活性显著高于其他水平(P<0.05)。
表4 饲料中添加色氨酸和蛋氨酸对中华绒螯蟹肝胰腺中消化酶活性的影响

Table 4 Effects of dietary Try and Met on hepatopancreas digestive enzyme activities of Eriocheir sinensis

项目
Items		 胃蛋白酶
Pepsin/
(U/mg prot)		 胰蛋白酶
Trypsin/
(U/mg prot)		 脂肪酶
Lipase/
(U/g prot)		 淀粉酶
Amylase/
(U/mg prot)
色氨酸水平Try levels/%
0.31 0.69±0.13a 5 537.60±769.24a 1.09±0.03a 2.89±0.11a
0.62 0.75±0.13b 8 954.42±769.24b 1.24±0.03b 4.19±0.11b
0.94 0.88±0.13c 9 977.27±769.24c 1.02±0.03a 2.87±0.11a
蛋氨酸水平Met levels/%
0.66 0.60±0.13a 6 548.07±769.24a 0.95±0.03a 3.22±0.11a
0.99 1.07±0.13b 10 048.60±769.24c 1.25±0.03c 3.83±0.11b
1.33 0.63±0.13a 8 835.65±769.24b 1.10±0.03b 3.23±0.11a
双因素方差分析Two-way ANOVA
色氨酸水平Try level * * * *
蛋氨酸水平Met level * * * *
交互Interaction * * * *

2.4 饲料中添加Try和Met对中华绒螯蟹血清非特异性免疫力及抗氧化性能的影响

由表5可知,除了血清ACP活性,饲料Try和Met水平对中华绒螯蟹血清其他免疫指标和抗氧化指标均有显著交互作用(P<0.05)。血清非特异性免疫指标中,AKP、LZM和PO活性随着饲料Try水平的上升而升高,随着饲料Met水平的上升呈先升高后降低的趋势。血清抗氧化指标中,CAT和SOD活性随着饲料Try水平的上升而升高,T-AOC则呈先升高后降低的趋势,MDA活性随着饲料Try水平的上升而下降;随着饲料Met水平的上升,CAT、SOD和T-AOC活性均呈先升高后降低的趋势,且0.99% Met水平显著高于其他水平(P<0.05)。
表5 饲料中添加色氨酸和蛋氨酸对中华绒螯蟹血清抗氧化和非特异性免疫指标的影响

Table 5 Effects of dietary Try and Met on serum antioxidant and non-specific immune indexes of Eriocheir sinensis

项目
Items		 碱性
磷酸酶
AKP/(金氏
单位/mL)		 酸性
磷酸酶
ACP/
(U/mL)		 溶菌酶
LZM/
(U/mL)		 酚氧化酶
PO/
(U/mL)		 过氧
化氢酶
CAT/
(U/mL)		 超氧化物
歧化酶
SOD/
(U/mL)		 总抗氧
化能力
T-AOC/
(mmol/L)		 丙二醛
MDA/
(nmol/mL)
色氨酸水平Try levels/%
0.31 85.06
±1.16a		 89.91
±1.28		 14.14
±0.51a		 6.28
±0.09a		 21.67
±0.28a		 858.33
±12.81a		 0.43
±0.02a		 2.34
±0.03c
0.62 87.46
±1.16a		 89.02
±1.28		 14.35
±0.51a		 6.71
±0.09b		 32.59
±0.28b		 869.32
±12.81a		 0.67
±0.02c		 1.71
±0.02b
0.94 99.88
±1.16b		 89.78
±1.28		 16.31
±0.51b		 7.57
±0.09c		 48.32
±0.28c		 971.58
±12.81b		 0.55
±0.02b		 1.59
±0.02a
蛋氨酸水平Met levels/%
0.66 77.51
±1.16a		 90.85
±1.28		 15.11
±0.51a		 6.31
±0.09a		 22.10
±0.28a		 856.77
±12.81a		 0.51
±0.02a		 1.87
±0.02
0.99 88.09
±1.16b		 88.62
±1.28		 17.90
±0.51b		 7.34
±0.09c		 45.86
±0.28b		 971.83
±12.81b		 0.67
±0.02b		 1.90
±0.02
1.33 78.81
±1.16a		 89.23
±1.28		 14.89
±0.51a		 6.72
±0.09b		 29.75
±0.28a		 870.62
±12.81a		 0.50
±0.02a		 1.86
±0.02
双因素方差分析Two-way ANOVA
色氨酸水平Try level * ns * * * * * *
蛋氨酸水平Met level * ns * * * * * ns
交互Interaction * ns * * * * * *

3 讨论

3.1 饲料中添加Try和Met对中华绒螯蟹生长性能和体成分的影响

研究表明,适量补充Try和Met对水产动物的摄食和生长性能具有一定的促进作用[13-14]。Try可通过影响脑中神经递质5-HT的含量和调控胃肠调节肽-胃饥饿素(ghrelin)的分泌来影响动物的采食量,促进肌肉生长,从而提高水产动物的生长性能[15-16]。Met不仅能改善饲料适口性,提高蛋白质沉积率,而且还能降低饵料系数,提高蛋白质效率[7,14]。本研究发现,虽然饲料中添加Try和Met对中华绒螯蟹有显著的促生长效果,但是交互作用不显著,这与Han等[17]对鱼类饲料中Try和Met水平研究的结果相似。随着Met水平的升高,对中华绒螯蟹生长有明显负作用,说明高水平Met对Try有拮抗作用。研究发现,饲料中过量的色氨酸对中国对虾(Fenneropenaeus chinensis)摄食量产生抑制作用[18]。而本研究中Try最高添加水平组未受影响且效果相对较好,分析原因一是添加水平适宜,二是还未达到最佳效果,后期进行接续试验加以完善。本研究还发现,不同生长发育期的中华绒螯蟹对Try和Met的最适需求量也不同,初始体重50.27 g的中华绒螯蟹对Try和Met最适需求量分别为0.94%和0.99%,比张佳鑫[6]研究Try最适需求量的结果(0.75%)略大,比叶金云等[5]研究Met最适需求量的结果(1.12%)略小。这说明中华绒螯蟹对氨基酸的需要量与个体大小、年龄、饲养方式、试验环境及基础饲料配方(蛋白质源的不同)等因素相关[19-20]。
在本试验中,补充外源Try和Met可明显提高中华绒螯蟹肌肉中粗蛋白质含量,且存在交互作用。分析原因是Try通过激活信号通路,使肌肉中雷帕霉素靶蛋白(TOR)和核糖体蛋白S6激酶1(S6K1)的基因表达量上调,激活后的TOR能够调控因子,多位点进行磷酸化,加快体内蛋白质合成,从而促使粗蛋白质含量的增加[21-22]。而Met的添加也可以重新建立氨基酸平衡,使合成代谢高于分解代谢,从而提高蛋白质的合成速度,有利于体内蛋白质的沉积。相似结果在鱼类和甲壳类中均有发现[7-8]。

3.2 饲料中添加Try和Met对中华绒螯蟹肝胰腺中消化酶活性的影响

大量研究表明,水产动物体内消化酶活性与饲料营养水平表现出一定程度的适应性,添加外源氨基酸可改善肠道形态结构、提高机体消化酶活性。张佳鑫[6]研究发现,饲料中添加Try可显著提高中华绒螯蟹肝胰腺中蛋白酶和脂肪酶活性;刘翰吉[23]研究发现,饲料中添加Try对三倍体虹鳟(Oncorhynchus mykiss)肠道胰蛋白酶、脂肪酶和淀粉酶活性都产生了显著影响。陈义方[24]在不同规格的凡纳滨对虾饲料中分别添加不同水平的Met,结果发现显著提高了凡纳滨对虾(Litopenaeus vannamei)肝胰腺中蛋白酶和淀粉酶活性。陈晴等[4]研究发现,补充适宜的Met可促进中华绒螯蟹幼蟹肝胰腺和肠道的蛋白酶和脂肪酶分泌,且显著提高幼蟹的生长和抗氧化性能。本研究中同样发现,Try和Met对中华绒螯蟹肝胰腺中消化酶活性有积极影响。值得注意的是,0.62% Try水平对4种消化酶活性均有显著影响,0.94% Try水平仅增强了蛋白酶活性,其具体原因有待进一步研究。

3.3 饲料中添加Try和Met对中华绒螯蟹免疫功能和抗氧化能力的影响

AKP是动物代谢过程中重要的调控酶,可促进中华绒螯蟹钙质吸收、甲壳素分泌及形成,加速蜕壳过程,从而提高存活率[25],血淋巴中的ACP和AKP还能与免疫因子一起进行体液免疫[26]。本试验中,饲料Try和Met水平分别为0.94%和0.99%时,中华绒螯蟹血清AKP活性最强,与生长性能的变化趋势相一致。笔者猜测AKP活性的上升促进了钙质吸取,从而有利于其蜕壳和生长。PO是酚氧化酶原激活系统的产物,是衡量机体免疫功能的重要指标。研究发现,在饲料中添加氨基酸可促进PO活性[27-28],本研究同样发现了Try和Met作为外源氨基酸,可以促进血清PO活性,从而提高生物体免疫功能。
T-AOC是反映机体抗氧化能力的综合性指标,分解和清除机体代谢过程中所产生的活性氧。SOD和CAT都是生物体内一类重要的抗氧化酶,SOD最先被自由基激活,然后将其转化为氧气(O2)和过氧化氢(H2O2),CAT则将细胞代谢产生的毒性物质H2O2迅速分解为水(H2O)和O2[29]。MDA含量是反映机体抗氧化潜在能力的重要参数,可以反映机体脂质过氧化速率和强度,也能间接反映组织过氧化损伤程度。本研究中,试验组的中华绒螯蟹血清SOD和T-AOC活性升高,MDA含量降低,表明饲料适量添加Try和Met提高了中华绒螯蟹抗氧化能力。研究发现,Try和Met在机体抗氧化性能方面发挥着积极作用[15,30]。Try代谢后产生褪黑素,作为一种非敏类抗氧化剂可以清除体内的自由基从而达到抗氧化的作用,而Met则是一种具有提高机体抗氧化能力的营养素,代谢时可产生具有抗氧化能力的有机物,通过氧化还原途径发挥抗氧化功能[31]。相似结果在凡纳滨对虾[27]、斜带石斑鱼(Epinephelus coioides)[32]与花鲈(Lateolabrax japonicus)[33]等研究中都有所发现。

4 结论

饲料中适当补充Try和Met可提高中华绒螯蟹的生长性能,Try和Met的添加水平及其交互作用影响了肌肉中粗蛋白质含量、肝胰腺中消化酶活性、非特异性免疫力和抗氧化能力。本试验条件下,饲料中添加0.94% Try和0.99% Met对中华绒螯蟹的上述指标促进效果最佳。

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