植物蛋白质饲料中添加巨大芽孢杆菌对杂交鲟幼鱼生长、免疫的影响及机理探讨

发布时间：2018-05-05 01:29

本文选题：巨大芽孢杆菌 + 杂交鲟　；参考：《上海海洋大学》2017年硕士论文

【摘要】：本研究旨在探讨植物蛋白质饲料基础上添加不同水平的巨大芽孢杆菌对杂交鲟(Acipenser schrenckii♀×Acipenser baeri♂)幼鱼生长、免疫的影响。以鱼粉(FM)组和大豆浓缩蛋白(PM)组为对照组,试验Ⅰ、Ⅱ、Ⅲ、Ⅳ组分别在PM组基础上添加106 cfu/g市售枯草芽孢杆菌和105、106、107 cfu/g本实验室筛选鉴定的巨大芽孢杆菌。驯养结束后选取750尾平均体重为(12.87±0.65)g的杂交鲟幼鱼,随机分成6组,每组设置5个重复,每个重复25尾鱼。试验期间保持自然光照,水温控制在21±0.5℃,溶氧大于5mg/L。每天人工分别在7:00、12:00、17:00饱食投喂3次,并记录投喂量,定期检测水质,换水、消毒,确保水质良好。试验养殖周期为8周。试验结果如下:(1)FM组终末重和增重率显著高于其他各组(P0.05),且试验Ⅱ组显著高于PM组(P0.05);FM组和试验Ⅱ组饲料系数显著低于其他各组,且试验Ⅰ组显著低于PM组(P0.05);FM组和试验Ⅱ组蛋白质效率显著高于其他各组,且试验Ⅰ组显著高于PM组(P0.05);饲料中巨大芽孢杆菌对鱼体肥满度、干物质、粗脂肪、粗蛋白和粗灰分无显著影响(P0.05)。(2)FM组和试验Ⅰ、Ⅱ组血清和肝脏的溶菌酶(LSZ)含量、碱性磷酸酶(AKP)活性显著高于PM组(P0.05);试验Ⅰ、Ⅲ组血清髓过氧化物酶(MPO)活性显著高于PM组(P0.05);试验Ⅱ组肝脏MPO活性显著高于PM组和试验Ⅳ组(P0.05);FM组和各试验组嗜水气单胞菌攻毒试验的死亡率均低于PM组。(3)FM组及试验Ⅱ、Ⅲ、Ⅳ组血清球蛋白(GLB)、总蛋白(TP)含量显著高于PM组(P0.05);FM组和试验Ⅱ组血清尿酸(UA)含量显著低于PM组(P0.05);FM组和试验Ⅲ组血清谷丙转氨酶(ALT)活性显著低于PM组(P0.05);试验Ⅰ、Ⅱ、Ⅲ组血清谷草转氨酶(AST)活性显著低于PM组(P0.05);试验Ⅲ组血糖(Glu)含量显著低于PM组(P0.05)。(4)试验Ⅱ组血清、肝脏和十二指肠总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)、抗超氧阴离子自由基(ASAFR)活性和谷胱甘肽(GSH)含量显著高于PM组(P0.05),试验Ⅰ组血清过氧化氢酶(CAT)活性显著高于PM组(P0.05),试验Ⅲ、Ⅳ组血清和肝脏黄嘌呤氧化酶(XOD)活性显著低于PM组(P0.05),试验Ⅰ、Ⅱ、Ⅲ组幽门盲囊GSH含量、T-AOC活性和SOD活性显著高于PM组(P0.05),且与FM组相比差异均不显著(P0.05)。(5)试验Ⅰ、Ⅱ组胃、肠道的蛋白酶活性显著高于PM组(P0.05);FM组和试验Ⅲ组胃、幽门盲囊、肠道的淀粉酶活性显著高于PM组(P0.05);试验Ⅱ组胃淀粉酶活性显著高于其它各组(P0.05);FM组和试验Ⅱ组胃、幽门盲囊、肠道的脂肪酶活性显著高于PM组(P0.05)。(6)试验Ⅰ、Ⅱ组脾脏NF-kB、HSP90和HSP70基因的相对表达量显著高于PM组和FM组(P0.05);试验Ⅱ组肠道NF-k B、HSP70、MyD88和HSP90基因相对表达量显著高于PM组(P0.05)。(7)FM组和试验Ⅱ、Ⅲ、Ⅳ组十二指肠绒毛高度、肌层厚度显著高于PM组(P0.05);试验Ⅱ、Ⅲ组十二指肠褶皱深度显著高于PM组(P0.05);FM组及试验Ⅰ、Ⅱ组瓣肠绒毛高度、褶皱深度和肌层厚度显著高于PM组(P0.05)。(8)试验Ⅰ组和Ⅲ组Ace值高于其他各组;试验Ⅰ组Chao值和Shannon值高于其他各组;试验Ⅱ、Ⅲ、Ⅳ组Simpson值均高于PM组。在门类学水平上,梭杆菌门(Fusobacteria)和厚壁菌门(Firmicutes)是杂交鲟幼鱼肠道内含物菌落中最主要的菌门;随着巨大芽孢杆菌添加量的增加,厚壁菌门的相对丰度逐渐升高,梭杆菌门和变形菌门(Proteobacteria)的相对丰度逐渐降低。综合以上结果:在植物蛋白质饲料中添加巨大芽孢杆菌可以提高杂交鲟幼鱼增重率和蛋白质效率,提高消化酶活性,改善肠道形态,调节肠道菌群结构;还可以提高鱼体抗氧化能力,非特异性免疫能力及免疫相关基因相对表达量;添加105 cfu/g巨大芽孢杆菌对杂交鲟幼鱼影响效果较好。
[Abstract]:The aim of this study was to explore the effects of different levels of Bacillus megigantoin on the growth and immunity of the hybrid sturgeon (Acipenser schrenckii Acipenser baeri baeri), with fish meal (FM) group and soybean protein concentrate (PM) group as the control group. The experiment I, II, III and IV group added 106 cfu/g city on the basis of PM group, respectively. Bacillus subtilis and 105106107 cfu/g laboratory screened Bacillus giant bacillus. After the end of domestication, the hybrid sturgeon young with 750 tails (12.87 + 0.65) g was randomly divided into 6 groups, each set 5 repetitions, each repeated 25 fish. During the test, the natural light was kept, the water temperature was controlled at 21 + 0.5 C, and the dissolved oxygen was greater than 5mg/L. Every day, 3 times were fed on 7:00,12:00,17:00, and the feeding amount was recorded, the water quality was detected regularly, water changed, and the water quality was good. The experimental breeding period was 8 weeks. The results of the test were as follows: (1) the end weight and weight gain rate of group FM were significantly higher than that of other groups (P0.05), and the test group II was significantly higher than that of group PM (P0.05); group FM and experimental group II feed The coefficient of the experimental group I was significantly lower than that of the PM group (P0.05), and the protein efficiency of the FM group and the test group was significantly higher than that of the other groups, and the test group I was significantly higher than that of the PM group (P0.05). There was no significant effect of Bacillus giant Bacillus on fish body fullness, dry matter, fat, crude protein and crude ash (P0.05). (2) FM group and test In group I, the content of serum and liver lysozyme (LSZ) and alkaline phosphatase (AKP) activity were significantly higher than that of group PM (P0.05), and the activity of serum myeloperoxidase (MPO) in group III was significantly higher than that in group PM (P0.05), and the activity of MPO in group II was significantly higher than that in group PM and group IV (P0.05), and the death of Aeromonas hydrophila in FM group and test group was dead. The rate of death was lower than that of the PM group. (3) the content of serum globulin (GLB) and total protein (TP) in group FM and group IV was significantly higher than that in the PM group (P0.05); the serum uric acid (UA) content in the FM group and test group II was significantly lower than that in the PM group (P0.05), and the activity of seropropane (ALT) in the FM and test group was significantly lower than that in the group (P0.05). The activity of enzyme (AST) was significantly lower than that in group PM (P0.05); the content of blood glucose (Glu) in group III was significantly lower than that in group PM (P0.05). (4) the total antioxidant capacity (T-AOC), superoxide dismutase (SOD), anti superoxide anion free radical (ASAFR) activity and glutathione (GSH) content of the liver and duodenum, and the content of glutathione (GSH) were significantly higher than that of PM group (P0.05). The activity of catalase (CAT) was significantly higher than that in group PM (P0.05). The activity of serum and liver xanthine oxidase (XOD) in group IV and liver was significantly lower than that in PM group (P0.05). The GSH content of the pyloric caeca in group I, II, and group III was significantly higher than that of the PM group (P0.05). (5) test I, group II stomach and intestinal tract (5) The activity of protease was significantly higher than that of the PM group (P0.05); the activity of amylase in gastric, pyloric caecum and intestinal tract was significantly higher in group FM and test group III than in group PM (P0.05); the activity of gastric amylase in group II was significantly higher than that in other groups (P0.05); the activity of lipase in gastric, pyloric caecum and intestinal tract in group FM and test II group was significantly higher than that in group PM (P0.05). (6) test I, group II spleen The relative expression of NF-kB, HSP90 and HSP70 genes was significantly higher than that in group PM and FM (P0.05). The relative expression of NF-k B, HSP70, MyD88 and HSP90 genes in the intestinal tract was significantly higher than that in the PM group. (7) the height of the duodenal villus and the thickness of the muscular layer were significantly higher than that in the group and test group and test II, III and IV group. Significantly higher than group PM (P0.05), group FM and test I, group II, the height of intestinal villus, fold depth and muscularis thickness were significantly higher than group PM (P0.05). (8) the test I group and group III Ace value is higher than the other groups; test I group Chao value and Shannon value are higher than the other groups; test II, III, IV group Simpson values are higher than the PM group. At the level of the Clostridium, Clostridium Fusobacteria and Firmicutes are the most important bacteria in the intestinal bacterial colonies of the hybrid sturgeon. With the increase of Bacillus megaciformis, the relative abundance of the thick wall fungus increases gradually, and the phase of the Clostridium and deformable bacteria gate (Proteobacteria) decreases gradually. The addition of Bacillus giant Bacillus could increase the weight gain and protein efficiency of the young sturgeon, improve the digestive enzyme activity, improve the intestinal morphology, regulate the intestinal flora structure, and improve the antioxidant capacity of the fish body, the non specific immunity and the relative expression of the immune related genes, and add 105 cfu/g Bacillus megigantobacilli to the hybrid sturgeon. The effect of fish is better.

【学位授予单位】：上海海洋大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S965.215

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