小刺猴头发酵浸膏对草鱼免疫功能及肠道菌群的影响

发布时间：2018-04-04 15:31

本文选题：小刺猴头菌发酵浸膏　切入点：草鱼　出处：《吉林农业大学》2017年硕士论文

【摘要】：本文研究了小刺猴头发酵浸膏[submerged fermentation concentrate of Herici um caput-medusae(Bull.:Fr.)Pers.,HFC]对草鱼血液生化指标、抗嗜水气单胞菌(Aeromonas hydrophila)感染能力、肠道形态结构及消化酶活力的影响。同时采用PCR-DGGE技术研究HFC对草鱼肠道菌群组成的影响。本试验共选用600条草鱼幼鱼随机分为4组,3个重复/组,50条草鱼/重复。投喂基础饲料(添加0 mg/kg)和试验饲料(400mg/kg、800mg/kg、1200mg/kg)分别记作H0组、H400组、H800组、H1200组。对草鱼进行45d的饲养试验,分别在15d、30d、45d采集血液样本,检测血清中白蛋白(ALB)、总蛋白(TP)含量,酸性磷酸酶(ACP)、总超氧化歧化酶(T-SOD)、溶菌酶(LSZ)、碱性磷酸酶(AKP)的活力。并采用酶联免疫吸附剂法检测(Enzyme-Linked Immunosorbent Assays,ELISA)45d血清中免疫球蛋白IgM、细胞因子白介素2(IL-2)、白介素10(IL-10)和肿瘤坏死因子α(TNF-α)含量。在饲喂试验结束后腹腔注射A.hydrophila进行感染试验,计算感染后96h存活率。试验结果表明,饲喂30d后H1200组较H0组的ALB、TP含量显著升高(p0.05)。各浓度添加组均能提高草鱼血清中T-SOD、ACP、LSZ、AKP活力((p0.05);与对照组相比,H1200组草鱼血清中IL-2、IL-10的含量增加(p0.05);且饲喂不同浓度HFC均能提高草鱼抗嗜水气单胞菌的存活率。采集饲喂15d、30d、45d各组肠道内食糜样品,利用PCR-DGGE技术对肠道菌群进行检测,结果显示草鱼肠道菌群主要细菌具有一定的多样性,主要菌群为厚壁菌门(Firmicutes)、变形菌门(Proteobacteria)、放线菌门(Actinobacteria)和疣微菌门(Verrucomicrobia)。大部分为革兰氏阴性细菌,少数为革兰氏阳性细菌,且减少有害菌的数量。采集15d、30d、45d各组肠道及内容物,检测HFC对草鱼消化功能的影响。采用切片方法,测量肠道隐窝深度、绒毛长度并计算V/C比值,结果表明HFC能促进前、后肠绒毛生长,与H0组相比前肠V/C比值增加明显(p0.05),而对后肠V/C比值作用不显著(p0.05)。各组添加浓度HFC均能显著提高胰蛋白酶(Trypsin)活力(p0.05),对淀粉酶(Amylas)活力影响不显著(p0.05),添加浓度800mg/kg、1200mg/kg HFC对脂肪酶(Lipase)活力有一定的抑制作用。综上所述,添加800~1200mg/kg的HFC对草鱼的免疫功能、肠道菌群及消化功能均有改善作用。
[Abstract]:This paper studies the small constituents of [submerged fermentation concentrate of hair yeast extract Herici um caput-medusae (Bull.: Fr.) Pers., HFC] on blood biochemical indexes of grass carp, anti Aeromonas hydrophila (Aeromonas hydrophila) infection, intestinal morphology and digestive enzyme activity. At the same time using PCR-DGGE technology to study the effects of HFC on intestinal flora of grass carp group composition. This experiment selected 600 grass carp were randomly divided into 4 groups, 3 replicates per group, 50 grass carp / repeat. Feeding basal diet (containing 0 mg/kg) and test (400mg/kg, 800mg/kg, 1200mg/kg feed) were recorded as H0 group, H400 group, H800 group, H1200 group. Grass carp 45d feeding trial, respectively in 15d, 30d, 45d and blood samples were collected to detect serum albumin (ALB), total protein content (TP), acid phosphatase (ACP), total superoxide dismutase (T-SOD), lysozyme (LSZ), alkaline phosphatase (AKP) activity. Using enzyme-linked immunosorbent assay (Enzyme-Linked Immunosorbent, Assays, ELISA) of immunoglobulin IgM in serum of 45d, interleukin 2 (IL-2), interleukin 10 (IL-10) and tumor necrosis factor alpha (TNF- alpha) content. In the feeding experiment after intraperitoneal injection of A.hydrophila infection test, calculate infection after the survival rate of 96h. The experimental results show that the ALB H1200 group than in H0 group after feeding 30d, TP content increased significantly (P0.05). Each concentration group can improve the T-SOD, grass carp LSZ, ACP in serum, the activity of AKP ((P0.05); compared with the control group, IL-2 group, grass carp H1200 in serum increased the content of IL-10 (P0.05); and with different concentrations of HFC could improve the survival rate of grass carp against Aeromonas hydrophila. Acquisition feeding 15d 30d, 45d groups of intestinal digesta samples to detect intestinal bacteria by using PCR-DGGE technology, results showed that the intestinal flora is mainly bacteria with grass carp The diversity of flora as Firmicutes, Proteobacteria (Firmicutes) (Proteobacteria), actinobacteria (Actinobacteria) and verrucomicrobia (Verrucomicrobia). The majority of gram negative bacteria, a few Gram-positive bacteria, and reduce the number of harmful bacteria. Acquisition 15d, 30d. 45d group and intestinal contents, to study the effect of HFC on digestive function. Grass carp uses the slice method, measurement of intestinal crypt depth and villus length and the ratio of V/C was calculated, the results show that HFC can promote the growth of intestinal villi, compared with H0 group, the V/ ratio of C increased significantly before (P0.05), and the ratio of V/C on intestinal function significant (P0.05). The concentration of HFC was significantly increased trypsin activity (Trypsin) (P0.05), the amylase (Amylas) activity had no significant effect (P0.05), the concentration of 800mg/kg, 1200mg/ kg HFC (Lipase) on lipase activity inhibition. In summary, the addition of 800~1200mg/kg HFC on immune function of grass carp, improve intestinal microflora and digestive function was found.

【学位授予单位】：吉林农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S965.112

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