不同年龄段神农架金丝猴肠道菌群组成和差异的研究

发布时间：2018-03-29 16:59

本文选题：神农架金丝猴　切入点：年龄　出处：《华中农业大学》2015年硕士论文

【摘要】：金丝猴作为国家一级珍稀保护动物,数量稀少,有极高的科研价值,现已被列为濒危物种。神农架金丝猴是金丝猴的一种,属于川金丝猴。我国政府已采取多种措施保护神农架金丝猴,人工补食计划是其中之一。人工补食计划的实施改变了神农架金丝猴原有的生活环境和饮食结构,必然会导致神农架金丝猴肠道菌群发生变化,由于肠道菌群与肠道健康密切相关,肠道菌群的失调有可能导致肠道疾病和其它非肠道疾病。因此,研究不同年龄阶段神农架金丝猴的肠道菌群组成,对监测金丝猴的肠道疾病和今后开展金丝猴专用人工饲料对肠道菌群的影响具有重要理论和实际指导意义。本研究共采集神农架金丝猴粪便样品47份。其中17份随机采集的样品用于变性梯度凝胶电泳技术分析,探索神农架金丝猴肠道菌群组成和多样性信息;另外30份样品来自三个不同年龄段的30只神农架金丝猴,用于16S r DNA的V4区高通量测序分析,得到三个年龄段神农架金丝猴肠道菌群组成和差异的信息,并将测得结果与其他物种的肠道菌群进行了比较分析。取得了如下结果:1.17份随机采集样品的变性梯度凝胶电泳分析结果表明:不同个体之间肠道菌群的多样性存在差异;主要优势菌门及比例:厚壁菌门(Firmicutes,55.6%)和拟杆菌门(Bacteroidetes,13.9%)。2.30份不同年龄段粪样高通量测序分析结果表明:神农架金丝猴肠道主要优势菌门为厚壁菌门(Firmicutes,20.96%-30.29%)和拟杆菌门(Bacteroidetes,29.85%-39.46%);0-3岁组(Y1),5-8岁组(Y2),12岁以上组(Y3)肠道菌群含OTUs的数目依次为1469,1381,1931;菌门数目依次为16,16,19;属的数目依次为166,157,190;从不同年龄段肠道菌群OUTs数目比较可以看出:12岁以上组肠道菌群所含种类最多,肠道菌群最为丰富;UPGMA聚类分析表明0-3岁组(Y1)和5-8岁组(Y2)肠道菌群组成较为接近。3.神农架金丝猴肠道菌群与其他物种的比较分析结果显示,厚壁菌门和拟杆菌门是神农架金丝猴肠道和人、非人灵长类肠道共同的优势菌门,但在两者在3个物种肠道内所含的比例有差异(其中金丝猴为:57.88%-62.17%;人为:90%以上;非人灵长类70.50%-98.30);变形菌门在神农架金丝猴和人肠道中所占比例有较大的差异;螺旋体门是神农架金丝猴肠道的优势菌门,在人、非人灵长类肠道中为非优势菌门;神农架金丝猴肠道厚壁菌门以梭菌纲为主,且含有反刍动物特有的纤维杆菌门;神农架金丝猴的肠道中存在反刍动物特有的甲烷短杆菌。
[Abstract]:As one of the country's first class protected animals, the golden monkey is rare in number and of great scientific research value, and has been listed as an endangered species. The Shennongjia golden monkey is a kind of golden monkey. Belong to Sichuan golden monkey. Our government has taken many measures to protect Shennongjia golden monkey, among which artificial supplementary feeding plan is one of them. The implementation of artificial supplementary feeding program has changed the original living environment and diet structure of Shennongjia golden monkey. This will inevitably lead to changes in the intestinal flora of the Shennongjia golden monkey. Because the intestinal flora is closely related to intestinal health, the imbalance of the intestinal flora may lead to intestinal diseases and other non-intestinal diseases. The composition of intestinal flora of Shennongjia golden monkey at different ages was studied. It is of great theoretical and practical significance to monitor the intestinal diseases of golden monkey and to develop the effect of artificial diet for golden monkey on intestinal flora. In this study, 47 fecal samples of Shennongjia golden monkey were collected. 17 of them were collected from the faeces of Shennongjia golden monkey. Random samples were used for denaturing gradient gel electrophoresis (DGGE) analysis. To explore the composition and diversity information of intestinal flora of Shennongjia golden monkey, another 30 samples were collected from 30 Shennongjia golden monkeys of different ages and used for high-throughput sequencing analysis of V4 region of 16s r DNA. The composition and difference of intestinal flora of three age groups of Shennongjia golden monkey were obtained. The results were compared with those of other species. The following results were obtained: 1.17 randomly collected samples were analyzed by denaturing gradient gel electrophoresis. The results showed that the diversity of intestinal flora was different among different individuals. Main dominant phylum and proportion: Firmicutesa (55.65.65.65.65.65.65.65.65) and Bacteroidetes 13.99.2.30 fecal samples of different age groups high throughput sequencing analysis showed that the main dominant bacteria in the intestinal tract of the golden monkey in Shennongjia were Firmicutes20.96-30.29292929) and Bacteroidetes 29.85-39.460-3 years old. The number of OTUs in intestinal microflora was 1469, 1381 and 1931, respectively, and the number of bacteria was 166157190 and 166157190 in turn. The number of OUTs in intestinal flora of different age groups was the most, and the number of OUTs in intestinal flora over 12 years of age was the most, and the number of genera was 166157190.These results showed that the number of intestinal microflora was the most in group over 12 years of age, and the number of bacteria in intestinal flora of different age groups was more than 12 years old. The results of UPGMA cluster analysis showed that the composition of intestinal flora of 0-3 years old group and 5-8 year old group was close to that of 3.The results of comparison between Shennongjia golden monkey and other species showed that the intestinal microflora of Shennongjia golden monkey was similar to that of other species. The phylum and Bacteroides phylum are the common dominant phylum of intestinal tract of Shennongjia golden monkey and human and non-human primates, but the proportion of them in the intestinal tract of three species is different (the golden monkey is: 57.88-62.17g), the proportion of human being above 90%; The proportion of Proteus phylum in the intestine of golden monkey of Shennongjia is different from that of human, and that of spirochetes is the dominant bacteria in the intestinal tract of Shennongjia golden monkey, which is non-dominant in human and non-human primate intestines. Shenongjia golden monkey intestinal clostridium is mainly Clostridium, and contains ruminant specific cellulose bacilli, Shennongjia golden monkey intestinal tract of ruminant unique methanobacterium.
【学位授予单位】：华中农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S852.6

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