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摘要：肠道微生物群在鸭主要生产性能、健康和免疫力方面担任重要角色。了解肠道微生物群落组成的拓扑差异对提供关于生物群体的每个成员的功能知识是至关重要的，这些微生物群对鸭的主要机能的生理学文护很重要。本研究采用高通量测序技术深入分析了鸭盲肠的微生物群区系。所用的盲肠是来自正常饲养的三周龄北京鸭的五个个体。提取样品总DNA 后，根据设计得到的16SrRNA 高变区的 V3+V4 区域合成引物，合并引物接头，进行 PCR 扩增并对其产物进行纯化、定量和均一化形成测序文库，建好的文库先进行文库质检，质检合格的文库用 Illumina MiSeq PE300 进行测序。通过实验我们观察到经过测序数据质控、序列优化、OUT聚类、物种分类与丰度分析、单样品多样性分析、多样品多样性分析共得出1447个OUT，并得出样品在各分类水平上的物种组成及不同物种所占的比例：门分类水平上拟杆菌门（55%-74%）是最丰富的物种；在属分类水平上拟杆菌属（43%-61%）是最丰富的物种。并且进一步了解了鸭的不同肠道微生物有着明显不同的功能和角色。最后我们得出以下结论：从门到属，随着分类标准越精确，微生物菌群的类别也就越多。但拟杆菌仍然占较大的比值。基于肠道微生物群落的组成，使用有利的细菌比如说有益于主要机能的益生菌对未来调制肠道是至关重要的。26550
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毕业论文关键词:北京鸭；肠道微生物群；盲肠。
The analysis of Peking duck cecum gut microflora
Abstract: Duck gut microbiota has important roles in host performance, health and immunity. Understanding the topological difference in gut microbial community composition is crucial to provide knowledge on the functions of each members of microbiota to the physiological maintenance of the host. This study used High-throughput sequencing technologies to analyze the microbiota fauna of the duck cecum. The cecum used is from five three weeks Peking ducks which is fed normally. After the extraction of total DNA samples, we synthetized primers according to the V3 + V4 hyper-variable region of 16S rRNA .Then we conducted PCR amplification and the purification of the PCR product. Next, we formed the sequencing library. After sequencing, sequence optimization, OUT clustering, analysis of classification and abundance of species persity, single sample analysis, analysis of the persity of many samples, we can get 1447 OUT totally, and find that the species composition of samples and proportion in all samples is different: From the classification level in Phylum, we can find that bacteroidetes is one of the most abundant species (55% -74%); From the classification level in Genus, we can find that bacteroides is one of the most abundant species (43% -61%). And we learn that different duck gut microbes have obvious difference between function and role. Finally, we can conclude that from the phylum to genus, as the more precise classification standard, we can detected the more microbial flora of the category. The ratio of bacteroides still represents a significant proportion. Based on the composition of the microbial community, future gut modulation with beneficial bacteria such as probiotics may benefit the host.
Key words: Peking duck; Gut microbiota; Caecum.
目录
引言 1
1 材料与方法 1
1.1实验流程图 1
1.2信息分析流程 2
1.3鸭和采样 3
1.4 DNA的提取 4
1.5 16S rRNA 高变区的 V3+V4 区域基因扩增和Illumina测序 4
1.6 生物信息学分析 4
2 结果 5
2.1 测序数据质控 5
2.2 序列优化 5
2.2.1 Reads 拼接成 Tags 5
2.2.2 Tags 优化及统计 6