基于高通量测序的健康与子宫内膜炎奶牛阴道菌群差异性研究
发布时间：2018-10-08 21:42
【摘要】：奶牛阴道菌群在维持其生殖道正常生理功能和预防疾病过程中起着非常重要的作用,阴道菌群的研究对诊治子宫内膜炎至关重要。目前,虽已有相关报道,但我们仍对阴道菌群的多样性和动态变化知之甚少。本试验为了深入研究奶牛阴道菌群的复杂变化和其与子宫内膜炎的关系,运用高通量测序对健康和子宫内膜炎奶牛产后不同时间段的阴道菌群进行比较研究。选取长春市地区的荷斯坦奶牛为实验动物,采集3头健康奶牛和3头患子宫内膜炎奶牛产后6-8d和34-36d的阴道黏液。通过提取样品中细菌总DNA、扩增细菌16S rDNA v3区片段、建立高通量测序文库,然后对测序文库进行高通量测序,最后通过生物学分析比较健康与患子宫内膜炎奶牛产后不同时间段阴道菌群的结构差异。本试验通过对样品高通量(Illumina MiSeq)测序,经过数据处理,最终得到710382条有效序列,再经生物信息学分析发现20个已知菌门的165已知菌属,还有一些未知菌类,每组样品在物种水平分类上的菌群数量、种类及丰度均存在差异。厚壁菌门(Firmicutes)为健康奶牛和患子宫内膜炎奶牛产后不同时间段阴道内共有的优势菌门,放线菌门(Actinobacteria)为产后34-36d的上述两种奶牛阴道内的优势菌门,梭杆菌门(Fusobacteria)为产后6-8d患病奶牛阴道的优势菌门。在属的分类水平上我们发现健康奶牛和患病奶牛在产后34-36d阴道内共有优势菌属为梭菌属(Clostridium),创伤球菌属(Helcococcus)和梭杆菌属(Fusobacterium)为产后6-8d患病奶牛阴道内优势菌属,健康奶牛产后6-8d并没有发现优势菌属。本试验经研究表明奶牛阴道菌群的构建是一个动态演变的过程,产后不同阶段健康奶牛和患子宫内膜炎奶牛阴道菌群的复杂度随产后时间的延长不断增加,并发现是在产后34-36d健康奶牛阴道内的菌群数量和种类均为最多。总体上看,本试验证明了奶牛阴道内有其独特的菌群结构,发现健康奶牛和患子宫内膜炎奶牛的阴道内细菌群落构的差异性,并随时间呈动态变化。
[Abstract]:The vaginal flora plays an important role in maintaining the normal physiological function of the reproductive tract and preventing the disease. The study of the vaginal flora is very important to the diagnosis and treatment of endometritis. Although there have been reports, we still know little about the diversity and dynamic changes of vaginal flora. In order to study the complex changes of vaginal flora and its relationship with endometritis in dairy cows, a comparative study was carried out on the vaginal flora of healthy and endometritis cows at different periods after delivery by high-throughput sequencing. Holstein cows in Changchun were selected as experimental animals. Vaginal mucus was collected from 3 healthy cows and 3 cows with endometritis at 6-8 and 34-36 days postpartum. The 16s rDNA v3 fragment was amplified by total DNA, from the sample, and the high-throughput sequencing library was established, and then the high-throughput sequencing was carried out. Finally, biological analysis was carried out to compare the structure of vaginal flora between healthy cows and cows with endometritis. Through high throughput (Illumina MiSeq) sequencing of the sample, 710382 effective sequences were obtained through data processing, and then the bioinformatics analysis revealed that there were 20 known genera of 165known bacteria, and some unknown fungi. There were differences in the number, species and abundance of microflora in the taxonomy at species level in each group. (Firmicutes) was the dominant bacteria in vagina of healthy cows and cows with endometritis at different times after delivery, and (Actinobacteria) was the dominant bacteria in the vagina of the above two kinds of cows after 34-36 days postpartum. Clostridium clostridium (Fusobacteria) is the dominant bacterium in the vagina of dairy cows after 6-8 days postpartum. At the taxonomic level, we found that the dominant bacteria in vagina of healthy cows and sick cows were Clostridium (Clostridium), (Helcococcus) and (Fusobacterium), which were dominant in vagina of healthy cows and sick cows after 6-8 days postpartum. No dominant genus was found on 6-8 days postpartum in healthy cows. The study shows that the construction of vaginal flora is a dynamic evolution process, and the complexity of vaginal flora of healthy cows and cows suffering from endometritis increases with the extension of postpartum time. It was found that the number and species of bacteria in vagina of healthy dairy cows were the highest in 34-36 days postpartum. On the whole, this experiment proved that there is a unique flora structure in the vagina of dairy cows, and found the difference of the bacterial community structure between healthy cows and cows suffering from endometritis, and showed dynamic changes with time.
【学位授予单位】：吉林农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S858.23
本文编号：2258365
[Abstract]:The vaginal flora plays an important role in maintaining the normal physiological function of the reproductive tract and preventing the disease. The study of the vaginal flora is very important to the diagnosis and treatment of endometritis. Although there have been reports, we still know little about the diversity and dynamic changes of vaginal flora. In order to study the complex changes of vaginal flora and its relationship with endometritis in dairy cows, a comparative study was carried out on the vaginal flora of healthy and endometritis cows at different periods after delivery by high-throughput sequencing. Holstein cows in Changchun were selected as experimental animals. Vaginal mucus was collected from 3 healthy cows and 3 cows with endometritis at 6-8 and 34-36 days postpartum. The 16s rDNA v3 fragment was amplified by total DNA, from the sample, and the high-throughput sequencing library was established, and then the high-throughput sequencing was carried out. Finally, biological analysis was carried out to compare the structure of vaginal flora between healthy cows and cows with endometritis. Through high throughput (Illumina MiSeq) sequencing of the sample, 710382 effective sequences were obtained through data processing, and then the bioinformatics analysis revealed that there were 20 known genera of 165known bacteria, and some unknown fungi. There were differences in the number, species and abundance of microflora in the taxonomy at species level in each group. (Firmicutes) was the dominant bacteria in vagina of healthy cows and cows with endometritis at different times after delivery, and (Actinobacteria) was the dominant bacteria in the vagina of the above two kinds of cows after 34-36 days postpartum. Clostridium clostridium (Fusobacteria) is the dominant bacterium in the vagina of dairy cows after 6-8 days postpartum. At the taxonomic level, we found that the dominant bacteria in vagina of healthy cows and sick cows were Clostridium (Clostridium), (Helcococcus) and (Fusobacterium), which were dominant in vagina of healthy cows and sick cows after 6-8 days postpartum. No dominant genus was found on 6-8 days postpartum in healthy cows. The study shows that the construction of vaginal flora is a dynamic evolution process, and the complexity of vaginal flora of healthy cows and cows suffering from endometritis increases with the extension of postpartum time. It was found that the number and species of bacteria in vagina of healthy dairy cows were the highest in 34-36 days postpartum. On the whole, this experiment proved that there is a unique flora structure in the vagina of dairy cows, and found the difference of the bacterial community structure between healthy cows and cows suffering from endometritis, and showed dynamic changes with time.
【学位授予单位】：吉林农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S858.23
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