基于高通量測序的健康與子宮內(nèi)膜炎奶牛陰道菌群差異性研究
發(fā)布時間：2018-10-08 21:42
【摘要】：奶牛陰道菌群在維持其生殖道正常生理功能和預(yù)防疾病過程中起著非常重要的作用,陰道菌群的研究對診治子宮內(nèi)膜炎至關(guān)重要。目前,雖已有相關(guān)報道,但我們?nèi)詫﹃幍谰旱亩鄻有院蛣討B(tài)變化知之甚少。本試驗(yàn)為了深入研究奶牛陰道菌群的復(fù)雜變化和其與子宮內(nèi)膜炎的關(guān)系,運(yùn)用高通量測序?qū)】岛妥訉m內(nèi)膜炎奶牛產(chǎn)后不同時間段的陰道菌群進(jìn)行比較研究。選取長春市地區(qū)的荷斯坦奶牛為實(shí)驗(yàn)動物,采集3頭健康奶牛和3頭患子宮內(nèi)膜炎奶牛產(chǎn)后6-8d和34-36d的陰道黏液。通過提取樣品中細(xì)菌總DNA、擴(kuò)增細(xì)菌16S rDNA v3區(qū)片段、建立高通量測序文庫,然后對測序文庫進(jìn)行高通量測序,最后通過生物學(xué)分析比較健康與患子宮內(nèi)膜炎奶牛產(chǎn)后不同時間段陰道菌群的結(jié)構(gòu)差異。本試驗(yàn)通過對樣品高通量(Illumina MiSeq)測序,經(jīng)過數(shù)據(jù)處理,最終得到710382條有效序列,再經(jīng)生物信息學(xué)分析發(fā)現(xiàn)20個已知菌門的165已知菌屬,還有一些未知菌類,每組樣品在物種水平分類上的菌群數(shù)量、種類及豐度均存在差異。厚壁菌門(Firmicutes)為健康奶牛和患子宮內(nèi)膜炎奶牛產(chǎn)后不同時間段陰道內(nèi)共有的優(yōu)勢菌門,放線菌門(Actinobacteria)為產(chǎn)后34-36d的上述兩種奶牛陰道內(nèi)的優(yōu)勢菌門,梭桿菌門(Fusobacteria)為產(chǎn)后6-8d患病奶牛陰道的優(yōu)勢菌門。在屬的分類水平上我們發(fā)現(xiàn)健康奶牛和患病奶牛在產(chǎn)后34-36d陰道內(nèi)共有優(yōu)勢菌屬為梭菌屬(Clostridium),創(chuàng)傷球菌屬(Helcococcus)和梭桿菌屬(Fusobacterium)為產(chǎn)后6-8d患病奶牛陰道內(nèi)優(yōu)勢菌屬,健康奶牛產(chǎn)后6-8d并沒有發(fā)現(xiàn)優(yōu)勢菌屬。本試驗(yàn)經(jīng)研究表明奶牛陰道菌群的構(gòu)建是一個動態(tài)演變的過程,產(chǎn)后不同階段健康奶牛和患子宮內(nèi)膜炎奶牛陰道菌群的復(fù)雜度隨產(chǎn)后時間的延長不斷增加,并發(fā)現(xiàn)是在產(chǎn)后34-36d健康奶牛陰道內(nèi)的菌群數(shù)量和種類均為最多。總體上看,本試驗(yàn)證明了奶牛陰道內(nèi)有其獨(dú)特的菌群結(jié)構(gòu),發(fā)現(xiàn)健康奶牛和患子宮內(nèi)膜炎奶牛的陰道內(nèi)細(xì)菌群落構(gòu)的差異性,并隨時間呈動態(tài)變化。
[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.
【學(xué)位授予單位】：吉林農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S858.23
本文編號：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.
【學(xué)位授予單位】：吉林農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S858.23
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