蒙古马肠道细菌多样性的研究

发布时间：2018-05-12 21:21

本文选题：蒙古马 + 肠道细菌　；参考：《内蒙古农业大学》2015年硕士论文

【摘要】：蒙古马是我国著名的地方品种,有着优秀的品种遗传资源。已被录入我国保种名录中。比其它马种蒙古马具有耐粗饲特性,抓膘迅速,掉膘缓慢。这也是蒙古马长期在恶劣环境条件下进化出来的生存能力。蒙古马的耐粗饲特性除了遗传因素,必然与其特殊的肠道微生物区系相关。为了了解其中微生物作用关系,本实验对蒙古马和纯血马肠道细菌多样性初步研究。也为功能细菌的挖掘和利用提供基础。本实验以纯血马和不同饲养条件下的蒙古马新鲜粪便为实验样品,用生物试剂盒直接从粪样中提取细菌基因组DNA,使用PCR技术从细菌总DNA中扩增出16SrDNA V3区序列并采用变性梯度凝胶电泳分离不同细菌的基因序列。用Quantity One软件对胶图进行分析,同时切胶回收优势条带进行克隆测序。将得到的测序在基因库数据里进行Blast比对判断菌种属性。实验主要结果以下：第一、蒙古马和纯血马肠道细菌群落结构比较中,DGGE胶图显示不同位置的条带有53个。选择41个优势条带回收测序鉴定中与NCBI数据库细菌16S rDNAV3区序列相似性96%以上的仅有16个克隆。在蒙古马两种不同饲养条件下的肠道细菌结构变化的研究中,DGGE胶图中显示共有50个条带,选40个优势条带回收测序,结果序列相似性96%以上有17个克隆。因此同源性较低。两次实验中分别未报道的优势细菌占总细菌的约61%和58%。第二、利用Quantity One软件分析出的DGGE图谱识别的条带数目和群落多样性指标来看,纯血马肠道优势菌比蒙古马肠道优势菌更丰富多样。聚类分析结果蒙古马和纯血马各自聚成一支,品种之间肠道优势菌相似性仅为0.33。品种内相似性都大于品种间相似性。蒙古马7月份草原上放牧时的肠道优势菌比3月份圈养时的肠道优势菌更丰富多样。聚类分析后蒙古马3月份和7月份各自聚成一支,之间相似性为0.54。蒙古马吃干草情况下的种内相似性大于吃青草时的。第三、蒙古马和纯血马肠道内共有的优势菌有栖瘤胃普雷沃氏菌、溶纤维丁酸弧菌、微黄棒状杆菌、黄化瘤胃球菌和枯草芽孢杆菌,以上菌种中除了枯草芽孢杆菌都具有降解纤维功能。枯草芽孢杆菌是多功能益生菌。蒙古马肠道内特有的优势菌有瘤胃球菌属、解纤维梭菌、化糖梭状芽胞杆菌。其中前两者是较强的纤维降解菌。纯血马肠道内特有的优势菌有巴氏梭状芽胞杆菌、嗜胺梭菌、氧化还原真杆菌、活泼瘤胃球菌、共生梭菌、乳酸双歧杆菌、牛链球菌、炭疽芽孢杆菌。蒙古马7月份草原上放牧比3月份圈养条件,肠道内新增优势菌有解纤维醋弧菌、直肠真杆菌、梭状梭菌、一致粪球菌、短真杆菌。相反微黄棒状杆菌和枯草芽孢杆菌在蒙古马放牧吃青草情况下变成不是优势条带。蒙古马两种不同饲养条件下肠道主要纤维菌的种类变化不大。
[Abstract]:Mongolia horse is a famous local breed in China and has excellent genetic resources. It has been recorded in the list of Chinese species. Compared with other horses, Mongolia horses have the characteristics of rough feeding, fast fat and slow fat. This is also the survival ability of Mongolia horses in the long period of bad environment. The resistance to coarse feeding of Mongolia horses is in addition to genetic causes In order to understand the relationship between the microorganism and the microbial flora of the Mongolia horse and the pure blood horse, the preliminary study on the diversity of the intestinal bacteria in the Mongolia horse and the pure blood horse also provided the basis for the excavation and utilization of the functional bacteria. The experiment was used in this experiment. The experimental samples of the fresh manure of the horse and the Mongolia horse under different feeding conditions were used as the experimental samples. The biological kit extracted bacterial genome DNA directly from the fecal sample, amplified the sequence of 16SrDNA V3 region from the total DNA of bacteria and separated the gene sequences of different bacteria by denaturing gradient gel electrophoresis with the PCR technique. The gel was analyzed with Quantity One software, and the gel was sequenced and sequenced. The Blast comparison of the gene pool data was used to determine the bacterial properties. The main results were as follows: first, in the comparison of the bacterial community structure of the Mongolia horse and the pure blood horse intestinal bacteria, the DGGE map showed that there were 53 different positions in different locations. The only 41 dominant strip recovery sequencing identification was more than 96% of the similarity of the NCBI database bacterial 16S rDNAV3 region sequence. There were 16 clones. In the study of the intestinal bacterial structure changes in two different feeding conditions of Mongolia horse, there were 50 bands in the DGGE map, and 40 dominant bands were recovered and sequenced. The result sequence similarity was more than 96%. Therefore, the homology was low. The dominant bacteria in the two experiments were about 61% of the total bacteria. And 58%. second, with the number of bands and community diversity identified by Quantity One software, the dominant bacteria in the intestinal tract were more diverse than the Mongolia horse enteric predominant bacteria. The results of cluster analysis showed that the Mongolia horse and the pure blood horse were together in one branch, and the similarity of the intestinal dominant bacteria among the varieties was only within the internal similarity of the 0.33. varieties. In July, the intestinal dominant bacteria in the pasture of Mongolia horse were more diverse than those of the enteric predominant bacteria in the March captivity. After cluster analysis, the Mongolia horses were gathered in March and July, and the similarity between the 0.54. Mongolia horse and the hay was greater than that of the green grass. Third, Mongolia. The dominant bacteria in the gut of the horse and the pure blood horse have the predominant bacteria in the rumen, the strains of Vibrio butyrate, the Corynebacterium yellowish, the Yellow rumen coccus and Bacillus subtilis, and the Bacillus subtilis all have the function of degrading the fibers. Bacillus subtilis is a probiotic probiotic. The dominant bacteria in the intestinal tract of Mongolia horse are Ruminal coccus, Clostridium difficile and Clostridium saccharide. The first two are strong fibrous degrading bacteria. The dominant bacteria in the gut of the pure blood horse are Clostridium pasteuriae, Clostridium amorinus, redox true bacilli, vivacious rumen coccus, Clostridium lactobacillus, Bifidobacterium lactate, Bacillus anthracis, Bacillus anthracis. Mongolia horse. The grazing conditions on the grassland were compared with the conditions of the captive breeding in March. The new dominant bacteria in the intestines were Vibrio acetate, true rectal bacilli, Clostridium Clostridium, unanimous cocci, and Bacillus subtilis. On the contrary, the Enterococcus aureus and Bacillus subtilis were not dominant bands in Mongolia horse grazing and grazing. The main fiber of the intestinal tract under two different feeding conditions in Mongolia horse The species of vitamin D changed little.

【学位授予单位】：内蒙古农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S821.81

【参考文献】