本文選題：蒙古馬 + 腸道細菌��； 參考：《內(nèi)蒙古農(nóng)業(yè)大學》2015年碩士論文

【摘要】：蒙古馬是我國著名的地方品種,有著優(yōu)秀的品種遺傳資源。已被錄入我國保種名錄中。比其它馬種蒙古馬具有耐粗飼特性,抓膘迅速,掉膘緩慢。這也是蒙古馬長期在惡劣環(huán)境條件下進化出來的生存能力。蒙古馬的耐粗飼特性除了遺傳因素,必然與其特殊的腸道微生物區(qū)系相關(guān)。為了了解其中微生物作用關(guān)系,本實驗對蒙古馬和純血馬腸道細菌多樣性初步研究。也為功能細菌的挖掘和利用提供基礎(chǔ)。本實驗以純血馬和不同飼養(yǎng)條件下的蒙古馬新鮮糞便為實驗樣品,用生物試劑盒直接從糞樣中提取細菌基因組DNA,使用PCR技術(shù)從細菌總DNA中擴增出16SrDNA V3區(qū)序列并采用變性梯度凝膠電泳分離不同細菌的基因序列。用Quantity One軟件對膠圖進行分析,同時切膠回收優(yōu)勢條帶進行克隆測序。將得到的測序在基因庫數(shù)據(jù)里進行Blast比對判斷菌種屬性。實驗主要結(jié)果以下：第一、蒙古馬和純血馬腸道細菌群落結(jié)構(gòu)比較中,DGGE膠圖顯示不同位置的條帶有53個。選擇41個優(yōu)勢條帶回收測序鑒定中與NCBI數(shù)據(jù)庫細菌16S rDNAV3區(qū)序列相似性96%以上的僅有16個克隆。在蒙古馬兩種不同飼養(yǎng)條件下的腸道細菌結(jié)構(gòu)變化的研究中,DGGE膠圖中顯示共有50個條帶,選40個優(yōu)勢條帶回收測序,結(jié)果序列相似性96%以上有17個克隆。因此同源性較低。兩次實驗中分別未報道的優(yōu)勢細菌占總細菌的約61%和58%。第二、利用Quantity One軟件分析出的DGGE圖譜識別的條帶數(shù)目和群落多樣性指標來看,純血馬腸道優(yōu)勢菌比蒙古馬腸道優(yōu)勢菌更豐富多樣。聚類分析結(jié)果蒙古馬和純血馬各自聚成一支,品種之間腸道優(yōu)勢菌相似性僅為0.33。品種內(nèi)相似性都大于品種間相似性。蒙古馬7月份草原上放牧時的腸道優(yōu)勢菌比3月份圈養(yǎng)時的腸道優(yōu)勢菌更豐富多樣。聚類分析后蒙古馬3月份和7月份各自聚成一支,之間相似性為0.54。蒙古馬吃干草情況下的種內(nèi)相似性大于吃青草時的。第三、蒙古馬和純血馬腸道內(nèi)共有的優(yōu)勢菌有棲瘤胃普雷沃氏菌、溶纖維丁酸弧菌、微黃棒狀桿菌、黃化瘤胃球菌和枯草芽孢桿菌,以上菌種中除了枯草芽孢桿菌都具有降解纖維功能�？莶菅挎邨U菌是多功能益生菌。蒙古馬腸道內(nèi)特有的優(yōu)勢菌有瘤胃球菌屬、解纖維梭菌、化糖梭狀芽胞桿菌。其中前兩者是較強的纖維降解菌。純血馬腸道內(nèi)特有的優(yōu)勢菌有巴氏梭狀芽胞桿菌、嗜胺梭菌、氧化還原真桿菌、活潑瘤胃球菌、共生梭菌、乳酸雙歧桿菌、牛鏈球菌、炭疽芽孢桿菌。蒙古馬7月份草原上放牧比3月份圈養(yǎng)條件,腸道內(nèi)新增優(yōu)勢菌有解纖維醋弧菌、直腸真桿菌、梭狀梭菌、一致糞球菌、短真桿菌。相反微黃棒狀桿菌和枯草芽孢桿菌在蒙古馬放牧吃青草情況下變成不是優(yōu)勢條帶。蒙古馬兩種不同飼養(yǎng)條件下腸道主要纖維菌的種類變化不大。
[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.

【學位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S821.81

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