不同野生动物肠道微生物多样性分析及功能初步研究

发布时间：2018-06-23 05:36

本文选题：野生动物 + 肠道微生物　；参考：《吉林农业大学》2015年硕士论文

【摘要】：目前,国内外研究人员利用现代分子生物学技术在动物肠道微生物研究领域取得诸多进展,如确立了宿主与微生物之间的相互关系,提高了宿主的健康状态和营养水平,增加了动物对饲料的利用效率,明确了胃肠道微生物的代谢通路等。近年来对野生动物肠道微生态的研究得到广泛关注,研究野生动物肠道微生物不仅可以描述野生动物肠道微生物的系统发育,分析其肠道菌群功能,研究其胃肠道微生物区系变化规律、信号通路及其响应机制,而且可以通过改变肠道微生物组成、增加优势菌群数量等有效措施预防和治疗某些肠道疾病,进而为野生动物健康可持续的繁殖和生存做出贡献。本研究采用PCR-DGGE技术研究不同野生动物肠道菌群多样性,经分析得到23条差异条带,测序结果表明,厚壁菌门(Firmicutes)为肠道优势菌群。对草食动物与肉食动物肠道菌群分析可知,芽孢杆菌科(Bacillaceae)的梭菌属(Fusobacterium)在草食动物肠道菌群的属水平占优势地位。在反刍动物粪便中检测出具有高效纤维分解能力的溶纤维丁酸弧菌(Butyrivibrio fibrisolvens)。消化链球菌属(Peptostreptococcus)为肉食动物肠道菌群的优势菌属,在犬科肉食动物粪便中检测到Anaerovorax,在猫科动物粪便中分离出特有菌为河流漫游球菌(Vagococcus fluvialis)。本研究同时选择宏基因组测序技术研究野生动物肠道菌群多样性,共得到242 609条序列,40 130个操作分类单元(OTUs),其中在草食动物肠道菌群中占优势地位的细菌归类为Firmicutes、拟杆菌门(Bacteroidetes)、变形菌门(Proteobacteria)、放线菌门(Actinobacteria)和疣微菌门(Verrucomicrobia),肉食动物肠道菌群中优势菌门为Firmicutes、Actinobacteria、Proteobacteria、Bacteroidetes和梭杆菌门(Fusobacteria),在属水平上,草食动物肠道微生物明显比肉食动物肠道微生物种类丰富,在单胃草食动物和反刍动物肠道菌群比较中,具有明显差异的菌属为瘤胃球菌属(Ruminococcus)、颤杆菌克(Oscillibacter)、鸟杆菌属(Ornithobacterium)、Anaerovorax、Paludibacter、Alistipes和Succinivibrio。在肉食动物中,犬科动物和猫科动物肠道微生物多样性表现出差异的菌属为醋弧菌属(Acetivibrio)、假单胞菌属(Pseudomonas)、Sporobacter、Bulleidia和Eggerthella。在现代微生物多样性研究方法基础上,本研究进一步利用传统分离培养方法对野生动物粪便中菌群进行分离、鉴定,然后对菌株的纤维素酶、木质素酶、蛋白酶、淀粉酶及脂肪酶相关功能进行研究,同时根据菌株功能对肠道菌群功能进行分析,并对PCR-DGGE技术和宏基因组测序技术所得结果进行验证,发现利用宏基因组测序技术得到的结果较PCR-DGGE技术更加全面;并确定草食动物与肉食动物粪便中存在相同菌株,它们是Bacillus drentensis和Bacillus safensis,发现其具有产纤维素酶、蛋白酶和淀粉酶功能;分析菌群多样性与菌株功能之间关系时发现,从不同食性动物粪便中分离出的菌株在功能上有所差异,从草食动物粪便中分离出的菌株多具有纤维素分解功能,而从肉食动物粪便中分离的菌株多具有脂肪分解功能。通过以上研究,所得结果能够初步反映不同野生动物肠道菌群结构及其功能特征,这为今后野生动物肠道菌群多样性的研究以及功能优势菌株的筛选提供一定理论依据,并为野生动物消化功能相关基因研究以及野生动物健康繁殖与生存奠定一定基础。
[Abstract]:At present, researchers at home and abroad have made great progress in the field of animal intestinal microflora by modern molecular biology technology, such as establishing the relationship between host and microorganism, improving the health and nutrition level of the host, increasing the utilization efficiency of animal feed and clarified the metabolic pathways of gastrointestinal microorganism. In recent years, the research on the intestinal microflora of wild animals has been widely concerned. The study on the intestinal microorganism of wild animals can not only describe the phylogenetic development of the intestinal microbes in wild animals, analyze the intestinal microflora function, study the changes in the microbiological flora of the gastrointestinal tract, the signaling pathway and its response mechanism, and can also change the intestinal tract by changing the intestinal tract. Microbial composition, increasing the number of dominant bacteria groups, and other effective measures to prevent and treat certain intestinal diseases, and thus contribute to the healthy and sustainable reproduction and survival of the wild animals. This study used PCR-DGGE technology to study the diversity of intestinal flora in different wild animals. 23 different bands were analyzed, and the results of sequencing showed that Fir Micutes) for the intestinal predominant flora. The analysis of the intestinal flora of herbivorous and carnivorous animals showed that the Fusobacterium of Bacillaceae was dominant in the level of intestinal flora of herbivorous animals. In ruminant feces, the soluble Vibrio butyrate (Butyrivibrio FIB with high efficiency of fibrinolysis) was detected. Risolvens. Peptostreptococcus is the dominant bacteria in the intestinal flora of carnivorous animals. Anaerovorax is detected in the faeces of canine carnivorous animals, and the endemic bacteria isolated from the faeces of the cat family is Vagococcus fluvialis (Vagococcus fluvialis). This study selected the macrogenome sequencing technology to study the intestines of wild animals in the same time. There were 242609 sequences and 40130 operating taxa (OTUs), among which the dominant bacteria in the herbivorous intestinal flora were classified as Firmicutes, Bacteroidetes, Proteobacteria, Actinobacteria and Verrucomicrobia, and the intestinal flora of carnivorous animals The dominant bacteria were Firmicutes, Actinobacteria, Proteobacteria, Bacteroidetes and Clostridium (Fusobacteria). At the level of the genus, the intestinal microbes of the herbivorous animals were obviously more abundant than those of the carnivorous animals. In the comparison of the intestinal flora of the single stomach herbivore and the ruminant, the bacteria with the obvious difference were the rumen coccus (R). Uminococcus), Oscillibacter, Ornithobacterium, Anaerovorax, Paludibacter, Alistipes and Succinivibrio. in carnivorous animals. The diversity of intestinal microbial diversity in canine and feline intestinal microflora is a genus of Vibrio (Acetivibrio), Pseudomonas (Pseudomonas), Sporobacter, Bulleidia and Eggerthella. on the basis of modern microbial diversity research methods, this study further uses the traditional isolation and culture methods to isolate and identify the bacteria in the faeces of wild animals, and then studies the related work of cellulase, lignin, protease, amylase and lipase, and the function of the strains to the intestinal flora. The function is analyzed, and the results obtained by PCR-DGGE technology and macrogenome sequencing technology are verified. It is found that the results obtained by the macro genome sequencing technology are more comprehensive than the PCR-DGGE technology, and determine the same strains in the faeces of herbivorous animals and carnivorous animals, they are Bacillus drentensis and Bacillus safensis. With the function of producing cellulase, protease and amylase, the analysis of the relationship between the diversity of the flora and the function of the strain found that the strains isolated from the faeces of different food animals were different in function, and most of the strains isolated from the faeces of herbivorous faeces had the power of cellulose decomposition, and the strains isolated from the faeces of carnivorous animals were many. The results can preliminarily reflect the structure and functional characteristics of intestinal microflora in different wild animals. This provides a theoretical basis for the study of the diversity of intestinal flora and the screening of functional dominant strains in the future, and the study of the genes related to the digestive function of wild animals and the wild animals. It lays a foundation for the healthy reproduction and survival of animals.
【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】：S852.6

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