野生和圈养川金丝猴肠道菌群季节动态及功能研究

Golden snub-nosed monkey is a herbivorous animal, has vertical migration in its habitat with the season changes. Gut microbiota assists in intestinal homeostasis and other aspects of the host, including intestinal immune response, digestion, and physiology. A remarkable microbiota was obtained from the feces of golden snub-nosed monkey. In addition, the composition and diversity were closely related to the health of host. However, the research of gut microbiota in wild golden snub-nosed monkey is rare, particularly in the influence of seasonal variation. This could be due to the limitations of the animal studied and research methods. This study aimed to investigate the bacterial communities in the feces of wild and captive golden snub-nosed monkey in different seasons. First, we undertake an analysis of DGGE and real-time PCR to study the composition and diversity of gut microbiota. Second, we used a high-throughput sequencing technology to analyze the intestinal flora. This could help us to obtain a preliminary analysis of the gut microbiota. Finally, we utilize a combination of metagenomics and metabolomics to investigate the mechanism between gut microbiota and metabolites. Our findings provide important basic information for the conservation and management of golden snub-nosed monkey, particularly in the practical guidance of the diet composition in captivity monkey.

川金丝猴是植食性动物，随季节的变化在栖息地中作垂直迁徙。环境可影响宿主肠道菌群的结构和功能，进而影响宿主的健康。本课题组以往的研究表明圈养川金丝猴肠道中栖息着大量而丰富的菌群，且菌群组成及多样性与金丝猴的健康紧密相关。但少见报道野生川金丝猴肠道菌群，特别是未见报道季节变化对野生和圈养川金丝猴肠道菌群及功能的影响，可能是源于研究对象以及研究方法的局限。本项目以不同季节野生和圈养川金丝猴肠道菌群为研究对象，首先应用变性梯度凝胶电泳技术和实时荧光定量PCR技术分析菌群组成及多样性；其次，利用高通量测序技术大规模分析川金丝猴肠道菌群，初步分析关键菌群；最后，应用宏基因组学和代谢组学技术联合分析肠道关键菌群与关键代谢物相关机制，挖掘川金丝猴肠道关键菌群与代谢物，为保护和管理川金丝猴种群提供重要的基础资料，特别是为圈养川金丝猴的日粮组成提供现实指导。

肠道共生微生物与宿主的免疫应答、消化和生理密切相关。研究野生动物肠道微生物对其物种的保护具有重要作用。川金丝猴是濒危犹猴亚科植食性灵长类，本项目以川金丝猴为研究对象，运用变性梯度凝胶电泳技术、实时荧光定量PCR技术、高通量测序等技术，分析不同季节野生和圈养川金丝猴肠道菌群及功能。得到以下结果：1、季节变化和生存环境（野生和圈养）影响川金丝猴肠道菌群，厚壁菌门（Firmicutes，65.18 %）、拟杆菌门（Bacteroidetes，18.08 %）和（Proteobacteria，3.75 %）是川金丝猴的优势菌群，厚壁菌门在秋季野外样品中丰度较高；2、代谢活动主要是氨基糖和核苷酸糖代谢，与该代谢活动相关的酶主要是α-L-阿拉伯基呋喃糖苷酶（EC 3.2.1.55），其参与降解阿拉伯聚糖。并且，该酶与川金丝猴肠道优势的糖苷水解酶类GH2、GH3和GH43有关；3、与α-L-阿拉伯基呋喃糖苷酶相关的菌群主要来自厚壁菌门，分别是Firmicutes bacterium CAG:110和Faecalibacterium prausnitzii，其丰度在野外粪便样品中较高；4、与该酶相关的菌群中，Faecalibacterium sp. CAG:74, Roseburia hominis 和 uncultured Clostridium sp.是野生样品特有；而Roseburia sp. CAG:100、uncultured Ruminococcus sp.、Bacteroides sp. CAG:709 和Prevotella sp. CAG:873是圈养样品特有的菌群，且这些菌群主要来自拟杆菌门，说明拟杆菌门在圈养动物肠道消化方面同样发挥着重要作用。由此可见，野外粪便样品菌群具有较高的肠道菌群代谢活动，并与厚壁菌门菌群密切相关，推测与动物的饮食差异有关。 本研究证实季节因素及生存环境均影响川金丝猴肠道菌群及功能；同时还发现川金丝猴肠道菌群仍有大量未知功能基因。总的来说，本研究挖掘到与季节变化和生存环境相关的川金丝猴肠道关键菌群及功能，丰富了该濒危物种的肠道菌群及功能信息，并为圈养野生动物提供现实指导。