常见SPF级小鼠和大鼠肠道菌群多样性研究

Diversity of intestinal flora in commonly used SPF mice and rats

目的研究常见SPF级小鼠和大鼠的肠道菌群多样性。方法分别采集广东地区三家实验动物生产单位的C57BL/6、ICR、BALB/c小鼠和Wistar、SD大鼠的盲肠内容物样品,用细菌16S rDNA通用引物扩增V4-V5区域,采用Illumina Miseq 2×300 bp测序平台进行测序,使用生物信息学方法进行微生物群落分析、Alpha多样性分析与Beta多样性分析。结果对序列去杂优化后OTU聚类分析,稀释性曲线说明本次测序的数据量合理;实验小鼠和大鼠肠道菌群共分成八个门,其中拟杆菌门(Bacteroidetes)、厚壁菌门(Firmicutes)占据主要地位,属水平上主要是拟杆菌属(Bacteroides)、Hungatella、副杆状菌属(Parabacteroides)、乳酸杆菌属(Lactobacillus)等;样品间差异性分析显示相同设施来源动物的菌群组成相似性较高;Alpha分析结果显示来源于同种设施的动物物种丰富度相近;Beta分析显示相同设施动物的肠道菌群差异较小,但品系对肠道菌群差异性有所影响。结论不同来源设施的饲养环境是动物肠道菌群多样性的主要影响因素,不同品系对肠道菌群多样性有一定影响。

Objective To study the diversity of intestinal flora in commonly used SPF mice and rats. Methods The cecum contents of C57BL/6, ICR, BALB/c mice as well as Wistar and SD rats were collected from three experimental animal production units in the Guangdong area. The V4-V5 region was amplified using 16S rDNA primers and sequenced by the Illumina Miseq 2x 300 bp sequencing platform. Microbial community analysis, alpha diversity analysis, and beta diversity analysis were carried out by bioinformatics. Results In OTU cluster analysis after sequence removal optimization, the dilution curve indicated that the data quantity of the sequencing was reasonable. The intestinal microflora of mice and rats were divided into eight phyla. Bacteroidetes and Firmicute phyla occupied the main position, mainly in the level of Bacteroides, followed by the genuses Hungatella and Parabacteroides and then Lactobacillus. Analysis of the differences among the samples showed that the flora composition of animals from the same facility, and that of the same strains from the same facility was also similar. Alpha analysis showed that the species richness was similar in animals from the same facility, and beta analysis showed little difference in intestinal flora of the same origin animals. Moreover, the strain had an effect on the difference of intestinal flora. Conclusions The origin is the main factor influencing the diversity of intestinal flora, and the strain has a certain influence on their diversity.