基于高通量测序技术分析人工圈养成年蜂猴的粪便微生物多样性

Analysis of Microbial Diversity in Feces of Adult Nycticebus bengalensis by High-throughput Sequencing

目的研究成年蜂猴的肠道菌群组成情况,以探讨人工圈养蜂猴的健康影响因素。方法采集4只成年雄性蜂猴的粪便样本,运用细菌16SrDNA通用引物扩增V4区,应用IlluminaNovaSeq测序平台对蜂猴粪便微生物进行测序分析,并使用Qiime软件分析样本的复杂度和相似度。基于有效数据分析样本肠道菌群在门、属水平上的物种结构和丰度,同时应用PICRUSt软件进行菌群的代谢功能预测。结果年龄最小的蜂猴的肠道菌群多样性指数shannon高于其他蜂猴。PCoA分析显示4只蜂猴的样本菌群组成具有一定的相似度。4只蜂猴的粪便中细菌共鉴定出了12个门、18个纲、28个目、49个科、93个属和59个种。其中,优势菌门是拟杆菌门(Bacteroidetes)和厚壁菌门(Firmicutes),平均相对丰度分别为46%和28%;拟杆菌属(Bacteroides)、双歧杆菌属(Bifidobacterium)和梭菌属(Fusobacterium)的丰度较高,平均相对丰度分别为33%、6%和6%;有益菌群双歧杆菌属在4个蜂猴粪便样本中都存在,且在年龄较小的蜂猴中相对丰度最高。PICRUSt功能预测分析显示,蜂猴粪便菌群中氨基酸转运和代谢、碳水化合物转运和代谢等功能基因的丰度相对较高。结论应用IlluminaNovaSeq高通量测序技术可以较全面地检测蜂猴的粪便菌群。蜂猴粪便中细菌组成具有丰富的多样性,其中还有许多未被分类鉴定且相对丰度较高的细菌,需要进一步研究。

Objective To analyze the composition of microbial community in gut of elderly male Nycticebusbengalensis,aiming to explore the health impact factors associated with artificial captivity.Methods Fecal samples of 4 adult male Nycticebusbengalensiswere collected and the V4 region was amplified using bacterial 16S rDNA universal primers.Illumina NovaSeq sequencing platform was used for microbial sequencing analysis.The complexity and similarity of the samples were analyzed using the QIIME software analysis tool.The species structure and abundance of the intestinal flora were analyzed at the phylum and genus levels based on validated data.The PICRUSt software was applied to predict the metabolic function of the flora.Results The results showed that the diversity index(Shannon index)of the flora in the youngest Nycticebusbengalensiswas higher than that of the others.PCoA analysis showed that the bacterial community compositions of the four samples had a certain degree of similarity.Bacteria identified in the Nycticebusbengalensis feces included 12 phyla,18 classes,28 orders,49 families,93 genera,and 59 species.Among them,the dominant phyla were Bacteroidetes and Firmicutes with average relative abundances of 46%and 28%,respectively;The genera Bacteroides,Bifidobacterium,and Fusobacteriumhad higher abundances,with relative abundances of 33%,6%,and 6%,respectively;The beneficial genus Bifidobacteriumwas found in all samples,with the highest relative abundance found in the younger Nycticebusbengalensis.PICRUSt function prediction analysis showed that the abundance of the functional genes related to amino acid transport and metabolism,carbohydrate transport and metabolism,was relatively high.Conclusion The use of Illumina NovaSeq high-throughput sequencing technology can comprehensively detect the fecal microbial community of the Nycticebusbengalensis.The bacterial composition of Nycticebusbengalensishas rich diversity.Many bacteria are not identified and have higher relative abundance,which need further study.