16S rRNA基因高通量测序方法检测奶牛场常用干草表面微生物群落结构及多样性

Analysis of microbial community structure and diversity on surface of commonly used hay in dairy farm by 16S rRNA gene high-throughput sequencing

【目的】随着中国奶牛业的发展,干草需求量与日俱增。作为天然牧草,干草可以成为家畜传播病原体的载体。以干草表面附着物为研究对象,了解干草中细菌群落结构以及致病菌属特征。【方法】对来自6个不同奶牛场饲草舍的干草样本,应用Illumina Mi Seq高通量测序技术测定干草表面附着物细菌的16S r RNA基因V3-V4变异区序列,分析不同干草样本细菌群落组成。【结果】干草样本中的细菌在97%的相似水平下共得到OTU个数为15 416,涵盖了29门87纲144目219科323属的细菌。微生物多样性分析表明,干草样本具有很高的细菌多样性,不同样本多样性存在差异。对干草样本菌群中丰度较高的14种病原菌属进行分析,发现相较于人工种植牧草制备的干草,天然牧草制备的干草中病原菌属丰度较高。【结论】研究解析了干草样本中微生物的多样性、丰度及主要病原菌属的特征,对奶牛场疾病防控有一定指导意义。

[Objective] With the development of Chinese dairy industry, hay demand is increasing. As an natural forage, hay can be a carrier of livestock pathogens. The aim of this study was to investigate the bacterial community structure and the characteristics of pathogenic bacteria in hay. [Methods] We selected six different dairy farms to take hay samples. The Illumina Mi Seq sequencing with 16 S r RNA gene V3-V4 variable region was adopted to analyze the surface bacterial community structures of commonly used hay in dairy. [Results] A total number of 15 416 operational taxonomic units（OTUs） were obtained from hay samples under the similarity level of 97%, including 29 phyla, 87 classes, 144 orders, 219 families, and 323 genera. Microbial diversity analysis showed that hay samples had high bacterial diversity, and the diversity corresponding to each samples was specific. The results suggested that the abundance of pathogens in the natural hay was higher than that in the hay from artificial planted grasses. [Conclusion] The diversity, abundance, and characteristics of the main pathogens in hay samples were analyzed in this study, which would be a helpful reference for disease prevention and control in dairy farms.