基于高通量测序分析入境羊毛微生物群落结构特征

Microbial community characteristics of import wool revealed by high-throughput sequencing

目的通过高通量测序的方法了解来自5个国家5批入境羊毛表面携带的致病菌和微生物群落结构特征,为国境卫生以及传染病预防提供有效的数据支撑。方法通过浸泡搅拌震荡的方法收集5批羊毛样本表面的总细菌并提取DNA,采用Illumina Miseq高通量测序的方法,主要分析比较5批羊毛样本的α-多样性指数、微生物群落结构以及致病菌。结果α-多样性指数的结果表明5批羊毛表面微生物群落结构的多样性具有明显差异,其中德国样本的多样性最高,法国样本的多样性最低,美国、阿根廷、比利时则介于之间。主成分分析结果表明德国和比利时样本的微生物组成相似度较高且与其他3个国家之间的微生物群落结构组成有明显差异。微生物群落结构的16S绝对定量结果表明在微生物总量绝对丰度层面美国>阿根廷>德国>比利时>法国,阿根廷、德国、比利时3个国家的羊毛表面微生物以变形菌门(Proteobacteria)为主,厚壁菌门(Firmicutes)其次,而美国、法国以厚壁菌门(Firmicutes)为主,变形菌门(Proteobacteria)其次。在种属水平上,5批羊毛样本均检测出鲁氏不动杆菌(Acinetobacter lwoffii)、肠沙门菌(Salmonella enterica)和嗜麦芽窄食单胞菌(Stenotrophomonas maltophilia)等临床常见的条件致病菌,其中美国样本中肠沙门菌的丰度最高,比利时样本的肠沙门菌的丰度最低。结论根据测序的结果得出羊毛表面病原体含量较多且疫情复杂,检测出了肠沙门菌等危害性较大的致病菌,相关部门以及一线从业人员应注意做好生物防护。

To investigate the characteristics of pathogenic bacteria and microbial community of imported wool by high-throughput sequencing,and provide effective data support for border health and prevention of infectious diseases.The total bacteria on the surface of 5 batches of wool samples were collected by soaking,stirring and shaking,and then extracted by commercial kit.Alpha-diversity index,microbial community and pathogenic bacteria from five different countries were analyzed and compared by Illumina Miseq high-throughput sequencing.Theα-diversity index showed that the diversity of microbial community on the surface of five batches of wool had certain differences.Germany samples with the highest diversity,France samples with the lowest diversity and the diversity of United States,Argentina and Belgium between Germany and France were found.The principal component analysis showed that the microbial composition of samples from Germany and Belgium were similar but different from the other three countries.At the phylum level,Proteobacteria was the dominant phylum in Argentina,Germany and Belgium,followed by Firmicutes,in the USA and France,Firmicutes predominated,followed by Proteobacteria.Acinetobacter lwoffii,Salmonella enterica,Stenotrophomonas maltophilia and other clinically common opportunistic pathogens were detected at the species level,the American sample had the highest abundance of Salmonella enterobacter while Belgian sample had lowest abundance.These results indicated that there were many pathogens in wool and the epidemic situation was complicated.The detection of Salmonella enterica and other harmful bacteria reminds relevant departments and frontline employees to pay attention to biological protection.