革蜱携带微生物多样性的二代测序技术分析  

Analysis of microbial diversity carried by Dermacentor silvarum through next-generation sequencing

作　　者：宋春玲 胡阳 唐亮 李倩 郭鑫宇 柏慧[1] 刘慧婷[1] 王春仁[1] 常巧呈[1] SONG Chunling;HU Yang;TANG Liang;LI Qian;GUO Xinyu;BAI Hui;LIU Huiting;WANG Chunren;CHANG Qiaocheng(College of Animal Science and Veterinary Medicine,Heilongjiang Bayi Agricultural University,Daqing 163319,China)

机构地区：[1]黑龙江八一农垦大学动物科技学院

出　　处：《黑龙江畜牧兽医》2018年第21期5-9,254,共6页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金青年基金项目(31302080);国家级大学生创新创业训练计划项目(201710223008);黑龙江八一农垦大学“青年创新人才”项目(CXRC2016-08)

摘　　要：为了解黑龙江省尚志市某林区革蜱携带微生物种类的特点,试验采集游离蜱,随机取雌雄革蜱各96只,提取蜱足和其他部位的DNA样品分成雌蜱、雄蜱和蜱足3组,分别扩增16S r DNA基因V3-V4区,应用Illumina Hi Seq 2500测序平台进行深度测序,并对微生物多样性进行生物信息学分析。蜱足组代表外环境微生物群,在去除蜱足组所携带微生物的有效序列后,对雌蜱和雄蜱两组革蜱体内的微生物进行具体分析。结果表明：雌蜱和雄蜱两组样品共获得60 954条有效序列,平均序列长度为423 bp,包括651个分类操作单元（OTU）,共192个科,309个属;其中检测到25类致病微生物,分别包括布鲁氏菌科、巴斯德氏菌科和分支杆菌属等常见人兽共患病菌,以及考克斯氏体属、立克次氏体属、无形体属、疏螺旋体属和埃立克体属5类蜱传病原;分析发现雌蜱和雄蜱携带的病原种类和丰度均不同,其中雌蜱中蜱传病原的种类多于雄蜱。说明采自尚志市某林区森林革蜱体内的微生物群结构具有多样性和复杂性,雌蜱和雄蜱微生物群的多态性存在差异。The aim of the present study was to investigate species characteritics of bacteria carried by Dermacentor sivarum from a forest district of Shangzhi City in Heilongjiang Province. Free ticks were collected, and 96 female and 96 male Dermacentor sivarurn were randomly selected. The DNA samples of tick foot and other parts were extracted and divided into three groups ： female tick, male tick and tick foot. The V3 - V4 region of 16S rDNA gene was amplified respectively. The IUumina HiSeq 2500 sequencing platform was used for in-depth sequencing, and the micro- bial diversity was analyzed by bioinformatics. After removing the microbial effective sequences of the bacteria carried by the tick foot group, the diversity in the female tick and male tick groups were analyzed. The results showed that 60 954 effective sequences were obtained in female tick and male tick groups, with an average sequence length of 423 bp, including 651 OTUs, including 192 families and 309 genera. Among them, 25 kinds of pathogenic microorganisms were detected, including Brucellaceae, PasteureUaceae, Mycobacterium and other common zoonotie bacte- ria, and including 5 kinds of tick-borne pathogens such as Coxiella, Rickettsia, Anaplasma, Borrelia and Ehrlichia. It was found that the spe- cies and abundance of pathogens carried by female ticks and male ticks were different, and the species of tick-borne pathogens in female ticks were more than that in male ticks. The results indicated that the microbial community structure of Dermacentor silvarum collected from a forest area of Shangzhi City was diverse and complex, and there was a difference in the polymorphism between female and male ticks.

关 键 词：黑龙江省 革蜱 二代测序 微生物多样性 人兽共患病菌 蜱传病原 

分 类 号：S855.99[农业科学—临床兽医学]