中国媒介生物学及控制杂志 ›› 2023, Vol. 34 ›› Issue (3): 319-325.DOI: 10.11853/j.issn.1003.8280.2023.03.007

• 实验研究 • 上一篇    下一篇

贵州省少数民族自治州微小扇头蜱的宏基因组分析

向昱龙1, 周敬祝2, 张燕1, 胡勇1, 梁文琴1,2   

  1. 1. 贵州医科大学公共卫生与健康学院, 环境污染与疾病监控教育部重点实验室, 贵州 贵阳 550025;
    2. 贵州省疾病预防控制中心病媒生物监测科, 贵州 贵阳 550004
  • 收稿日期:2022-12-19 出版日期:2023-06-20 发布日期:2023-06-16
  • 通讯作者: 梁文琴,E-mail:liangwenqin521@126.com
  • 作者简介:向昱龙,男,在读硕士,主要从事蜱传病原体研究工作,E-mail:xiangyulong817@163.com
  • 基金资助:
    贵州省科技计划项目(黔科合支撑〔2022〕一般178);传染病预防控制国家重点实验室自主研究课题(2018SKLID305);贵州省传染病预防与控制人才基地科研团队-媒介生物及相关传染病监测预警中心(RCJD2107)

Metagenomic analysis of Rhipicephalus microplus from minority autonomous prefectures in Guizhou province, China

XIANG Yu-long1, ZHOU Jing-zhu2, ZHANG Yan1, HU Yong1, LIANG Wen-qin1,2   

  1. 1. School of Public Health, the Key Laboratory of Environmental Pollution Monitoring and Disease Control, Ministry of Education, Guizhou Medical University, Guiyang, Guizhou 550025, China;
    2. Vector Surveillance Section of Guizhou Center for Disease Control and Prevention, Guiyang, Guizhou 550004, China
  • Received:2022-12-19 Online:2023-06-20 Published:2023-06-16
  • Supported by:
    Plan Project of the Science and Technology in Guizhou Province (Qian Ke He Support [2022] General 178); Independent Research Project of the State Key Laboratory of Infectious Disease Prevention and Control (No. 2018SKLID305); Scientific Research Team of Guizhou Provincial Infectious Disease Prevention and Control Talent Base, Center for Surveillance and Early Warning of Vectorial Organisms and Related Infectious Diseases (No. RCJD2107)

摘要: 目的 调查贵州省少数民族自治州的优势寄生蜱种微小扇头蜱的微生物菌群,为蜱传疾病防控提供科学依据。方法 2019-2020年4和7月在黔东南苗族侗族自治州、黔南布依族苗族自治州和黔西南布依族苗族自治州的18个采样点捕捉牛、羊体表的微小扇头蜱。将捕获的微小扇头蜱按地区、生长阶段和性别分组,每组3个平行样,进行宏基因组测序。测序结果经质控、拼接组装等操作后与美国国立生物技术信息中心(NCBI)非冗余蛋白库进行同源性比对,获得物种分类注释信息。然后通过R(v. 3.6.3)和GraPhlAn(v. 1.1.3)软件进行可视化分析,包括物种组成分析、非度量多维尺度(NMDS)分析和组间相似性分析(ANOSIM)。结果 微小扇头蜱的优势菌门为厚壁菌门(60.70%),其次为变形菌门(33.76%)和放线菌门(4.53%)。优势菌种为金黄色葡萄球菌(38.29%),其次是肺炎克雷伯菌(7.79%)和肺炎链球菌(5.26%)。检出多种蜱传病原体,包括斑点热群立克次体的多种基因型和嗜吞噬细胞无形体(1.38%)。Rickettsia fournieri(2.91%)是中国首次从蜱中检出的新发立克次体,日本立克次体(1.76%)和R. monacensis(0.04%)是中国常见的蜱传立克次体基因型。R. fournieri和日本立克次体主要由雄蜱携带。NMDS分析表明,不同生长阶段、不同性别微小扇头蜱的微生物菌群组成不同。ANOSIM分析结果显示分组合理,组间差异大于组内差异(R=0.147,P=0.014)。结论 贵州省少数民族自治州的微小扇头蜱微生物菌群组成丰富,携带的病原体可能会导致斑点热和人粒细胞无形体病,应加强相关媒介监测和病原体检测,采取相关防控措施,降低蜱传疾病发病风险。

关键词: 微小扇头蜱, 宏基因组分析, 细菌, 斑点热群立克次体, 嗜吞噬细胞无形体, 贵州省

Abstract: Objective To investigate the microbiota of the dominant tick species Rhipicephalus microplus in the minority autonomous prefectures of Guizhou province, China, and to provide a scientific basis for the prevention and control of tick-borne disease.Methods Rh. microplus ticks were collected from the body surface of cattle and sheep at 18 sampling sites located in Qiandongnan Miao and Dong Autonomous Prefecture, Qiannan Buyi and Miao Autonomous Prefecture, and Qianxinan Buyi and Miao Autonomous Prefecture in April and July of 2019 and 2020. The ticks were grouped by region, growth stage, and sex, with three parallel samples in each group, followed by metagenomic sequencing. The sequencing results were processed with quality control and assembly procedures before homology alignment with the non-redundant protein database of National Center for Biotechnology Information to obtain taxonomic annotation information. R (v. 3.6.3) and GraPhlAn (v. 1.1.3) were used for visual analysis, including species composition analysis, non-metric multidimensional scaling (NMDS) analysis, and analysis of similarities (ANOSIM).Results The dominant bacterial phylum for Rh. microplus was Firmicutes (60.70%), followed by Proteobacteria (33.76%) and Actinobacteria (4.53%). Staphylococcus aureus (38.29%) was the dominant bacterial species, followed by Klebsiella pneumoniae (7.79%) and Streptococcus pneumoniae (5.26%). Several tick-borne pathogens were detected, including various genotypes of spotted fever group Rickettsia and Anaplasma phagocytophilum (1.38%). R. fournieri (2.91%) was an emerging rickettsiae detected from ticks for the first time in China, and R. japonica (1.76%) and R. monacensis (0.04%) were common tick-borne Rickettsia genotypes in China. R. fournieri and R. japonica were mainly carried by male ticks. The NMDS analysis showed that the microbiota composition of Rh. microplus differed by growth stage and sex. The ANOSIM analysis indicated reasonable grouping, with greater intergroup differences than intragroup differences (R=0.147, P=0.014).Conclusions In the minority autonomous prefectures of Guizhou province, Rh. microplus ticks have a diverse microbiota composition, carrying pathogens that may cause spotted fever and human granulocytic anaplasmosis. Therefore, vector surveillance and pathogen detection should be strengthened, and effective control measures should be taken to reduce the risk of tick-borne diseases.

Key words: Rhipicephalus microplus, Metagenomic analysis, Bacteria, Spotted fever group Rickettsia, Anaplasma phagocytophilum, Guizhou province

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