贵州省3个自治州寄生蜱细菌群落多样性研究

doi:10.12140/j.issn.1000-7423.2025.03.011

中国寄生虫学与寄生虫病杂志 ›› 2025, Vol. 43 ›› Issue (3): 370-376.doi: 10.12140/j.issn.1000-7423.2025.03.011

贵州省3个自治州寄生蜱细菌群落多样性研究

管毓威¹(), 向昱龙², 周敬祝³, 罗小龙³, 孔雪雪¹, 张燕¹, 胡勇¹, 梁文琴¹^,³^,^*()

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 561113
2 沙坪坝区疾病预防控制中心，重庆 400038
3 贵州省疾病预防控制中心病媒生物监测科，贵州省微生物组与传染性疾病防控重点实验室，贵州贵阳 550004

收稿日期:2024-11-14 修回日期:2025-02-02 出版日期:2025-06-30 发布日期:2025-05-21
通讯作者: *梁文琴，女，博士，主任技师，从事媒介生物及相关传染病防控研究。E-mail：liangwenqin521@126.com
作者简介:管毓威，男，硕士研究生，从事蜱媒病原体研究。E-mail：2456699183@qq.com
基金资助:
贵州省科技计划(Qiankehe support[2022]general 178);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);贵州省微生物组与传染性疾病防控重点实验室(ZDSYS[2023]004)

Investigation of bacterial community diversity in parasitic ticks from three autonomous prefectures in Guizhou Province

GUAN Yuwei¹(), XIANG Yulong², ZHOU Jingzhu³, LUO Xiaolong³, KONG Xuexue¹, ZHANG Yan¹, HU Yong¹, LIANG Wenqin¹^,³^,^*()

1 School of Public Health, Key Laboratory of Environmental Pollution Monitoring and Disease Control, Ministry of Education, Guizhou Medical University, Guiyang 561113, Guizhou, China
2 Shapingba District Center for Disease Control and Prevention, Chongqing 400038, China
3 Key Laboratory of Microbio and Infectious Disease Prevention & Control in Guizhou Province, Vector Surveillance Section of the Guizhou Center for Disease Control and Prevention, Guiyang 550004, Guizhou, China

Received:2024-11-14 Revised:2025-02-02 Online:2025-06-30 Published:2025-05-21
Contact: *E-mail: liangwenqin521@126.com
Supported by:
Guizhou Provincial Science and Technology Plan Project(Qiankehe support[2022]general 178);Scientific Research Team of the Infectious Disease Prevention and Control Talent Base in Guizhou Province-Vector Biology and Related Infectious Disease Monitoring and Early Warning Center(RCJD2107);Guizhou Provincial Key Laboratory of Microbiology and Infectious Disease Prevention and Control(ZDSYS[2023]004)

摘要/Abstract

摘要：

目的了解贵州省3个自治州内蜱的种类和蜱携带微生物群的多样性。方法 2019年4月和2020年7月，在贵州省黔东南苗族侗族自治州（以下简称黔东南）、黔南布依族苗族自治州（以下简称黔南）和黔西南布依族苗族自治州（以下简称黔西南）采集牛、羊和鼠体表的寄生蜱并进行形态学鉴定。按采集地区和蜱种将蜱分成6组，提取DNA后进行16S rDNA高通量测序。测序结果经操作分类单元（OTU）聚类分析后与核糖体数据库进行比对获得物种注释，进行细菌群落组成分析和α多样性分析。根据β多样性距离矩阵进行层级聚类，采用布雷柯蒂斯算法构建样品聚类树。采用R 3.6.3软件进行非度量多维尺度（NMDS）和组间相似性分析。结果共采集蜱1 463只，包括1 227只微小扇头蜱（83.87%）、208只长角血蜱（14.22%）、25只粒形硬蜱（1.71%）和3只褐黄血蜱（0.20%）。在黔东南、黔南和黔西南分别采集蜱220、1 039和204只，占采集总数的15.04%、71.02%、13.94%。高通量测序共产生1 682个OTU。α多样性分析显示，黔东南长角血蜱的香农指数、Chao1指数、Ace指数和均匀度指数均较高，分别为4.868、568.481、567.479和0.770。细菌群落鉴定共注释到29个菌门、70个菌纲、118个菌目、245个菌科和503个菌属，变形菌门、α-变形菌纲、立克次体目、立克次体科和立克次体属分别为各分类水平的优势菌，平均相对丰度分别为75.52%、57.70%、52.47%、50.88%和50.88%。微小扇头蜱携带的菌属以立克次体属（89.45%）和柯克斯体属（2.82%）为主，长角血蜱以立克次体属（18.06%）和假单胞菌属（11.98%）为主，粒形硬蜱以螺原体属（33.19%）和葡萄球菌属（22.22%）为主。黔东南、黔南和黔西南蜱携带的优势菌属均为立克次体属，平均相对丰度分别46.96%、60.20%和45.47%。样品聚类树显示，3个地区的微小扇头蜱样品与1份黔南的长角血蜱样品聚为一支，余下的长角血蜱样品聚为一支，黔西南的3份粒形硬蜱样品聚为一支。NMDS和组间相似性分析结果显示，6组样品的细菌群落组成不同（R = 0.599，P < 0.01）。结论贵州省3个自治州的蜱携细菌群落组成丰富，立克次体属为优势菌属。不同蜱种的细菌群落组成不同。

关键词: 微小扇头蜱, 长角血蜱, 粒形硬蜱, 细菌群落多样性, 高通量测序

Abstract:

Objective To investigate the species and microbial diversity of ticks from three autonomous prefectures in Guizhou Province. Methods In April 2019 and July 2020, parasitic ticks on the body surfaces of cattle, sheep and rodents were collected in Qiandongnan Miao and Dong Autonomous Prefecture (referred to as Qiandongnan), Qiannan Buyi and Miao Autonomous Prefecture (referred to as Qiannan) and Qianxinan Buyi and Miao Autonomous Prefecture (referred to as Qianxinan) in Guizhou Province, followed by morphological identification. The ticks were divided into 6 groups based on the collection regions and tick species. After DNA extraction, 16S rDNA high-throughput sequencing was performed. The sequencing results were analyzed by operational taxonomic unit (OTU) classification cluster and compared with the ribosomal database to obtain species annotations, enabling bacterial community composition analysis and α diversity analysis. Hierarchical clustering was conducted based on the β diversity distance matrix and a sample clustering tree was constructed using the Bray-Curtis algorithm. Non-metric multidimensional scale (NMDS) analysis and inter-group similarity analysis were performed using R 3.6.3 software. Results A total of 1 463 ticks were collected, including 1 227 Rhipicephalus microplus (83.87%), 208 Haemaphysalis longicornis (14.22%), 25 Ixodes granulatus (1.71%) and 3 Haemaphysalis flava (0.20%). In Qiandongnan, Qiannan and Qianxinan, 220, 1 039 and 204 ticks were collected respectively, accounting for 15.04%, 71.02% and 13.94%. A total of 1 682 OTUs were generated. α diversity analysis revealed that H. longicornis from Qiandongnan exhibited relatively high Shannon index, Chao1 index, Ace index and evenness index values of 4.868, 568.481, 567.479 and 0.770, respectively. Bacterial community identification annotated a total of 29 phyla, 70 classes, 118 orders, 245 families and 503 genera. Proteobacteria, Alphaproteobacteria, Rickettsiales, Rickettsiaceae and Rickettsia emerged as the dominant taxa at their respective taxonomic levels, with average relative abundances of 75.52%, 57.70%, 52.47%, 50.88% and 50.88%, respectively. The bacterial genera carried by R. microplus were predominantly Rickettsia (89.45%) and Coxiella (2.82%), while H. longicornis mainly harbored Rickettsia (18.06%) and Pseudomonas (11.98%), and I. granulatus primarily carried Spiroplasma (33.19%) and Staphylococcus (22.22%). The dominant bacterial genus in ticks from Qiandongnan, Qiannan, and Qianxinan was Rickettsia, with average relative abundances of 46.96%, 60.20% and 45.47%, respectively. The sample clustering tree demonstrated that R. microplus samples from 3 regions clustered together with a H. longicornis sample from Qiannan, while the remaining H. longicornis samples formed a separate cluster, and 3 I. granulatus samples from Qianxinan clustered independently. NMDS and inter-group similarity analysis indicated distinct bacterial community compositions among the 6 sample groups (R = 0.599, P < 0.01). Conclusion The bacterial communities carried by ticks in the three autonomous prefectures of Guizhou Province exhibited rich diversity, with Rickettsia being the dominant genus. Additionally, the bacterial community compositions differed among various tick species.

Key words: Rhipicephalus microplus, Haemaphysalis longicornis, Ixodes granulatus, Bacterial community diversity, High-throughput sequencing

中图分类号:

R384.41

管毓威, 向昱龙, 周敬祝, 罗小龙, 孔雪雪, 张燕, 胡勇, 梁文琴. 贵州省3个自治州寄生蜱细菌群落多样性研究[J]. 中国寄生虫学与寄生虫病杂志, 2025, 43(3): 370-376.

GUAN Yuwei, XIANG Yulong, ZHOU Jingzhu, LUO Xiaolong, KONG Xuexue, ZHANG Yan, HU Yong, LIANG Wenqin. Investigation of bacterial community diversity in parasitic ticks from three autonomous prefectures in Guizhou Province[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2025, 43(3): 370-376.

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样品组别 Sample group	香农指数 Shannon index	Chao指数 Chao index	Ace指数 Ace index	辛普森指数 Simpson index	均匀度指数 Evenness index	覆盖率/% Coverage/%
QDN-Rm	0.517	439.437	444.157	0.874	0.089	99.84
QDN-Hl	4.868	568.481	567.479	0.028	0.770	99.99
QN-Rm	0.633	202.604	205.282	0.737	0.128	99.91
QN-Hl	2.654	482.231	477.958	0.294	0.435	99.88
QXN-Rm	0.717	471.635	467.065	0.812	0.121	99.84
QXN-Ig	2.353	526.959	520.185	0.290	0.384	99.80

贵州省3个自治州寄生蜱细菌群落多样性研究

Investigation of bacterial community diversity in parasitic ticks from three autonomous prefectures in Guizhou Province

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