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

doi:10.12140/j.issn.1000-7423.2025.03.011

Abstract

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

CLC Number:

R384.41

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.

Figures/Tables 3

Fig. 1

Fig. 2

References 33

[1]	Moraga-Fernández A, Muñoz-Hernández C, Sánchez-Sánchez M, et al. Exploring the diversity of tick-borne pathogens: The case of bacteria (Anaplasma, Rickettsia, Coxiella and Borrelia) protozoa (Babesia and Theileria) and viruses (orthonairovirus, tick-borne encephalitis virus and louping ill virus) in the European Continent[J]. Vet Microbiol, 2023, 286: 109892.
[2]	邵中军. 我国重要蜱传疾病及传播媒介研究概述[J]. 中华卫生杀虫药械, 2021, 27(4): 293-299.
	Shao ZJ. Overview of serious tick-borne diseases and vector ticks in China[J]. Chin J Hyg Insectic Equip, 2021, 27(4): 293-299. (in Chinese)
[3]	向昱龙, 周敬祝, 刘英, 等. 贵州省部分地区蜱及其携带细菌调查[J]. 中国媒介生物学及控制杂志, 2022, 33(1): 148-152.
	Xiang YL, Zhou JZ, Liu Y, et al. An investigation of ticks and tick-borne bacteria in some areas of Guizhou Province, China[J]. Chin J Vector Biol Control, 2022, 33(1): 148-152. (in Chinese)
[4]	Beard D, Stannard HJ, Old JM. Parasites of wombats (Family Vombatidae), with a focus on ticks and tick-borne pathogens[J]. Parasitol Res, 2021, 120(2): 395-409.
[5]	Chauvin A, Moreau E, Bonnet S, et al. Babesia and its hosts: Adaptation to long-lasting interactions as a way to achieve efficient transmission[J]. Vet Res, 2009, 40(2): 37.
[6]	Pollet T, Sprong H, Lejal E, et al. The scale affects our view on the identification and distribution of microbial communities in ticks[J]. Parasit Vectors, 2020, 13(1): 36.
[7]	Greay TL, Gofton AW, Paparini A, et al. Recent insights into the tick microbiome gained through next-generation sequencing[J]. Parasit Vectors, 2018, 11(1): 12.
[8]	Duron O, Morel O, Noël V, et al. Tick-bacteria mutualism depends on B vitamin synthesis pathways[J]. Curr Biol, 2018, 28(12): 1896-1902.e5.
[9]	Narasimhan S, Rajeevan N, Liu L, et al. Gut microbiota of the tick vector Ixodes scapularis modulate colonization of the Lyme disease spirochete[J]. Cell Host Microbe, 2014, 15(1): 58-71.
[10]	Boularias G, Azzag N, Galon C, et al. High-throughput microfluidic real-time PCR for the detection of multiple microorganisms in ixodid cattle ticks in northeast Algeria[J]. Pathogens, 2021, 10(3): 362.
[11]	向昱龙, 周敬祝, 张燕, 等. 贵州省少数民族自治州微小扇头蜱的宏基因组分析[J]. 中国媒介生物学及控制杂志, 2023, 34(3): 319-325.
	Xiang YL, Zhou JZ, Zhang Y, et al. Metagenomic analysis of Rhipicephalus microplus from minority autonomous prefectures in Guizhou province, China[J]. Chin J Vector Biol Control, 2023, 34(3): 319-325. (in Chinese)
[12]	Jia N, Wang JF, Shi WQ, et al. Large-scale comparative analyses of tick genomes elucidate their genetic diversity and vector capacities[J]. Cell, 2020, 182(5): 1328-1340.
[13]	Lu M, Tian JH, Pan XL, et al. Identification of Rickettsia spp., Anaplasma spp., and an Ehrlichia canis-like agent in Rhipicephalus microplus from Southwest and South-Central China[J]. Ticks Tick Borne Dis, 2022, 13(2): 101884.
[14]	Gómez GF, Isaza JP, Segura JA, et al. Metatranscriptomic virome assessment of Rhipicephalus microplus from Colombia[J]. Ticks Tick Borne Dis, 2020, 11(5): 101426.
[15]	Makenov MT, Toure AH, Korneev MG, et al. Rhipicephalus microplus and its vector-borne haemoparasites in Guinea: Further species expansion in West Africa[J]. Parasitol Res, 2021, 120(5): 1563-1570.
[16]	Cardoso ADS, Santos EGG, Lima ADS, et al. Terpenes on Rhipicephalus (boophilus) microplus: Acaricidal activity and acetylcholinesterase inhibition[J]. Vet Parasitol, 2020, 280: 109090.
[17]	Maldonado-Ruiz LP, Neupane S, Park Y, et al. The bacterial community of the lone star tick (Amblyomma americanum)[J]. Parasit Vectors, 2021, 14(1): 49.
[18]	张钰, 张科, 刘佳伟, 等. 普氏野马分布区域亚洲璃眼蜱的宏基因组分析与病原体评估[J]. 中国寄生虫学与寄生虫病杂志, 2024, 42(4): 439-446, 453.
	Zhang Y, Zhang K, Liu JW, et al. Metagenomic analysis and potential assessment of Hyalomma asiaticum in the distribution area of Przewalski’s horses[J]. Chin J Parasitol Parasit Dis, 2024, 42(4): 439-446, 453. (in Chinese)
[19]	Jiao J, Lu ZY, Yu YH, et al. Identification of tick-borne pathogens by metagenomic next-generation sequencing in Dermacentor nuttalli and Ixodes persulcatus in Inner Mongolia, China[J]. Parasit Vectors, 2021, 14(1): 287.
[20]	Misra BR, Kumar N, Kant R, et al. Abundance of ticks (Acari : Ixodidae) and presence of Rickettsia and Anaplasma in ticks infesting domestic animals from Northern India[J]. J Med Entomol, 2021, 58(3): 1370-1375.
[21]	Xiang LL, Poźniak B, Cheng TY. Bacteriological analysis of saliva from partially or fully engorged female adult Rhipicephalus microplus by next-generation sequencing[J]. Antonie Van Leeuwenhoek, 2017, 110(1): 105-113.
[22]	管毓威, 罗小龙, 周敬祝, 等. 贵州省部分地区微小扇头蜱微生物群落多样性及抗生素抗性基因的宏基因组分析[J]. 中国媒介生物学及控制杂志, 2024, 35(4): 394-400, 439.
	Guan YW, Luo XL, Zhou JZ, et al. Metagenomic analysis of microbial community diversity and antibiotic resistance genes of Rhipicephalus microplus in some areas of Guizhou Province, China[J]. Chin J Vector Biol Control, 2024, 35(4): 394-400, 439. (in Chinese)
[23]	Xiang YL, Zhou JZ, Yu FX, et al. Characterization of bacterial communities in ticks parasitizing cattle in a touristic location in southwestern China[J]. Exp Appl Acarol, 2023, 90(1/2): 119-135.
[24]	Wang Q, Guo WB, Pan YS, et al. Detection of novel spotted fever group Rickettsiae (Rickettsiales : Rickettsiaceae) in ticks (Acari : Ixodidae) in Southwestern China[J]. J Med Entomol, 2021, 58(3): 1363-1369.
[25]	Kim JY, Yi MH, Mahdi AAS, et al. iSeq 100 for metagenomic pathogen screening in ticks[J]. Parasit Vectors, 2021, 14(1): 346.
[26]	陈秋, 何贤海, 孟娇, 等. 贵州省罗甸县山羊体表寄生蜱携带柯克斯体属细菌的基因特征分析[J]. 中国媒介生物学及控制杂志, 2024, 35(4): 417-421.
	Chen Q, He XH, Meng J, et al. Analysis of genetic characteristics of Coxiella carried by parasitic ticks on goat body surface in Luodian County, Guizhou Province, China[J]. Chin J Vector Biol Control, 2024, 35(4): 417-421. (in Chinese)
[27]	刘子维, 李华锋. 辽宁省营口市长角血蜱携带立克次体和埃立克体调查[J]. 中国媒介生物学及控制杂志, 2024, 35(6): 714-718.
	Liu ZW, Li HF. Investigation of Rickettsia and Ehrlichia harbored by Haemaphysalis longicornis in Yingkou City, Liaoning Province, China[J]. Chin J Vector Biol Control, 2024, 35(6): 714-718. (in Chinese)
[28]	Tufts DM, Sameroff S, Tagliafierro T, et al. A metagenomic examination of the pathobiome of the invasive tick species, Haemaphysalis longicornis, collected from a New York City borough, USA[J]. Ticks Tick Borne Dis, 2020, 11(6): 101516.
[29]	Guizzo MG, Parizi LF, Nunes RD, et al. A Coxiella mutualist symbiont is essential to the development of Rhipicephalus microplus[J]. Sci Rep, 2017, 7(1): 17554.
[30]	Qi Y, Ai LL, Zhu CQ, et al. Wild hedgehogs and their parasitic ticks coinfected with multiple tick-borne pathogens in Jiangsu Province, Eastern China[J]. Microbiol Spectr, 2022, 10(5): e0213822.
[31]	Li J, Kelly P, Guo WN, et al. Molecular detection of Rickettsia, Hepatozoon, Ehrlichia and SFTSV in goat ticks[J]. Vet Parasitol Reg Stud Reports, 2020, 20: 100407.
[32]	Zhang RL, Zhang Q, Yu GF, et al. Metagenomic deep sequencing obtains taxonomic and functional profiles of Haemaphysalis longicornis that vary in response to different developmental stages and sexes[J]. Exp Appl Acarol, 2021, 83(2): 285-300.
[33]	Takano A, Ando S, Kishimoto T, et al. Presence of a novel Ehrlichia sp. in Ixodes granulatus found in Okinawa, Japan[J]. Microbiol Immunol, 2009, 53(2): 101-106.

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