An analysis of bacterial community composition and diversity in Ixodes persulcatus in Inner Mongolia, China

doi:10.11853/j.issn.1003.8280.2019.06.003

Abstract

Abstract: Objective To investigate the bacterial community composition and population diversity in Ixodes persulcatus. Methods Roche 454 high-throughput sequencing technology was applied to determine the highly variable V3 segment of 16S rDNA gene of I. persulcatus in Inner Mongolia. Meanwhile, analyses were performed on bacterial community composition, population diversity, dominant bacterial population, and distribution of common pathogens. Results A total of 22 727 sequences and 1 570 operational taxonomic units were obtained for analysis. The coverage of all samples was greater than or equal to 0.95, which indicated that the number of sequences for sequencing was sufficient for covering basically all the bacterial species. Among the samples, male ticks had the highest richness index and diversity index (2 431 and 4.11, respectively), followed by female ticks and eggs. The dominant bacteria in I. persulcatus were Proteobacteria, Firmicutes, and Actinobacteria, sequentially, among which Alphaproteobacteria showed the highest relative abundance. On the genus level, the composition of genera with higher relative abundance varied greatly between different types of samples. Bacteria found in the samples were pathogens (i.e., Candidatus Rickettsia tarasevichiae, Spiroplasma, and Borrelia miyamotoi), environmental bacteria (i.e., Chryseobacterium, Erwinia, and Brevundimonas), and maternally inherited symbiont (i.e., Montezuma). Conclusion There are relatively significant differences in bacterial community composition and abundance between different I. persulcatus samples in Inner Mongolia.

Key words: Ixodes persulcatus, Bacterial community composition, Diversity, 16S rDNA, Roche 454 high-throughput sequencing

摘要： 目的了解全沟硬蜱细菌的群落结构及种群多样性。方法应用罗氏454高通量测序技术对内蒙古地区全沟硬蜱细菌16S rDNA基因的V3高变异区段进行测定，分析其细菌的群落结构、种群多样性、优势菌群以及常见病原体的分布。结果获得用于分析的序列和操作分类单元（OTUs）数目分别为22 727条和1 570个；所有样本的覆盖度均≥ 0.95，表明本次测序量充分，基本可覆盖所有细菌物种。其中，雄蜱样本的丰富度指数、多样性指数最高，分别为2 431和4.11；其次为雌蜱和卵。全沟硬蜱中优势菌群依次为变形菌门、厚壁菌门、放线菌门，其中以α-变形菌纲相对丰度最高；在属水平上，不同类别样本中相对丰度较高的菌属组成差异较大。在样本中检测到致病菌塔拉塞维奇立克次体、螺原体和宫本疏螺旋体，环境细菌如金黄杆菌属、欧文菌、短波单胞菌等，以及母系遗传共生菌（Montezuma）。结论内蒙古地区全沟硬蜱不同样本间细菌群落和丰度具有较大差异。

关键词: 全沟硬蜱, 细菌群落结构, 多样性, 16S rDNA, 罗氏454高通量测序

CLC Number:

R384.4
R37

LI Si-si, ZHANG Xiao-yu, ZHANG Yan-kai, LIU Jing-ze. An analysis of bacterial community composition and diversity in Ixodes persulcatus in Inner Mongolia, China[J]. Chines Journal of Vector Biology and Control, 2019, 30(6): 607-612.

李思思, 张晓雨, 张艳凯, 刘敬泽. 内蒙古地区全沟硬蜱细菌群落结构及多样性分析[J]. 中国媒介生物学及控制杂志, 2019, 30(6): 607-612.

References

[1] 张艳凯,刘敬泽. 蜱类病原体和共生菌多样性及其作用[J]. 应用昆虫学报,2019,56(1):1-11. DOI:10.7679/j.issn.2095-1353.2019.001.
[2] Eremeeva ME,Oliveira A,Moriarity J,et al. Detection and identification of bacterial agents in Ixodes persulcatus Schulze ticks from the north western region of Russia[J]. Vector Borne Zoonotic Dis,2007,7(3):426-436. DOI:10.1089/vbz.2007.0112.
[3] Kurilshikov A,Livanova NN,Fomenko NV,et al. Comparative metagenomic profiling of symbiotic bacterial communities associated with Ixodes persulcatus,Ixodes pavlovskyi and Dermacentor reticulatus ticks[J]. PLoS One,2015,10(7):e0131413. DOI:10.1371/journal.pone.0131413.
[4] Zhang XC,Yang ZN,Lu B,et al. The composition and transmission of microbiome in hard tick,Ixodes persulcatus,during blood meal[J]. Ticks Tick-Borne Dis,2014,5(6):864-870. DOI:10.1016/j.ttbdis.2014.07.009.
[5] 程成. 黑龙江口岸蜱媒病原的多样性及其复合感染调查[D]. 北京:中国人民解放军军事医学科学院,2017.
[6] 潘玉平,杨吉飞,牛庆丽,等. 黑龙江省全沟硬蜱携带伯氏疏螺旋体广义种和斑点热群立克次体的研究[J]. 中国兽医科学,2017,47(1):31-37. DOI:10.16656/j.issn.1673-4696.2017.01.005.
[7] Andreotti R,de León AAP,Dowd SE,et al. Assessment of bacterial diversity in the cattle tick Rhipicephalus (Boophilus) microplus through tag-encoded pyrosequencing[J]. BMC Microbiol,2011,11:6. DOI:10.1186/1471-2180-11-6.
[8] Narasimhan S,Fikrig E. Tick microbiome:the force within[J]. Trends Parasitol,2015,31(7):315-323. DOI:10.1016/j.pt.2015.03.010.
[9] Greay TL,Gofton AW,Paparini A,et al. Recent insights into the tick microbiome gained through next-generation sequencing[J]. Parasite Vector,2018,11:12. DOI:10.1186/s13071-017-2550-5.
[10] Bonnet SI,Binetruy F,Hernández-Jarguín AM,et al. The tick microbiome:why non-pathogenic microorganisms matter in tick biology and pathogen transmission[J]. Front Cell Infect Microbiol,2017,7:236. DOI:10.3389/fcimb.2017.00236.
[11] Yan P,Qiu ZX,Zhang TT,et al. Microbial diversity in the tick Argas japonicus (Acari:Argasidae) with a focus on Rickettsia pathogens[J]. Med Vet Entomol,2019,33(3):327-335. DOI:10.1111/mve.12373.
[12] Wang Q,Garrity GM,Tiedje JM,et al. Naïve Bayesian Classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy[J]. Appl Environ Microbiol,2007,73(16):5261-5267. DOI:10.1128/AEM.00062-07.
[13] 杨瑞馥,陶天申,方呈祥,等. 细菌名称双解及分类词典[M]. 北京:化学工业出版社,2011:10.
[14] Mirza OA. 利用反向线状印迹及高通量技术分别检测蜱体内梨形虫及全沟硬蜱体内细菌研究[D]. 北京:中国农业科学院,2016.
[15] 韩娜,张琳,张雯,等. 长角血蜱细菌群落结构及多样性研究[J]. 中国媒介生物学及控制杂志,2016,27(5):426-431. DOI:10.11853/j.issn.1003.8280.2016.05.002.
[16] 邹安迪. 褐黄血蜱的带菌分析[D]. 长沙:湖南农业大学,2016.
[17] Yu ZJ,Wang RR,Li NX,et al. Microbial diversity of the Tibetan tick Haemaphysalis tibetensis (Acari:Ixodidae)[J]. Exp Appl Acarol,2017,73(2):237-244. DOI:10.1007/s10493-017-0179-x.
[18] Maina AN,Jiang J,Omulo SA,et al. High prevalence of Rickettsia africae variants in Amblyomma variegatum ticks from domestic mammals in rural western Kenya:implications for human health[J]. Vector Borne Zoonot,2014,14(10):693-702. DOI:10.1089/vbz.2014.1578.
[19] Lopez-Velez R,Palomar AM,Oteo JA,et al. Novel Candidatus Rickettsia species detected in Nostril tick from human,Gabon,2014[J]. Emerg Infect Dis,2015,21(2):325-327. DOI:10.3201/eid2102.141048.
[20] Shpynov S,Fournier PE,Rudakov N,et al. ‘Candidatus Rickettsia tarasevichiae’ in Ixodes persulcatus ticks collected in Russia[J]. Ann N Y Acad Sci,2003,990(1):162-172. DOI:10.1111/j.1749-6632.2003.tb07358.x.
[21] Inokuma H,Ohashi M,Jilintai,et al. Prevalence of tick-borne Rickettsia and Ehrlichia in Ixodes persulcatus and Ixodes ovatus in Tokachi district,Eastern Hokkaido,Japan[J]. J Vet Med Sci,2007,69(6):661-664. DOI:10.1292/jvms.69.661.
[22] Katargina O,Geller J,Ivanova A,et al. Detection and identification of Rickettsia species in Ixodes tick populations from Estonia[J]. Ticks Tick-Borne Dis,2015,6(6):689-694. DOI:10.1016/j.ttbdis.2015.06.001.
[23] Jia N,Zheng YC,Jiang JF,et al. Human infection with Candidatus Rickettsia tarasevichiae[J]. N Engl J Med,2013,369(12):1178-1180. DOI:10.1056/NEJMc1303004.
[24] Fang LQ,Liu K,Li XL,et al. Emerging tick-borne infections in mainland China:an increasing public health threat[J]. Lancet Infect Dis,2015,15(12):1467-1479. DOI:10.1016/S1473-3099(15)00177-2.
[25] Ravyn MD,Korenberg EI,Oeding JA,et al. Monocytic Ehrlichia in Ixodes persulcatus ticks from Perm,Russia[J]. Lancet,1999,353(9154):722-723. DOI:10.1016/S0140-6736(98)05640-2.
[26] 曹务春,张泮河,张习坦,等. PCR检测蜱中查菲埃立克体DNA及其序列分析[J]. 寄生虫与医学昆虫学报,1999,6(1):58-63.
[27] 高东旗,曹务春,张习坦,等. 应用半巢式PCR检测我国北方一些蜱种中的查菲埃立克体[J]. 中国媒介生物学及控制杂志,2000,11(3):220-224. DOI:10.3969/j.issn.1003-4692.2000.03.018.
[28] 刘欢欢. 吉林省和黑龙江省蜱种分布及3种重要蜱传微生物分子流行病学调查[D]. 长春:吉林农业大学,2017.
[29] Yu PF,Niu QL,Liu ZJ,et al. Molecular epidemiological surveillance to assess emergence and re-emergence of tick-borne infections in tick samples from China evaluated by nested PCRs[J]. Acta Trop,2016,158:181-188. DOI:10.1016/j.actatropica. 2016.02.027.
[30] Takano A,Toyomane K,Konnai S,et al. Tick surveillance for relapsing Fever Spirochete Borrelia miyamotoi in Hokkaido,Japan[J]. PLoS One,2014,9(8):e104532. DOI:10.1371/journal. pone.0104532.