基于高通量测序技术分析浙江省余姚市蜱中携带细菌菌群多样性

High-throughput sequencing based analysis on diversity of pathogens carried by ticks in Yuyao, Zhejiang

目的了解浙江省余姚市蜱中携带的病原体构成。方法对采集的蜱进行形态学分类,再用高通量测序技术测定蜱所携带细菌16S rDNA基因的V3~V4高变异区段序列,进行生物学信息分析。结果共监测到蜱481只,均为硬蜱科,分别为血蜱属的长角血蜱、扇头蜱属的血红扇头蜱和硬蜱属的中华硬蜱,优势种群为长角血蜱,占94.18%。21组蜱的平均有效序列reads为55 753条,序列平均长度457.82 bp,(G+C)平均含量为52.07%。共产生2 104个分类操作单元,其稀释曲线趋于平缓,样本文库的覆盖率>99%。在蜱携带的微生物中共检出9个门77个属,居前3位的门依次是厚壁菌门、变形菌门、蓝藻门。其中厚壁菌门的丰度最高,为蜱的优势菌群。还发现了5种人兽共患病原菌,分别是葡萄球菌属、不动杆菌属、立克次体属、假单胞菌属、链球菌属,以葡萄球菌属为主,最高丰度达98.76%。结论通过高通量测序技术可以全面准确地分析蜱所携带的微生物群落信息,为蜱媒传染病、新发动物疫病的防控和综合监测技术平台的建立提供依据。

Objective To investigate the composition and diversity of tick-borne pathogens in Yuyao, Zhejiang. Methods The collected ticks were classified morphologically, and then the highly variable V3–V4 regions of bacterial 16 S rDNA gene carried by ticks were sequenced by high throughput sequencing, and their biological information were analyzed. Results A total of 481 ticks were detected, which were Haemaphysalis longicornis, Rhipicephalus sanguineus and Ixodes sinensis, respectively. The predominant population was H. longicornis, accounting for 94.18%. The average effective sequence reads of 21 groups of ticks was 55 753. The average length of the sequence was 457.82 bp, and the average content of G+C was 52.07%. A total of 2 104 operational taxonomic units(OTU) were generated. The dilution curve tended to be smooth, and the coverage of the sample library was more than 99%. The results of colony composition showed that a total of 9 phylum in 77 genus were detected, and the top three were Firmicutes,Proteobacteria and Cyanobacteria. Among them, the abundance of Firmicutes was highest, and it has become the predominant flora of ticks. Five kinds of zoonotic bacteria were also found in the microorganisms carried by ticks, which were Staphylococcus,Acinetobacter, Rickettsia, Pseudomonas and Streptococcus. Among them, Staphylococcus was the main species with the highest abundance(98.76%). Conclusion The microbial community information carried by ticks can be analyzed comprehensively and accurately by high-throughput sequencing, providing evidences for the prevention and control of tick-borne infectious diseases, new animal diseases and the establishment of comprehensive surveillance platform.