蜱是吸血外寄生虫,可作为传播各种病原体的载体。这项研究的目的是评估蜱传细菌,无论是否致病,使用16SrRNA元编码在韩国分布的蜱中,并通过PCR确认结果。2021年,使用标记方法从韩国的四个省收集了任务蜱。在从61个滴答池(包括372个滴答)中汇集DNA后,使用MiSeq平台扩增和测序细菌16SrRNAV3-V4高变区。立克次体,埃里希亚,通过常规PCR和分子分析证实了内共生体Wolbachia。总的来说,收集了6907个壁虱(534个池),并鉴定为属于五个物种(Haemphysalisspp。,H.Longicornis,H.flava,一、日本,和A.Testudinarium)。通过16SrRNA元编码,鉴定了240个扩增子序列变体。优势类群为立克次体和柯西氏菌。此外,病原菌,如立克次体和埃里希氏菌,鉴定了内共生细菌,例如Wolbachia和Spiroplasma。进行聚合酶链反应(PCR)以确认立克次体的存在,埃里希亚,巴尔通体,和Wolbachia在个体蜱中。总的来说,534个池中的352个(65.92%)对至少一种筛选的蜱传细菌呈阳性。立克次体是最普遍的(61.42%),其次是Wolbachia(5.05%)。在4.86%的检测样品中检测到埃里希菌,而没有检测到巴尔通体。在这项研究中,16SrRNA元编码揭示了立克次体的存在,Wolbachia,还有埃里希亚,按照丰富的顺序,同时显示没有巴尔通体。这些结果被证实表现出与常规PCR相同的趋势。因此,基于汇集的大规模筛查研究,正如在这项研究中所应用的那样,将用于检查存在于各种宿主和载体中的新型或稀有病原体。
Ticks are blood-sucking ectoparasites that act as vectors for transmission of various pathogens. The purpose of this study was to assess
tick-borne bacteria, whether pathogenic or not, in ticks distributed in Korea using 16S rRNA metabarcoding and to confirm the results by PCR. Questing ticks were collected from four provinces in Korea in 2021 using the flagging method. After pooling the DNAs from the 61
tick pools (including 372 ticks), the bacterial 16S rRNA V3-V4 hypervariable region was amplified and sequenced using the MiSeq platform. Rickettsia, Ehrlichia, and the endosymbiont Wolbachia were confirmed by conventional PCR and molecular analysis. In total, 6907 ticks (534 pools) were collected and identified as belonging to five species (Haemaphysalis spp., H. longicornis, H. flava, I. nipponensis, and A. testudinarium). Through 16S rRNA metabarcoding, 240 amplicon sequence variants were identified. The dominant taxa were Rickettsiella and Coxiella. Additionally, pathogenic bacteria such as Rickettsia and Ehrlichia, endosymbiotic bacteria such as Wolbachia and Spiroplasma were identified. Polymerase chain reaction (PCR) was performed to confirm the presence of Rickettsia, Ehrlichia, Bartonella, and Wolbachia in individual ticks. Overall, 352 (65.92%) of 534 pools tested positive for at least one of the screened
tick-borne bacteria. Rickettsia was the most prevalent (61.42%), followed by Wolbachia (5.05%). Ehrlichia was detected in 4.86% of tested samples, whereas Bartonella was not detected. In this study, 16S rRNA metabarcoding revealed the presence of Rickettsia, Wolbachia, and Ehrlichia, in that order of abundance, while showing absence of Bartonella. These results were confirmed to exhibit the same trend as that of the conventional PCR. Therefore, large-scale screening studies based on pooling, as applied in this study, will be useful for examining novel or rare pathogens present in various hosts and vectors.