Department of Entomology, Kansas State University, Manhattan, KS, USA.
Central European Institute of Technology, Center for Zoonoses, University of Veterinary and Pharmaceutical Sciences, Brno, Czech Republic.
Parasit Vectors. 2021 Jan 14;14(1):49. doi: 10.1186/s13071-020-04550-z.
The lone star tick (Amblyomma americanum), an important vector of a wide range of human and animal pathogens, is very common throughout the East and Midwest of the USA. Ticks are known to carry non-pathogenic bacteria that may play a role in their vector competence for pathogens. Several previous studies using the high throughput sequencing (HTS) technologies reported the commensal bacteria in a tick midgut as abundant and diverse. In contrast, in our preliminary survey of the field collected adult lone star ticks, we found the number of culturable/viable bacteria very low.
We aimed to analyze the bacterial community of A. americanum by a parallel culture-dependent and a culture-independent approach applied to individual ticks.
We analyzed 94 adult females collected in eastern Kansas and found that 60.8% of ticks had no culturable bacteria and the remaining ticks carried only 67.7 ± 42.8 colony-forming units (CFUs)/tick representing 26 genera. HTS of the 16S rRNA gene resulted in a total of 32 operational taxonomic units (OTUs) with the dominant endosymbiotic genera Coxiella and Rickettsia (> 95%). Remaining OTUs with very low abundance were typical soil bacterial taxa indicating their environmental origin.
No correlation was found between the CFU abundance and the relative abundance from the culture-independent approach. This suggests that many culturable taxa detected by HTS but not by culture-dependent method were not viable or were not in their culturable state. Overall, our HTS results show that the midgut bacterial community of A. americanum is very poor without a core microbiome and the majority of bacteria are endosymbiotic.
孤星蜱(Amblyomma americanum)是一种重要的媒介,能够传播多种人类和动物病原体,在美国东部和中西部非常常见。蜱虫携带非致病性细菌,这些细菌可能在其作为病原体媒介的能力中发挥作用。先前使用高通量测序(HTS)技术的几项研究报告了蜱虫中肠共生菌的丰富度和多样性。相比之下,在我们对野外采集的成年孤星蜱的初步调查中,我们发现可培养/有活力的细菌数量非常低。
我们旨在通过平行的依赖培养和非依赖培养方法来分析 A. americanum 的细菌群落,并应用于单个蜱虫。
我们分析了在堪萨斯州东部采集的 94 只成年雌性蜱虫,发现 60.8%的蜱虫没有可培养的细菌,其余蜱虫仅携带 67.7±42.8 个菌落形成单位(CFU)/蜱虫,代表 26 个属。16S rRNA 基因的 HTS 总共产生了 32 个操作分类单元(OTU),其中优势内共生菌属为柯克斯体和立克次体(>95%)。其余 OTU 的丰度非常低,属于典型的土壤细菌类群,表明其来源于环境。
CFU 丰度与非依赖培养方法的相对丰度之间没有相关性。这表明,HTS 检测到的许多可培养类群,但未通过依赖培养方法检测到的类群,可能是不可培养的或处于不可培养状态。总体而言,我们的 HTS 结果表明,A. americanum 的中肠细菌群落非常贫瘠,没有核心微生物组,大多数细菌是内共生的。