Department of Rickettsiology, Biomedical Research Center, Slovak Academy of Sciences, Dúbravská cesta 9, 845 05 Bratislava, Slovakia.
Laboratory of Molecular Structure Characterization, Institute of Microbiology, Czech Academy of Sciences, Vídeňská 1038, 142 20 Prague, Czech Republic.
Int J Environ Res Public Health. 2022 Apr 4;19(7):4321. doi: 10.3390/ijerph19074321.
Although the cat flea, , has been identified as the primary vector of , additional flea, tick, mite, and louse species have also been associated with this bacterium by molecular means; however, the role of these arthropods in the transmission of has not been clarified. Here, we succeeded in culture isolation of from a host-seeking castor bean tick, , the most common tick in Slovakia. The bacterial isolation was performed on XTC-2 cells at 28 °C using the shell-vial technique. An evaluation of the growth properties was performed for both the XTC-2 and Vero cell lines. We observed in the infected host cells microscopically by Gimenez staining and immunofluorescence assay. The isolate was purified by gradient ultracentrifugation and visualized by electron microscopy. Fragments of the genes A, A, B, A, B, 4, E, and were amplified and compared with the corresponding sequences of the type strain URRWXCal2 and other culture -isolated strains. We did not detect any nucleotide polymorphisms; however, plasmid pRFδ, characteristic of the standard strain, was absent in our isolate. Herein, we describe the first successful isolation and characterization of a tick-derived strain "Danube", obtained from an nymph.
尽管猫蚤已被确定为主要的传播媒介,但通过分子手段还发现了其他跳蚤、蜱、螨和虱种也与这种细菌有关;然而,这些节肢动物在传播中的作用尚未阐明。在这里,我们成功地从宿主寻找的蓖麻蜱中培养分离出,这是斯洛伐克最常见的蜱。细菌分离是在 28°C 的 XTC-2 细胞上通过壳瓶技术进行的。我们在 XTC-2 和 Vero 细胞系上评估了生长特性。我们通过 Gimenez 染色和免疫荧光检测在感染的宿主细胞中观察到。分离株通过梯度超速离心纯化,并通过电子显微镜观察。扩增了基因 A、A、B、A、B、4、E 和 的片段,并与标准株 URRWXCal2 及其他分离株的相应序列进行比较。我们没有发现任何核苷酸多态性;然而,标准株特有的质粒 pRFδ在我们的分离株中不存在。在此,我们描述了首次成功分离和表征来自幼虫的蜱衍生菌株“多瑙河”。