Isolation and propagation of a Spiroplasma sp. from Slovakian Ixodes ricinus ticks in Ixodes spp. cell lines.
Isolation and propagation of a Spiroplasma sp. from Slovakian Ixodes ricinus ticks in Ixodes spp. cell lines.
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DOI：
10.1016/j.ttbdis.2015.05.002
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发表时间：
2015-07
影响因子：
3.2
通讯作者：
Kazimirova M
中科院分区：
文献类型：
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作者：
Bell-Sakyi L;Palomar AM;Kazimirova M
硬蜱属已知蜱偶尔携带螺原体-柔膜菌纲螺旋支原体;斯洛伐克先前的一项研究报告称，硬蜱螺原体的总患病率为3%。在本研究中，未进食的成年I。将从斯洛伐克西南部的植被中收集的蓖麻蜱加入一组来自I.蓖麻和肩突硬蜱胚胎。在随后的16-18个月内，通过制备和检查Giemsa染色的细胞离心涂片来监测培养物。2-3个月后，在两种蜱种属的培养物中检测到螺原体样微生物，并在适当细胞系的新鲜未感染细胞上传代培养7次。使用靶向16 S rRNA、ITS和rpoB基因片段的PCR测定进行的分子分析证实了微生物为螺原体属，与S.伊克索迪斯从两个不同场合收集的三个不同蜱池中分离的螺原体序列对于所有三种测试基因都是相同的。
Ixodes spp. ticks are known to occasionally harbour spiroplasmas – helical mycoplasmas in the class Mollicutes; a previous study in Slovakia reported an overall prevalence of Spiroplasma ixodetis of 3% in Ixodes ricinus. In the present study, extracts of unfed adult I. ricinus ticks collected from vegetation in south-western Slovakia were added to a panel of cell lines derived from I. ricinus and Ixodes scapularis embryos. The cultures were monitored by preparation and examination of Giemsa-stained cytocentrifuge smears at intervals over the subsequent 16–18 months. Spiroplasma-like microorganisms were detected in cultures of both tick species after 2–3 months and subcultured onto fresh, uninfected cells of the appropriate cell line up to seven times. Molecular analysis using PCR assays targeting fragments of the 16S rRNA, ITS and rpoB genes confirmed the identity of the microorganisms as a Spiroplasma sp., with between 98.9% and 99.5% similarity to S. ixodetis. The sequences of the spiroplasmas isolated from three different pools of ticks collected on two different occasions were identical for all three genes tested.