Isolation of Candidatus Rickettsia vini from Belgian Ixodes arboricola ticks and propagation in tick cell lines.
Isolation of Candidatus Rickettsia vini from Belgian Ixodes arboricola ticks and propagation in tick cell lines.
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DOI：
10.1016/j.ttbdis.2020.101511
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发表时间：
2020-11
影响因子：
3.2
通讯作者：
Palomar AM
中科院分区：
文献类型：
--
作者：
Al-Khafaji AM;Bell-Sakyi L;Fracasso G;Luu L;Heylen D;Matthysen E;Oteo JA;Palomar AM
瓦氏念珠菌最初是在西班牙的软体硬蜱中发现的，随后在包括比利时在内的其他几个西方古北区国家报告了这一发现。最近，在哺乳动物(Vero)细胞培养中，从捷克共和国浸泡过的雄性侧柏细胞中分离到该细菌，但尚未见该细菌在壁虱细胞中繁殖的报道。在这里，我们报告了从扁虱细胞系中分离出三株钙。采自比利时大山雀(Parus Main)巢中的华山山雀R.vini。在无菌条件下解剖1只雄性和2只充血雌性硬蜱的内脏，加入微型立克次体细胞系BME/CTVM23的培养物中，在28 °C下培养，2~15周后在Giemsa染色的细胞离心片中首次发现类立克次体细菌。其中两个分离株生长迅速，在BME/CTVM23细胞中传代后2-4周内破坏了扁虱细胞，而第三个分离株生长较慢，仅需4−-5个月的传代培养。细菌16S rRNA和立克次体GLTA、sca4、ompB、ompA和17-kDa基因的PCR扩增结果表明，3株分离物均为Ca。与已发表的CA有100%同源性。来自其他地理位置的R.vini序列。透射电子显微镜显示BME/CTVM23细胞胞浆内有典型的单一立克次体细菌。加州大学。从雄性奥氏硬蜱中分离到的维氏丝虫菌株，命名为Boshek1，在一组蓖麻硬蜱、肩部硬蜱和微小硬蜱细胞系以及Vero细胞中生长的能力进行了测试。Boshek1菌株生长迅速，引起严重的细胞病变，在微优R.microplus品系BME26、I.ricinus品系IRE11和Vero细胞中，在I.ricinus品系IRE/CTVM19中生长较慢，可能在I.ricinus品系IRE/CTVM20中建立了低水平的侵染，并在12周的观察期内未能感染四个I.肩藻品系中的任何一个。本研究证实了简化的壁虱器官-细胞共培养技术在壁传立克次体分离中的适用性。使用BME/CTVM23细胞。
Candidatus Rickettsia vini was originally detected in Ixodes arboricola ticks from Spain, and subsequently reported from several other Western Palearctic countries including Belgium. Recently, the bacterium was isolated in mammalian (Vero) cell culture from macerated male I. arboricola from Czech Republic, but there have been no reports of propagation in tick cells. Here we report isolation in a tick cell line of three strains of Ca. R. vini from I. arboricola collected from nests of great tits (Parus major) in Belgium. Internal organs of one male and two engorged female ticks were dissected aseptically, added to cultures of the Rhipicephalus microplus cell line BME/CTVM23 and incubated at 28 °C. Rickettsia-like bacteria were first seen in Giemsa-stained cytocentrifuge smears between 2 and 15 weeks later. Two of the isolates grew rapidly, destroying the tick cells within 2–4 weeks of onward passage in BME/CTVM23 cells, while the third isolate grew much more slowly, only requiring subculture at 4−5-month intervals. PCR amplification of bacterial 16S rRNA and Rickettsia gltA, sca4, ompB, ompA and 17-kDa genes revealed that all three isolates were Ca. R. vini, with 100 % identity to each other and to published Ca. R. vini sequences from other geographical locations. Transmission electron microscopy revealed typical single Rickettsia bacteria in the cytoplasm of BME/CTVM23 cells. The Ca. R. vini strain isolated from the male I. arboricola tick, designated Boshoek1, was tested for ability to grow in a panel of Ixodes ricinus, Ixodes scapularis and R. microplus cell lines and in Vero cells. The Boshoek1 strain grew rapidly, causing severe cytopathic effect, in the R. microplus line BME26, the I. ricinus line IRE11 and Vero cells, more slowly in the I. ricinus line IRE/CTVM19, possibly established a low-level infection in the I. ricinus line IRE/CTVM20, and failed to infect cells of any of four I. scapularis lines over a 12-week observation period. This study confirmed the applicability of the simple tick organ-cell line co-cultivation technique for isolation of tick-borne Rickettsia spp. using BME/CTVM23 cells.
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影响因子：
9.4
作者：
Massung, Robert F.;Levin, Michael L.;Kurtti, Timothy J.
通讯作者：
Kurtti, Timothy J.
影响因子：
3.2
作者：
Bell-Sakyi L;Palomar AM;Kazimirova M
通讯作者：
Kazimirova M
影响因子：
2.4
作者：
Heylen, D. J. A.;Van Oosten, A. R.;Matthysen, E.
通讯作者：
Matthysen, E.
影响因子：
5.1
作者：
Heylen, Dieter;Tijsse, Ellen;Sprong, Hein
通讯作者：
Sprong, Hein
影响因子：
2.1
作者：
Keskin, Adem;Koprulu, Tugba Kul;Tekin, Saban
通讯作者：
Tekin, Saban