Isolation and Propagation of Laboratory Strains and a Novel Flea-Derived Field Strain of <i>Wolbachia</i> in Tick Cell Lines.

Khoo, Jing Jing ORCID: 0000-0003-1285-6052, Kurtti, Timothy J, Husin, Nurul Aini, Beliavskaia, Alexandra ORCID: 0000-0002-1194-5689, Lim, Fang Shiang ORCID: 0000-0002-2650-7518, Zulkifli, Mulya Mustika Sari, Al-Khafaji, Alaa M, Hartley, Catherine, Darby, Alistair C ORCID: 0000-0002-3786-6209, Hughes, Grant L ORCID: 0000-0002-7567-7185
et al (show 3 more authors) (2020) Isolation and Propagation of Laboratory Strains and a Novel Flea-Derived Field Strain of <i>Wolbachia</i> in Tick Cell Lines. Microorganisms, 8 (7). E988-.

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<i>Wolbachia</i> are intracellular endosymbionts of several invertebrate taxa, including insects and nematodes. Although <i>Wolbachia</i> DNA has been detected in ticks, its presence is generally associated with parasitism by insects. To determine whether or not <i>Wolbachia</i> can infect and grow in tick cells, cell lines from three tick species, <i>Ixodes scapularis</i>, <i>Ixodes ricinus</i> and <i>Rhipicephalus microplus</i>, were inoculated with <i>Wolbachia</i> strains <i>w</i>Stri and <i>w</i>AlbB isolated from mosquito cell lines. Homogenates prepared from fleas collected from cats in Malaysia were inoculated into an <i>I. scapularis</i> cell line. Bacterial growth and identity were monitored by microscopy and PCR amplification and sequencing of fragments of <i>Wolbachia</i> genes. The <i>w</i>Stri strain infected <i>Ixodes</i> spp. cells and was maintained through 29 passages. The <i>w</i>AlbB strain successfully infected <i>Ixodes</i> spp. and <i>R. microplus</i> cells and was maintained through 2-5 passages. A novel strain of <i>Wolbachia</i> belonging to the supergroup F, designated <i>w</i>CfeF, was isolated in <i>I. scapularis</i> cells from a pool of <i>Ctenocephalides</i> sp. cat fleas and maintained in vitro through two passages over nine months. This is the first confirmed isolation of a <i>Wolbachia</i> strain from a flea and the first isolation of any <i>Wolbachia</i> strain outside the "pandemic" A and B supergroups. The study demonstrates that tick cells can host multiple <i>Wolbachia</i> strains, and can be added to panels of insect cell lines to improve success rates in isolation of field strains of <i>Wolbachia</i>.

Item Type: Article
Uncontrolled Keywords: Ctenocephalides, Malaysia, Wolbachia, flea, in vitro culture, phylogeny, tick cell line
Depositing User: Symplectic Admin
Date Deposited: 22 Jul 2020 08:45
Last Modified: 08 Feb 2024 15:15
DOI: 10.3390/microorganisms8070988
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