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

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Khoo, Jing Jing ORCID: 0000-0003-1285-6052, Kurtti, Timothy J, Husin, Nurul Aini, Beliavskaia, Alexandra ORCID: 0000-0002-1194-5689, Lim, Fang Shiang, 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) , AbuBakar, Sazaly, Makepeace, Benjamin L ORCID: 0000-0002-6100-6727 and Bell-Sakyi, Lesley ORCID: 0000-0002-7305-0477 (2020) Isolation and Propagation of Laboratory Strains and a Novel Flea-Derived Field Strain of Wolbachia in Tick Cell Lines. Microorganisms, 8 (7). E988-.

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Abstract

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

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:	18 Jan 2023 23:40
DOI:	10.3390/microorganisms8070988
Open Access URL:	https://doi.org/10.3390/microorganisms8070988
Related URLs:	Author Publisher
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3094788

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• Isolation and Propagation of Laboratory Strains and a Novel Flea-Derived Field Strain of Wolbachia in Tick Cell Lines. (deposited 20 Jul 2020 07:42)
  • Isolation and Propagation of Laboratory Strains and a Novel Flea-Derived Field Strain of Wolbachia in Tick Cell Lines. (deposited 22 Jul 2020 08:45) [Currently Displayed]