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



Khoo, Jing Jing, 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) (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).

This is the latest version of this item.

[img] Text
microorganisms-08-00988.pdf - Published Version

Download (3MB) | Preview

Abstract

<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
Depositing User: Symplectic Admin
Date Deposited: 22 Jul 2020 08:45
Last Modified: 03 Mar 2021 10:15
DOI: 10.3390/microorganisms8070988
Open Access URL: https://doi.org/10.3390/microorganisms8070988
URI: https://livrepository.liverpool.ac.uk/id/eprint/3094788

Available Versions of this Item