Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

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

Jing Jing Khoo
,
Timothy Kurtti
,
Nurul Aini Husin
,
Alexandra Beliavskaia
,
Fang Shiang Lim
,
Mulya Mustika Sari Zulkifli
,
Alaa Al-Khafaji
,
Catherine Hartley
,
Alistair Darby
,
Grant Hughes
ORCID logo ,
Sazaly AbuBakar
,
Benjamin Makepeace
,
Lesley Bell-Sakyi
* ORCID logo
Version 1 : Received: 10 June 2020 / Approved: 12 June 2020 / Online: 12 June 2020 (04:51:00 CEST)

How to cite: Khoo, J.J.; Kurtti, T.; Husin, N.A.; Beliavskaia, A.; Lim, F.S.; Zulkifli, M.M.S.; Al-Khafaji, A.; Hartley, C.; Darby, A.; Hughes, G.; AbuBakar, S.; Makepeace, B.; Bell-Sakyi, L. Isolation and Propagation of Laboratory Strains and a Novel Flea-Derived Field Strain of Wolbachia in Tick Cell Lines. Preprints 2020, 2020060141. https://doi.org/10.20944/preprints202006.0141.v1 Khoo, J.J.; Kurtti, T.; Husin, N.A.; Beliavskaia, A.; Lim, F.S.; Zulkifli, M.M.S.; Al-Khafaji, A.; Hartley, C.; Darby, A.; Hughes, G.; AbuBakar, S.; Makepeace, B.; Bell-Sakyi, L. Isolation and Propagation of Laboratory Strains and a Novel Flea-Derived Field Strain of Wolbachia in Tick Cell Lines. Preprints 2020, 2020060141. https://doi.org/10.20944/preprints202006.0141.v1

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.

Keywords

Wolbachia; tick cell line; Ctenocephalides; flea; Malaysia; in vitro culture; phylogeny

Subject

Biology and Life Sciences, Immunology and Microbiology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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