Isolation in Natural Host Cell Lines of <i>Wolbachia</i> Strains <i>w</i>Pip from the Mosquito <i>Culex pipiens</i> and <i>w</i>Pap from the Sand Fly <i>Phlebotomus papatasi</i>
Lesley Bell-Sakyi,
Alexandra Beliavskaia,
Catherine S. Hartley,
Laura Jones,
Lisa Luu,
Lee R. Haines,
James G. C. Hamilton,
Alistair C. Darby,
Benjamin L. Makepeace
Affiliations
Lesley Bell-Sakyi
Department of Infection Biology and Microbiomes, Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool, Liverpool L3 5RF, UK
Alexandra Beliavskaia
Department of Infection Biology and Microbiomes, Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool, Liverpool L3 5RF, UK
Catherine S. Hartley
Department of Infection Biology and Microbiomes, Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool, Liverpool L3 5RF, UK
Laura Jones
The Pirbright Institute, Pirbright, Surrey GU24 0NF, UK
Lisa Luu
Department of Infection Biology and Microbiomes, Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool, Liverpool L3 5RF, UK
Lee R. Haines
Vector Biology Department, Liverpool School of Tropical Medicine, Liverpool L3 5QA, UK
James G. C. Hamilton
Division of Biomedical and Life Sciences, Faculty of Health and Medicine, Lancaster University, Lancashire LA1 4YG, UK
Alistair C. Darby
Department of Infection Biology and Microbiomes, Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool, Liverpool L3 5RF, UK
Benjamin L. Makepeace
Department of Infection Biology and Microbiomes, Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool, Liverpool L3 5RF, UK
Endosymbiotic intracellular bacteria of the genus Wolbachia are harboured by many species of invertebrates. They display a wide range of developmental, metabolic and nutritional interactions with their hosts and may impact the transmission of arboviruses and protozoan parasites. Wolbachia have occasionally been isolated during insect cell line generation. Here, we report the isolation of two strains of Wolbachia, wPip and wPap, during cell line generation from their respective hosts, the mosquito Culex pipiens and the sand fly Phlebotomus papatasi. wPip was pathogenic for both new C. pipiens cell lines, CPE/LULS50 and CLP/LULS56, requiring tetracycline treatment to rescue the lines. In contrast, wPap was tolerated by the P. papatasi cell line PPL/LULS49, although tetracycline treatment was applied to generate a Wolbachia-free subline. Both Wolbachia strains were infective for a panel of heterologous insect and tick cell lines, including two novel lines generated from the sand fly Lutzomyia longipalpis, LLE/LULS45 and LLL/LULS52. In all cases, wPip was more pathogenic for the host cells than wPap. These newly isolated Wolbachia strains, and the novel mosquito and sand fly cell lines reported here, will add to the resources available for research on host–endosymbiont relationships, as well as on C. pipiens, P. papatasi, L. longipalpis and the pathogens that they transmit.
