Spiroplasma are vertically-transmitted endosymbionts of ticks and other arthropods. Field-collected Ixodes persulcatus have been reported to harbour Spiroplasma, but nothing is known about their persistence during laboratory colonisation of this tick species. We successfully isolated Spiroplasma from internal organs of 6/10 unfed adult ticks, belonging to the third generation of an I. persulcatus laboratory colony, into tick cell culture. We screened a further 51 adult male and female ticks from the same colony for presence of Spiroplasma by genus-specific PCR amplification of fragments of the 16S rRNA and rpoB genes; 100% of these ticks were infected and the 16S rRNA sequence showed 99.8% similarity to that of a previously-published Spiroplasma isolated from field-collected I. persulcatus. Our study shows that Spiroplasma endosymbionts persist at high prevalence in colonised I. persulcatus through at least three generations, and confirms the usefulness of tick cell lines for isolation and cultivation of this bacterium.

Centre for Infectious Animal Diseases Faculty of Tropical AgriSciences Czech University of Life Sciences Prague Czechia

Centre of Rickettsiosis and Arthropod Borne Diseases Hospital Universitario San Pedro CIBIR Logroño Spain

Department of Infection Biology and Microbiomes Institute of Infection Veterinary and Ecological Sciences University of Liverpool Liverpool United Kingdom

Institute of Parasitology Biology Centre of the Czech Academy of Sciences Ceske Budejovice Czechia

Science Center for Family Health and Human Reproduction Problems Irkutsk Russia

Veterinary Research Institute Brno Czechia

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