Project description:BackgroundCharacterization of the microbial diversity and symbiont dynamics of ticks may help to understand the development of ticks and reveal new strategies to control tick-transmitted pathogens, which has not yet been explored in the Tibetan tick Haemaphysalis tibetensis. This tick species is widely distributed in the Tibetan Plateau, and is recognized as one of the primary parasites affecting domestic and wild animals.MethodsIn the present study, the endosymbionts of H. tibetensis were characterized using diagnostic polymerase chain reaction (diagnostic PCR), and further evaluated for tissue distribution and population dynamics at each developmental stage of ticks and in tissues at different reproductive statuses by real-time quantitative polymerase chain reaction (RT-qPCR).ResultsTwo symbionts were found in H. tibetensis, and named as CLS-Ht (Coxiella-like symbiont in H. tibetensis) and RLS-Ht (Rickettsia-like symbiont in H. tibetensis). They showed 100% infection rate in both females and males of H. tibetensis. CLS-Ht and RLS-Ht can be observed within eggs, larvae, nymphs and adults, which indicates vertical transmission in H. tibetensis. CLS-Ht was specifically distributed in the female ovaries and Malpighian tubules, whereas RLS-Ht was detected within ovaries, Malpighian tubules, salivary glands and midguts of the ticks. Real-time qPCR suggested that adult ticks carried the largest amount of CLS-Ht and RLS-Ht with CLS-Ht having a significantly higher presence in females than in males (P < 0.05), whereas the presence of RLS-Ht showed no significant differences between sexes. In the ovaries, CLS-Ht distribution reached a peak at one day post-engorgement, and then gradually declined to a lower level, whereas no change was observed in RLS-Ht. In Malpighian tubules, the amount of both symbionts displayed an increasing trend with time post-engorgement. In midguts and salivary glands, the amount of RLS-Ht showed no significant differences.ConclusionTwo novel endosymbionts (CLS-Ht and RLS-Ht) were characterized in H. tibetensis both showing a high prevalence and stable vertical transmission. The described tissue distribution and population dynamics might imply the important functions of these symbionts during the development and reproduction of ticks.