First report on the presence of natural Wolbachia population from major malarial vector mosquitoes Anopheles culicifacies s.l., and Anopheles stephensi from Tamil Nadu, India

Wolbachia is an alpha-proteobacteria present in several arthropods. The present study focussed on the identification of Wolbachia in Anopheles culicifacies s.l., and Anopheles stephensi, which are wild malarial vector mosquitoes in India. A total of four hundred and eighty seven adult female mosquito samples were collected using taxonomic keys and confirmed by molecular analysis where 48.25% was Anopheles culicifacies s.l., and 51.75% was Anopheles stephensi. The presence of Wolbachia was identified using 16S rRNA, wsp and FtsZ genes, where nested PCR of 16S rRNA alone was successful and sequenced. Only seven mosquitoes (1.4%) were positive for Wolbachia. In silico and restriction digestion of 16S rRNA gene product using RsaI enzyme showed that the identified Wolbachia belongs to supergroup B, which is further supported by phylogenetic analysis. Low prevalence rate of natural Wolbachia was observed in An. culicifacies s.l. (1.7%) and An. stephensi (1.2%). This is the first report on the presence of Wolbachia in Anopheles culicifacies s.l. and Anopheles stephensi.