Molecular Characterization and Genetic Diversity of Wolbachia Strains in Culex quinquefasciatus, Culex pipiensand Drosophila melanogaster

Wolbachia are maternally inherited intracellular bacteria that occur naturally in a wide range of arthropods, where they manipulate host reproductive biology and influence pathogen survival and transmission of parasites. Identification of local Wolbachia strains is necessary to select the best strains for use in the control of vector-borne diseases. In this study we are presenting the first molecular characterization and assessment of the genetic diversity of Wolbachia in Culex quinquefasciatus , Culex pipiens and Drosophila melanogaster from Islamabad region of Pakistan. We identified species morphologically and then screened them for natural Wolbachia infection using PCR amplification of wsp , wspA , wspB , ftsZ , and 16S rRNA markers. Infection rates were 92% in Cx. quinquefasciatus , 67% in Cx. pipiens , and 85% in D. melanogaster . Co-infections with Wolbachia supergroups A and B were detected in all three species. Phylogenetic analyses of wsp sequences placed Culex strains within the cosmopolitan w Pip group, closely related to strains from diverse geographic regions, whereas D. melanogaster strains clustered with the well-characterized w Mel strain. This study provides the first molecular detection of Wolbachia in Cx. pipiens from Pakistan and the first evidence of A + B supergroup co-infections across multiple insect hosts in the country. Detection of these strains needs to be validated through further experiments to assess host fitness and suitability for field application.