Wijegunawardana, N.D.A.D.Silva Gunawardene, Y.I.N.Chandrasena, T.G.A.N.Dassanayake, R.S.Gunathilaka, P.A.D.H.N.Kittayapong, P.Xi, Z.Bourtzis, K.Abeyewickreme, W.2025-11-032025-11-032016-11-05Proceedings of the Peradeniya University International Research Sessions (iPURSE) – 2016, University of Peradeniya, P 303978-955-589-225-4https://ir.lib.pdn.ac.lk/handle/20.500.14444/5934During the last ten years, intensive research efforts have provided evidence that insect symbionts like Wolbachia can be useful tools for the control of major agricultural pests and disease vectors, including Aedes mosquitoes. The objective of the present study was to determine and characterize the presence of Wolbachia infections in wild mosquito populations in Sri Lanka in order to identify suitable strains which may be useful in the combined SIT / IIT approach. Adult mosquitoes were collected from 15 selected districts in Sri Lanka namely; Gampaha, Colombo, Galle, Matara, Hambanthota, Jaffna, Mannar, Ampara, Trincomalee, Batticaloa, Anuradhapura, Kandy, Kegalle, Badulla and Nuwara Eliya from October, 2014 to March, 2016. Mosquitoes were identified into the species leveland stored in -20 ⁰ C freezer after labeling until taken for molecular analysis. The field caught mosquito specimens were processed for the genomic DNA extraction individually using Qiagenkits (Qiagen DY14, Hilden, Germany). The DNA amplification was carried out using a specific pair of primersthat recognizes the 16S rRNA gene of Wolbachia. Primers which amplify the gene encoding for the major Wolbachia surface protein (wsp) were also used for screening in order to confirm the results. Two negative controls (Wolbachia un-infected mosquito DNA and milli-Q water) and one positive control (Wolbachiadouble infected Aedes albopictus Thailand strain) were used at each Polymerase Chain Reaction (PCR). Each PCR product of 5μl was subjected to gel electrophoresis and visualized in gel image system. The results were compared with the marker (100bp ladder) in an attempt to identify band sizes of about 1 kb and 540-632bp for the 16S rRNA and wspgenes, respectively. The experiment was conducted for 3, 24, 2, 5, 16, 7 and 21 species from each mosquito genus Tripteroides, Anopheles, Toxorhynchites, Mansonia, Aedes, Armigerus and Culex respectively. For confirmation of the results, each PCR reaction was repeated three times.PCR amplicons were further analyzed by nucleotide sequencing, Blastn search and sequence alignment. Accordingly, a total of 78 mosquito species were screened and twelve (12) of them namely; Mansoniaindiana, Mn. uniformis, Mn. annulifera, Aedesalbopictus, Ae. pseudalbopictus, Armigeressubalbatus, Ar. flavus, Culexgelides, Cx. quinquefaciatus, Cx.sinensisandCx.sitienswere positive by both PCR assays. None of the Anopheles species or Aedesaegypti was found infected with Wolbachia. Sequence analysis indicated significant genetic diversity among the various Wolbachiastrains detected.en-USWolbachiaMosquitoDNAPolymerase Chain Reaction (PCR)rRNAArticle