DNA barcode identification and molecular detection of bluetongue virus in Culicoides biting midges (Diptera: Ceratopogonidae) from western Thailand.

Abstract Biting midges of the genus Culicoides Latreille are biological vectors of bluetongue virus (BTV), a member of family Reoviridae, genus Orbivirus. About 30 species of Culicoides have been identified as competent BTV vectors worldwide. Even though high seroprevalence of BTV has been reported among livestock ruminants from western Thailand, the Culicoides species which contribute to BTV transmission remain unclear. In the present study, Culicoides were collected from eight sampling sites, located in two BTV prevalent provinces in western Thailand. Adult Culicoides were identified using wing morphology and cytochrome c oxidase subunit I (COI) mtDNA molecular marker. A total of 9,677 Culicoides specimens belonging to 7 subgenera, 3 species groups, and 23 species were identified. After comparing sequencing results with available data from GenBank, COI sequences of five species were reported for the first time from Thailand. The most abundant potential BTV vector species collected were C. peregrinus, followed by C. orientalis, C. imicola, C. oxystoma, and C. fulvus. Out of 72 Culicoides pools, 9 pools (4 from C. orientalis, 2 from C. imicola, 2 from C. oxystoma, and 1 from C. fulvus) were positive by BTV RT-PCR analyses. These results are new to Culicoides BTV vector knowledge in Thailand and will contribute to further BTV studies in this particular region.