Potential roles of Culicoides spp. ( Culicoides imicola, Culicoides oxystoma ) as biological vectors of bluetongue virus in Yuanyang of Yunnan, P. R. China

plays a crucial role as an insect vector in the field of veterinary medicine. The transmission of significant viruses such as bluetongue virus (BTV) and African horse sickness virus (AHSV) by this insect poses a substantial threat, leading to the development of severe diseases in domestic animals. This study aimed to explore the species, identify their blood-meal sources, and assess the presence of BTV and AHSV carried by in Yuanyang County, Yunnan Province. The aim was to gain insights into the potential vectors of these two viruses and elucidate their potential roles in the transmission of pathogens. The midges were collected from cattle ( ), pig ( ), and goat ( ) pens in Yuanyang County, Yunnan Province in June 2020. Initial identification of midges was conducted through morphological characteristics, followed by molecular identification using the cytochrome C oxidase subunit I (COI) gene. The determination of blood-meal sources was accomplished using specific primers targeting the cytochrome (Cyt ) gene from potential hosts. BTV and AHSV RNA were identified in pools through the application of reverse transcriptase PCR and quantitative real-time PCR. Nucleotide homology and phylogenetic analysis were performed using MegAlign (DNAStar) and Mega 6.0 software. A total of 6,300 , consisting of , and , were collected from cattle, pigs, and goat pens. The engorgement rates for these species were 30.2%, 54.6%, 75%, and 66.7%, respectively. In the cattle pen, the prevailing species is (100%). In the pig pen, dominates (70%), with following at 30%. In the goat pen, holds the majority (45.45%), trailed by (25%), (20.45%), and (9.09%). These species were identified as feeding on cattle, pigs, goats, chickens ( ), and humans ( ). The positivity rates for BTV were 20.00% and 11.54% in blood-fed specimens of and , respectively. Conversely, the positivity rates for BTV in non-blood-fed specimens were 0.00% and 6.67% for and , respectively. BTV was not detected in and . The specimens (YY86) from that tested positive for BTV had the closest genetic relationship to YTS-4 isolated from Mangshi, Yunnan Province in 1996. All test results for the nucleic acid of AHSV were negative. The study reveals variations in the species distribution, community composition, blood sucking rate, and blood-feeding sources of across different habitats. Notably, and emerge as potential vectors for the transmission of BTV in local animals. Accordingly, this investigation provides crucial insig