Akabane virus (AKAV), which causes Akabane disease, is an arthropod-borne virus (arbovirus) transmitted by Culicoides biting midges and mosquitoes. AKAV is an important pathogen that causes abortion and congenital anomalies in ruminants. In this study, we determined the prevalence of AKAV infection and identified possible viral vectors in Turkey’s Eastern Mediterranean region. The presence and prevalence of AKAV infection were assessed using serological and virological methods. Serologically, the prevalence of AKAV antibodies in cattle, sheep and goats were 44.74% (400/894), 22.90% (60/262) and 14.52% (63/434), respectively, while the total prevalence was 32.89% (523/1590). AKAV-specific nucleic acid amplicons were obtained by real-time RT-PCR from 1.13% (9/799) and 1.74% (5/288) of the cattle and sheep tested, respectively. No goats were positive for AKAV RNA. Overall, AKAV-specific nucleic acid amplicons were detected in 0.87% (14/1604) of the sampled ruminants. In addition, specimens of the assumed vector, Culicoides, were caught using light traps and identified. Ten Culicoides species were detected in the area, of which Culicoides schultzei complex was the dominant species although 32 specimens could not be identified at the species level. These were defined as Culicoides spp. AKAV nucleic acid was detected in C. schultzei, Culicoides longipennis and Culicoides circumscriptus. Phylogenetic analysis indicated two different AKAV genogroups (genogroups Ib and genogroups II) while potential AKAV vectors in this region are C. schultzei complex, C. longipennis and C. circumscriptus.