Background and Aims: Akabane disease causes epidemics of abortions, stillbirths and congenital malformations manifested as arthrogryposis and hydranencephaly or microanencephaly in sheep, goats and cattle. Akabane virus replicates in arthropods and is transmitted by either mosquitoes or midges. Outbreaks of the Akabane virus have been reported from many countries in Southeast Asia, Middle East, Africa and Australia. The aim of this study was to evaluate the prevalence of Akabane virus infection and correlation of this infection with host and environmental determinants in sheep in Khouzestan province, the Southwest of Iran. Materials and Methods: In this study, serum samples of 360 sheep were randomly collected from 6 cities of Khouzestan province and were examined by ELISA assay. Results: Seroprevalence of Akabane virus infection was 39.72% (95% CI: 34.67-44.78%). Statistical analysis showed history of recently abortion and breed of sheep are significantly associated with infection (p<0.05) but sex and age of sheep are not significantly associated with infection (p>0.05). Conclusion: The results of the present study confirm that the Akabane virus infection exists in Khouzestan province, Iran. Considering the local weather conditions and the facility of vector-borne transmission, the health authorities should take measures to prevent and control the infection.