Staphylococcus aureus is a leading source of foodborne poisoning in the human population also critically severe mastitis in livestock. A total of 671 serum samples were collected and tested through ELISA assays. The results revealed that the total prevalence of S. aureus infection in dairy cattle was 29.1% (95% CI: 25.6-32.7). In different farms, the prevalence ranged from 13.9% (95% CI: 8.8-20.5) to 36.6% (30.4-43.2). Similarly, the mean sero-prevalence was 17.0% (13.1-21.4) and 41.6% (36.3-47.2) in male and female animals, respectively. However, at different ages, the mean prevalence of Staphylococcus aureus was found to be 14.3% (7.4-24.1) to 38.5% (30.8-46.6). Although in different years, the sero-prevalence was 27.5% (22.2-33.4) in 2019, 19.6% (13.7-26.7) in 2020 and 36.5% (30.6-42.7) in 2021. According to conditional stepwise logistic regression analysis, different farms, gender, age and year were observed as the critical risk factors affecting the prevalence. Our results may contribute to development of prevention and control strategies against this zoonotic pathogen in farm animals in this region.