BackgroundThe utilization of veterinary vaccines plays a vital role in preventing and managing animal diseases. However, the success of vaccination programmes relies on livestock farmers’ understanding, perspectives and behaviours regarding their application. Therefore, this study aims to evaluate the knowledge, attitudes and practices of livestock farmers in Southwest Ethiopia regarding the use of veterinary vaccines.MethodsThe study followed a cross‐sectional design conducted between October 2021 and October 2022. A sample of 476 livestock‐owning farmers, including those who raise dairy cattle, beef cattle and poultry, were interviewed from the districts of Shei Bench, Semen Bench, Sheko and Yeki.ResultsThe findings revealed that participants from Sheko, who had a poultry farm, were college graduates and resided in an urban area, were more likely to have good knowledge of veterinary vaccines. Similarly, participants who were male, from Semen Bench, had a poultry farm and had a high school education, exhibited a positive attitude towards veterinary vaccines. Furthermore, being male, from Semen Bench, having a poultry farm, and having a high school education were associated with increased veterinary vaccine use.ConclusionThis study highlights the importance of addressing gender differences and tailoring interventions based on geography and farming conditions in the area. These measures are crucial to improve practices related to veterinary vaccines for enhanced animal health and productivity. Prioritizing education, veterinary services and information dissemination is vital. However, further research and targeted interventions are needed to better comprehend underlying factors and implement effective strategies for diverse communities in different areas.