ABSTRACT Background Snakebites cause significant morbidity and mortality in Uganda. Effective management of snakebites requires knowledge of the appropriate first aid measures, as well as knowledge of the appropriate antivenom to use, but little is known about familiarity with effective snakebite management techniques and associated factors among healthcare practitioners (HCPs) in Uganda. Methods In May 2022, we collected data on sociodemographic characteristics, knowledge of snakebite first aid, envenomation signs, diagnosis and antivenom administration among 311 HCPs from two snakebite high-incidence districts in Uganda using a semi-structured questionnaire. Results Of the 311 HCPs, 64.3% had ever treated snakebite cases, 87.1% were confident to provide supportive treatment, but only 9.6% had ever been trained on snakebite management. Overall, 22.8% of HCPs had high knowledge of snakebite management. Higher education (at least degree vs certificate; PR=2.21 95% CI 1.508 to 4.56), older age (30–45 vs <30 y; PR=1.97, 95% CI 1.22 to 3.21) and previous training (PR=1.82, 95% CI 1.08 to 3.05) were associated with high knowledge of snakebite diagnosis and management. Conclusions Overall, knowledge of snakebite management was limited. Training, level of education and age of the HCP all had an impact on knowledge. Deliberate efforts are required to increase HCPs' knowledge of snakebite case care in high-burden regions to manage incident cases.