The aim of this study is to investigate the comparison of pain, stress and physical activity levels of academic staff working at the university according to the ergonomic conditions in their working environments. For this study, the International Physical Activity Questionnaire - Short form (IPAQ), Scandinavian Musculoskeletal Questionnaire and Perceived Stress Scale (PSS) were administered to 85 volunteer university employees. According to ROSA evaluations, the average physical activity level of academic staff whose working environment was ergonomic was statistically lower than that of academic staff whose work environment was not ergonomic (p = 0.018). Additionally, mean perceived stress levels were similar in both groups (p > 0.05). According to the Scandinavian Musculoskeletal System Survey, academic staff experienced the most pain in the neck region in the last 12 months, followed by pain in the back and waist regions. Among these pains experienced in the last 12 months, it was determined that pain in the neck and waist areas prevented individuals from their activities. The findings of this study show that the ergonomic conditions of the working environments of individuals working at universities are related to their physical activity levels.