A cross-sectional study was conducted among 239 randomly selected meat handlers working in butcher shop in southern Ethiopia to assess factors associated with meat hygiene knowledge and practices. A binary logistic regression analysis with a 95% confidence interval (CI) and a p-value < 0.05 was used to identify factors that were significantly associated with good level of meat hygiene knowledge and practices. The findings revealed that 38.5% [95% CI: 32.2–44.8%] and 25.1% [95% CI: 19.7–30.5%] of meat handlers have good levels of meat hygiene knowledge and practices, respectively. Good level of meat hygiene knowledge was significantly (p < 0.05) associated with educational level, having meat hygiene training, and having regular supportive supervision by health workers, whereas good level of meat handling practice was significantly associated with work experience, educational level, have regular supportive supervision by health professionals, and having meat hygiene training. In conclusion, the majority of meat handlers have poor knowledge and practices regarding meat hygiene among meat handlers. Educational level, meat hygiene training, and supportive supervision by a health professionals were all independent predictors of meat hygiene knowledge and practice among meat handlers. As a result, health professionals should give regular training, butcher shop inspections, and supportive supervision for meat handlers in order to improve meat hygiene knowledge and practices among meat handlers.