Meat contamination during slaughter and sale affects 600 million people globally annually, leading to diseases and 420,000 deaths. However, evidence on hygienic practices in low-income countries, particularly in Ethiopia, limits the extent of this problem, highlighting the need for research on hygienic practices. The aim of this study was to assess the magnitude of meat hygiene practices and factors associated with poor practices among food handlers in Gambela town, Ethiopia. The study was an institution-based, cross-sectional study among food handlers. A total of 422 participants were chosen at random using systematic sampling. Data were collected using structured questions through interviews and observations. The study analyzed the prevalence of good hygiene practices and factors associated with meat hygiene practices using binary logistic regression. Statistical significance was determined using adjusted odds ratios, 95% confidence interval, and p-value of .05, and the model was checked using Hosmer and Lemeshow tests. The study result shows that good hygiene practices were only 40.6% (36.0–45.6) among meat handlers at butcher houses and restaurants in Gambela town. Low hygiene practices were significantly associated with the education level of food handlers (AOR: 2.5, 95%CI [1.17–5.41]) and meat hygiene training (AOR: 5.1, 95%CI [2.55–10.23]). Therefore, intensive food safety training for the workers, engaging educated professionals in food safety services, and mitigating the health risks and consequences of contamination demand intervention in the setting.