Studies on the level of knowledge, attitude and handling practice towards food safety would help to determine the associated factors of knowledge, attitude and practice towards food safety; however, these studies did not explicitly address food safety concerns related to fruit and vegetables in Ethiopia. Men and women could have different levels of knowledge, attitude, and handling practice; however, these gendered effects were not addressed in previous studies. Using data collected in 2021 from 311 farm households in Ethiopia, this study aimed to analyze the level of knowledge, attitude, and handling practice of fruit and vegetable and associated factors among male and female food handlers. Data was analyzed using descriptive statistics, spearman's correlation, and econometric models (logit, multinomial logit and ordered probit). Education has a positive significant effect on knowledge, attitude and handling practices of fruit and vegetable handlers. However, the effect was higher in the females. No correlation could be obtained between knowledge and practices for the total respondents. However, knowledge had a positive but weak significant association with practices in the female group. We found a significant positive correlation between knowledge and attitude. These findings indicated that food safety knowledge of fruit and vegetable handlers will influence their attitude and a positive attitude would influence the practice in safe handling of fruit and vegetables. It therefore may require more targeted campaigns (i.e. from awareness creation to behavior change) to increase the ability of the community members to adopt best practices while reducing the barriers associated with consuming unhealthy diets.