For the general population, food hygiene is a growing public health concern. Lack of awareness of food hygiene knowledge and practices will lead to poor food handling, which may result in early mortality, food-borne illnesses, and death. To evaluate food handlers’ knowledge, practices, and related aspects in various food enterprises in Nekemte town, Ethiopia, this study was designed. About 360 food handlers in the study area participated in a cross-sectional study that was based in an institution. Using methodical sampling approaches, the necessary sample was selected. Data were gathered using a semistructured questionnaire. For analysis, the data were entered in SPPSS version 20.0. According to the study results, 57.8% and 35.75% of food handlers in the study area had inadequate food hygiene knowledge and practices, respectively. Secondary school or above: 2.42 (95% CI: 1.13, 3.56), training: 4.65 (95% CI: 1.847, 11.74), experience of 1–5 years: 2.12 (95% CI: 1.283, 3.83), and experience of more than 5 years: 2.11 (95% CI: 1.183, 5.34) were variables that significantly predicted knowledge of food hygiene. Similarly, secondary education or higher: 2.19 (95% CI: 1.202, 4.83), experience of more than 5 years: 2.4 (95% CI: 1.12, 6.96), knowledge of food hygiene: 2.61 (95% CI: 2.14, 4.56), and training 3.3 (95% CI: 2.32, 4.76) were correlated with food hygiene practice of food handlers. To improve food hygiene knowledge and practice as well as to overcome their associated factors, such as educational status, training, work experience, and knowledge of food handlers, all relevant bodies should apply all feasible interventions through behavioral change communication.