 The key to the effective management of healthcare wastes is segregation of the waste at the point of generation; no matter what final strategy for treatment and disposal of wastes is selected, it is critical that waste streams are separated. In Ethiopia, healthcare waste segregation practice among healthcare workers is overlooked and scarcely addressed in the scientific literature. This hospital-based cross-sectional study was, therefore, conducted to assess healthcare waste segregation practice and its correlate among healthcare workers in Bale zone, southeast Ethiopia.