Background: Human subcutaneous myiasis is an endemic, understudied disease in Ethiopia. Objective: The aim of this study was to assess knowledge, perceptions and practices in different urban and rural communities of Jimma Zone, southwestern Ethiopia, related to human myiasis. Method: A cross-sectional, descriptive, comparative study was carried out during March -June 2005 in 5 urban and 3 rural kebeles in Jimma Zone. Kebeles were chosen by random sampling and a total of 1,272 households were selected by systematic random sampling technique for interview. Results: Most subjects (60.0%) mentioned lying on damp soil as the cause of myiasis. More urban than rural communities who wore un-ironed clothes had myiais infestation (p< 0.05). Myiasis was reported to be common in the lower extremities (65.6%), upper extremities (56.7%), abdomen (48.9%) and back (34.9%). A total of 86% dwellers responded that myiasis was more common during the wet season. More than half of the interviewees were knowledgeable about the clinical presentation of the disease. The most common signs and symptoms were itching (94.0%), pus discharge (82.9 %) and papules (79.5%). The majority (87.9%) of the study subjects treated infestations with hot matches. Wearing dry clothes (38.6%) was reported to be effective preventive measure. Nearly 64.8% of the rural and 68.7% of the urban communities knew that human subcutaneous myiasis is preventable. Conclusion: Due to the low levels of knowledge about myiasis, we recommend that community health education programs be developed through the health services to raise the awareness levels about this infestation to the general population.