A cross-sectional study to determine the prevalence of bovine trypanosomosis was carried out in Chena district, Kefa zone, southwest Ethiopia from September 2010 to January 2011. Blood samples were collected from 391 randomly selected local (zebu) breed cattle in three representative peasant associations (PAs). The buffy coat and Giemsa stained thin blood films examination techniques were used for parasite detection and identification. The packed cell volume (PCV) estimation was also conducted. From a total of 391 cattle, 6.9% (n = 27) of the animals were found positive for trypanosome infection. The trypanosome species observed across the study animals were T. congolense (4.89%), T. vivax (1.54%), and T. b. brucei (0.51%) as single infections. The infection rate of T. congolense and T. b. brucei varied significantly (P < 0.05). The statistical analysis revealed that no significant difference (P > 0.05) in infection rate was found between male (7.79%) and female (5.62%) animals. The prevalence was 2.24% and 8.29% in young and adult cattle, respectively with significant difference (P < 0.05) between them. The prevalence rate in good, medium and poor body conditioned animals were 7.28%, 0.78% and 13.39%, respectively with a statistical significant difference (P < 0.05) among them. The mean PCV of the infected animals (17.56%) appeared significantly (P < 0.05) lower than the non-infected (25.4%). Even if, the present study indicated a low prevalence of bovine trypanosomosis in the study areas, its impact on production and potential transmission to others should not be neglected.