The study was conducted from October 2015 to November 2015 in Gambela and Abobo districts of Gambela Peoples Regional State, southwestern Ethiopia. It was designed to avail information on prevalence of bovine trypanosomosis and apparent densities of vectors of the trypanosomosis. A parasitological study using buffy coat technique was employed for the determination of prevalence of trypanosomosis while baited mono pyramidal traps were used for the vector survey. A total of 862 cattle randomly selected from the study population were examined for the parasitological study. The result of parasitological study revealed that the overall prevalence of trypanosomosis was found to be 16.59%, 95% CI=14.10-19.08 indicating trypanosomosis is a serious problem in the area. Three Trypanosoma species were identified during the study period: Trypanosoma brucei, T. vivax and T. congolense. Highest trypanosome prevalence (18.67%) was seen in animals with poor condition than that of those with medium (16.76%) and good (14.20%) body condition for the concerned blood parasite but no significant difference was observed (P>0.05). Higher infection rate was occurred in male (18.35%) than female (14.79%) cattle without significant difference (P>0.05). Prevalence was significantly higher in adult animals (17.95%) than younger ones (7.27%) (P>0.05). The study suggested that mean PCV values of parasitaemic (21.13%) cattle was significantly lower than aparasitaemic (22.26%) animals (P<0.05). During entomological survey, four species of tsetse fly: G. tachnoides, G. morsitsns submorsitans, G. pallidipes and G. fuscipes fuscipes and three genera of other biting flies: Tabanus, Haematopota and Stomoxys spp. were caught. The overall apparent density of tsetse flies was 0.75 fly/trap/day. Trypanosomosis and tsetse fly pose great threat to cattle residing in study areas. Thus, appropriate intervention measures need to be taken.