No sleeping sickness or nagana cases have been reported in Botswana since 1985. In view of several confirmed clinical cases of nagana and reports of heavy bovine mortality, a parasitological survey was conducted to determine the prevalence of trypanosome infection in cattle in Maun and Shakawe areas of Ngamiland district. Wet blood films, buffy coat and Giemsa-stained thick and thin blood smears were used to detect trypanosomes in animals. Overall,trypanosome infection rate was 15.98%, with 5.94% and 27.29% in Maun and Shakawe respectively. The urgent need to combat trypanosomosis in Ngamiland, particularly in the Shakawe area, is highlighted, and a 3-phase integrated tsetse control strategy for this disease problem is discussed.