The study is the result of analyzing 16 895 blood smears of cattle collected at 180 sites in the provinces of Manica, Sofala, Zambézia and Tete in Mozambique. Of the blood smears 73.9 % were from Manica, 11.8 % from Tete, 8.5 % from Sofala and 5.8 % from Zambézia; 75.6 % of these were collected from smallholder cattle. Infections with trypanosomes were highest in smallholder cattle from Sofala Province with 36.8 % of the 872 blood smears examined positive for trypanosomes, and lowest in cattle of commercial farmers in Manica Province with only 6.2 % of 2 252 blood smears being positive. Trypanosoma congolense was the predominant species, followed by Trypanosoma vivax and Trypanosoma brucei sensu lato. Trypanosoma brucei, which also infects humans, was more frequent in the districts of Buzi, Mutarara and Morrumbala with 15.1 %, 10.5 % and 9.8 % of all examined cattle in 2005 being infected with it, respectively. The results show a significant increase in the infection rate with trypanosomes compared with results obtained in previous years by the Regional Veterinary Laboratory in Manica Province and by the Regional Tsetse and Trypanosomiasis Control Programme in Zambézia, Tete and Sofala provinces.