In Cameroon, >90% of cattle are considered exposed to African animal trypanosomiasis (AAT) infection, with the presence of tsetse rendering cattle husbandry as a very difficult proposition. A systematic review of data on AAT and tsetse from 1990 to 2021 was conducted to develop a national atlas. The review identified 74 relevant scientific documents, with three pathogenic Trypanosoma species (Trypanosoma vivax, T. congolense and T. brucei s.l.) most frequently identified as causing AAT. Trypanosoma grayi, T. theileri, T. simiae and the human African trypanosomiasis causative agent T. brucei gambiense were also identified in a wide range of hosts. The tsetse fly fauna of Cameroon comprises nine species, with Glossina palpalis palpalis and G. fuscipes fuscipes the most widely distributed following their identification in seven and five of the 10 regions, respectively. Two species, Glossina nigrofusca and G. pallicera pallicera appeared to be rare and were restricted to both forest and protected areas. The presence of AAT is associated with the presence of tsetse in the livestock–human–wildlife interface of Cameroon. AAT occurs beyond the tsetse belts of the country where mechanical vectors are abundant. This study provides AAT and tsetse maps to support ongoing interventions in Cameroon.