Isolation of Trypanosoma spp. from wild tsetse flies through procyclic expansion in Glossina morsitans centralis

Abstract Procyclic trypanosomes from wild tsetse flies were membrane-fed to Glossina morsitans centralis in order to develop an optimal technique for propagating field isolates. A 70% success rate was achieved in isolating Trypanosoma simiae and a variety of genotypes of T. congolense originating from G. pallidipes, G. brevipalpis and G. swynnertoni . Parasites matured into forms infective for mammals, and could be maintained by passage of gut forms to new groups of flies. In experiments with laboratory stocks, we also passaged immature gut infections of T. congolense and T. brucei from various tsetse species to G. m. centralis . The optimal technique was investigated for procyclic T. congolense through addition of various compounds to goat blood using G. m. centralis and G. m. morsitans as recipients. From these experiments, many approaches to procyclic expansion appeared possible. However, a simple and practical method based on the use of fresh goat blood for rapid feeding of G. m. centralis is recommended. Application of this technique should aid in the resolution of questions relating to the cryptic diversity of Nannomonas trypanosomes in diverse host and vector communities.