Trypanosoma brucei colonizes the tsetse gut via an immature peritrophic matrix in the proventriculus

The peritrophic matrix of blood-feeding insects is a chitinous structure that forms a protective barrier against oral pathogens and abrasive particles 1 . Tsetse flies transmit Trypanosoma brucei , which is the parasite that causes human sleeping sickness and is also partially responsible for animal trypanosomiasis in Sub-Saharan Africa. For this parasite to establish an infection in flies, it must first colonize the area between the peritrophic matrix and gut epithelium called the ectoperitrophic space. Although unproven, it is generally accepted that trypanosomes reach the ectoperitrophic space by penetrating the peritrophic matrix in the anterior midgut 2 – 4 . Here, we revisited this event using fluorescence- and electron-microscopy methodologies. We show that trypanosomes penetrate the ectoperitrophic space in which the newly made peritrophic matrix is synthesized by the proventriculus. Our model describes how these proventriculus-colonizing parasites can either migrate to the ectoperitrophic space or become trapped within peritrophic matrix layers to form cyst-like bodies that are passively pushed along the gut as the matrix gets remodelled. Furthermore, early proventricular colonization seems to be promoted by factors in trypanosome-infected blood that cause higher salivary gland infections and potentially increase parasite transmission. This paper shows that the parasite Trypanosoma brucei is able to establish infection in flies by penetrating the protective peritrophic matrix in the fly midgut at the place of matrix secretion in the proventriculus. Such early proventricular colonization is potentiated by factors that are present in trypanosome-infected blood ingested by the flies.