SUMMARYDrosophila melanogasteris an important model system of immunity and parasite resistance, yet most studies use parasites that do not naturally infect this organism. We have studied trypanosomatids in natural populations to assess the prevalence and diversity of these gut parasites. We collected several species ofDrosophilafrom Europe and surveyed them for trypanosomatids using conserved primers for two genes. We have used the conserved GAPDH sequence to construct a phylogenetic tree and the highly variable spliced leader RNA to assay genetic diversity. All 5 of the species that we examined were infected, and the average prevalence ranged from 1 to 6%. There are several different groups of trypanosomatids, related to other monoxenous Trypanosomatidae. These may represent new trypanosomatid species and were found in different species of EuropeanDrosophilafrom different geographical locations. The detection of a little studied natural pathogen inD. melanogasterand related species provides new opportunities for research into both theDrosophilaimmune response and the evolution of hosts and parasites.</jats:p