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A technique for culturing two species of trypanosomatid flagellates on blood-agar plates is described. Herpetomonas megaseliae and Crithidia harmosa were grown as discrete clone colonies on plates prepared in sterile petri dishes with a medium derived from that of Tobie et al. (1950, J. Parasitol. 36: 48-54). Plates were inoculated with 0.01 ml flagellate suspension and the inoculum was spread evenly over the surface of the agar with a sterile glass spreader. Plates were incubated in a high humidity chamber at 25 C and colonies of both species, easily visible to the unaided eye, appeared within 1 week. There was a virtual 1:1 ratio between calculated number of CFU (colony forming units) and actual CFU observed on the plates. Colony morphology differed between the two species. Herpetomonas megaseliae produced a translucent flattened colony with a raised center and raised spiral arms, while C. harmosa produced an opaque hemispherical colony. Applications of the culture technique include potential diagnosis of trypanosomatid infections and potential studies in trypanosomatid genetics.