Cell cloning techniques and schizodeme analysis were used to detect mixtures of subpopulations in Leishmania parasites, isolated from humans and a reservoir host. Clones were obtained by plating promastigotes, at limited dilution, on solid medium. The resultant colonies were analysed by the restriction profile of kinetoplast DNA minicircles (schizodeme analysis). The efficiency of plating was around 40-60%, and subpopulations of different schizodemes, in stocks of Leishmania isolated from 2 cases of human cutaneous leishmaniasis were detected. The presence of different schizodemes in one isolate from a sylvatic animal suggested the possibility of a mixed natural infection.