Visceral leishmaniasis (VL), caused by Leishmania infantum, is an endemic zoonosis in Iran. In recent years, Leishmania infection in cats has been reported in several countries where leishmaniasis is present. The aim of this study was to survey Leishmania infection in cats and to detect its causative agents in VL endemic areas in Iran. Forty stray cats were captured from two areas where VL is endemic, Fars and East Azerbaijan provinces. Infection with Leishmania was evaluated by parasitological and molecular methods. Leishmania parasites were detected in 4 out of 40 cats (10%). The parasite was isolated from the spleen of three and the liver of one cat and mass cultivated in the culture medium. Molecular and isoenzyme characterization revealed that the parasites isolated from the four cats were all L. infantum. Considering the high rate of feline Leishmania infection in this study, it may be suggested that cats have an epidemiological role in areas where VL is endemic in Iran.

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