Thirty-four Leishmania isolates obtained from Bolivian and Peruvian patients infected with mucocutaneous leishmaniasis were characterized by isoenzyme electrophoresis using 10 enzymatic markers; all belonged to the subspecies L.b. braziliensis. Three isolates showed marked variation compared with the reference strain with respect to 5 or 6 enzymes. These variant isolates originated from patients with forms of the disease which were unresponsive to treatment.