The isoenzymatic typing of 16 stocks of Leishmania, isolated from Tunisian visceral leishmaniasis (VL) cases, revealed that all strains belonged to Leishmania (L.) infantum species. Although zymodeme MON-1 was predictably the most frequent (9 cases), it came as a surprise that L. infantum MON-24 was responsible of about third of cases. This latter zymodeme, while previously reported in Tunisia, Algeria and Spain, was assumed to be a dermotropic and the few cases of VL that it caused occurred always in HIV infected patients. L. infantum MON-80, occasionally reported during both cutaneous and VL of immunocompetent infants was identified in 2 patients. This report confirms that in addition to the more common L. infantum MON-1, zymodeme MON-24 has a substantial role in generating VL in immunocompetent infants in Tunisia.