Objective: The aim of this study is to determine the best liquid culture medium, which could reproduce highest number of Leishmania promastigotes to be potentially used in molecular studies, isoenzyme analyses and drug studies in the shortest time.

Method: Leishmania spp. strains stored in liquid nitrogen tanks were revitalized and inoculated into Novy-McNeal-Nicolle (NNN) culture medium. The isolates reproduced in the culture medium were genotyped via real time PCR, using primers and probes spesific to internal transcribed spacer-1 sequence. The isolates, obtained from the patients with cutaneous leishmaniasis in Turkey and were initially genotyped as Leishmania tropica, Leishmania major and Leishmania infantum/donovani, were inoculated into RPMI 1640, M199, Schneiders Medium, Nutrient Broth and Brain Heart (infusion) Broth culture media. Inoculated culture media were checked every day up to 12th day after inoculation and their reproduction density was assessed on hemocytometer.

Results: It has been observed that Leishmania tropica, L. major and L. infantum/donovani isolates, obtained from the patients with cutaneous leishmaniasis in Turkey, can be reproduced in RPMI 1640 culture medium better than M199, Schneider?s Medium, Nutrient Broth, Brain Heart Broth culture media, with much more promastigote reproduction. For all three Leishmania spp. inoculated promastigots reached the required reproduction density suitable for use beginning from the 8th day (p<0.05).

Conclusion: It has been concluded that RPMI 1640 was the most suitable medium when higher number of Leishmania promastigotes are needed, followed by preference for M199, Schneider?s Medium, Brain Heart Broth and Nutrient Broth media in that order.