Evaluation of three new culture media for the cultivation and isolation of Leishmania parasites.

The aim of this study was to establish novel culture media for Leishmania parasites with a potential of obtaining high amounts of promastigotes with long-term viability, and consisting of ingredients that were available in microbiology or parasitology laboratories. Other features of these media included no requirement for blood, FCS (Fetal calf serum) or erythrocyte lysate, inexpensiveness and easiness in preparation. In addition, aspiration samples obtained from cutaneous leishmaniasis (CL) suspected patients were cultivated in these media. Three culture media were prepared; trypticase beef extract hemoglobine (TBH) medium, including trypticase, beef extract and yeast extract as the protein source, glucose as the carbohydrate source, FeNH4 and bovine hemoglobine; Peptone-Yeast extract medium (PY), found to be effective in our previous studies for cultuvation of on Leishmania parasites, with bovine hemoglobine (PYH) and Brain Heart medium, containing bovine hemoglobine (BKH). The number of promastigotes were the highest on day 8 and 13 in RPMI 1640 and BKH medium, respectively. In TBH and PYH, the peak level of reproduction was between day 16 and 19, and it was found to be higher in TBH medium after the day 20. The number of promastigotes were found to be close in BKH, TBH and RPMI-1640 media and lower in PYH medium. Examination of the cultivation of the aqueous lesion specimens of the 10 CL-suspected cases in media revealed reproduction in 9 flasks of RPMI-1640 containing 10% FCS, 7 TBH, 6 BKH and 4 PYH. The differences between the culture media were not found to be statistically significant. These results suggested that, three liquid culture media, assessed in this study, with no requirement for FCS or erythrocyte lysate, were effective in the reproduction of promastigotes, and could be used effectively in the patient isolation and field studies, as well. Copyright 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim