Title : Comparison of leishmania culture in a new culture medium with NNN and RPMI1640 culture medium
Introduction: Leishmania is a common zoonotic parasite that cause leishmaniasis which has been recognized as a global issue in 98 countries. One of the most reliable methods to diagnose disease is parasite culturing, the most common media used are NNN and RPMI1640 but these media are expensive and difficult to prepare. In this study a new culture medium was compared with NNN and RPMI1640.
Methods: This study is a laboratory study. To prepare the new culture medium, we used agar, ground semolina, dextrose, blood, gentamicin, distilled water and 33.33% dextrose water serum. NNN medium was made based on standard protocol and RPMI1640 was purchased. After culturing of amastigote and promastigote, the number of parasites counted daily for 28 days and results was analyzed with Mann-Whitney test.
Result: Finally, in new culture medium Leishmania amastigotes didn’t transform to promastigotes. But, after inoculation of Leishmania promastigote, it was found the number of promastigote in the new culture medium was close to RPMI1640 until 8th day. But from the eighth day onwards, the number of parasites in the new culture medium decreased and on the sixteenth day it reached zero while the parasites continued to grow in RPMI1640. According to the result of the Mann-Whitney test, there was no significant difference between the two medium until the 8th day. From the 9th day onwards, there was a significant difference between the two medium (P > 0.05).
Conclusion: Eventually, in this study, it was found that the new culture medium is not suitable for the transformation of amastigotes into promastigotes, but it is suitable for the cultivation, growth and survival of promastigotes, like¬ as RPMI1640. Therefore, the new medium is benefitfull and cost effective in comparison to RPMI1640 for research on Leishmania promastigotes.