Evaluation of hydatid cyst fluid as a substitute for fetal bovine serum (FBS) in culture of Leishmania major

Background: Isolation and characterization of Leishmania parasites is a necessary strategy for
control of Leishmaniasis. Free-cell culture media, rich with biologic fluids such as Fetal Bovine
Serum (FBS) are among the best media for culturing of the parasite. In our country, FBS is very
expensive and is not available everywhere. In this study, we evaluated Ovine Hydatid Cyst Fluid
(HCF) as a substitute for FBS in liquid culture media of Leishmania major.
Materials & Methods: Leishmania parasites were isolated from Balb/C mouse ulcer and
characterized by PCR. Of six tubes by triplicate (totally 18 tubes) add to each tube 800,000
promastigotes of Leishmania major and then add 1 cc of media including, once BHI (Brain Heart
Infusion broth), BHI plus 5% FBS, BHI plus 10% FBS, once Hydatid Cyst Fluid (HCF) , BHI plus
5% HCF and BHI plus 10%HCF , respectively. After 24h, 72h, 144h, 192 hours the number of
parasites in each tube counted and the means of them compared together.
Results: The result of this study showed that BHI plus HCF 10% medium could be use as a
suitable substitute till 72 hours for Fetal Bovine Serum (FBS) in liquid Culture of Leishmania major
parasites.
Conclusion: In this study, we have introduced the Ovine Hydatid Cyst Fluid (HCF) as
replacement for FBS in liquid Culture media.