Abstract
Leishmania parasite isolation from the human aspirates is always challenging due to most probability of the fungal contamination and the use of antifungal drug which could support the selective growth of the Leishmania parasite. In this study, we examine the effect of antifungal drug caspofungin on the promastigote stage of Leishmania donovani. Promastigote parasite was cultivated in M199 + 20% heat-inactivated fetal calf serum and plated in 96-well plates. Seven different concentrations of caspofungin (512 µg/ml to 8 µg/ml) were exposed to parasites, and 50% inhibitory concentration (IC50) was calculated. Candida spp. was used in the experiments to know the efficacy of caspofungin to inhibit fungal growth. The IC50 values of Leishmania strains ranged from 23.02 to 155.80 µg/ml (mean 90.25 ± 39.01 µg/ml), and it was significantly higher (P value = 0.02) than IC50 values of Candida spp. (ranged from 0.001 to 0.12 µg/ml, mean = 0.05 ± 0.05 µg/ml). The reduced growth rate of the parasite was found with exposure to 50 µg/ml of caspofungin. Growth inhibition of Leishmania donovani is significantly lower with caspofungin and could be used to protect the parasite cultivation from fungal contamination.
Highlights
Leishmania parasite cultivation in routine diagnostic and research laboratories is facing a major problem of microbial contamination despite strict adherence on aseptic microbiological practices
Lipid composition in Leishmania parasite cell membrane is similar that in fungi and yeast [4]. ese drugs have been shown to interact with Toll-like receptors and CD14 to induce signal transduction and release of inflammatory cytokines [5]
Results e two Leishmania strains BPK206/0 cl10 and BPK632/0 were tested to determine IC50 against caspofungin, and detailed results were presented in Table 1. e IC50 values of Leishmania strains ranged from 23.02 to 155.80 μg/ml, as shown in Table 1. e parasite growth curve was maintained with different concentrations of caspofungin (10 μg/ml, 25 μg/ml, and 50 μg/ml) along with control
Summary
Leishmania parasite cultivation in routine diagnostic and research laboratories is facing a major problem of microbial contamination despite strict adherence on aseptic microbiological practices. Yeast and yeast-like organisms are always reported as a major source of contamination during parasite culture. The antifungal compounds are rarely used to prevent fungal contamination in parasite culture. Ese drugs have been shown to interact with Toll-like receptors and CD14 to induce signal transduction and release of inflammatory cytokines [5]. These drugs significantly affect parasite’s cell physiology that can disrupt parasite cell cycle. Knowing the fact of antifungal antibiotics that are potential inhibitors to the parasite growth itself, the exploration of antifungal compound which is not inhibiting the parasite growth could prevent the fungal contamination
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