“…Axenic amastigote forms of L. infantum were cultured as described in the literature. 35 The promastigotes were transformed into amastigotes by culturing for three days in BHI medium supplemented with 10% heat-inactivated fetal calf serum, 1 g L −1 β-alanine, 100 mg L −1 l -asparagine, 200 mg L −1 sucrose, 50 mg L −1 sodium pyruvate, 320 mg L −1 malic acid, 40 mg L −1 fumaric acid, 70 mg L −1 succinic acid, 200 mg L −1 α-ketoglutaric acid, 300 mg L −1 citric acid, 1.1 g L −1 sodium S9 bicarbonate, 5 g L −1 2-[morpholino]ethanesulfonic acid, 0.4 mg L −1 hemin and 10 mg L −1 gentamicin at a pH of 5.4 at 37 °C. The screening was performed in 96-well microtiter plates maintained at 37 °C.…”