300 mg/m2 per day for five days.

Promastigotes of L. donovani (1 107 parasites/mL) were cultivated in TC-199 medium supplemented with 20 % of IFCS, at which previously 38 μM of the edelfosine or miltefosine or ilmofosine had been added. After five days, the parasites were spun to 2,000 g for 10 minutes, and the pellet was washed in PBS and fixed for two hours in cacodylate buffer (0.05 M, pH 7.4) containing p-formaldehyde and glutaraldehyde (in equal shares) 2 % (v/v). Then, it was transferred for eight hours in 0.1 M cacodylate buffer with 2 % of glutaraldehyde. Postfixation was carried out for two hours with 2 % of osmium tetroxide (p/v) and potassium ferrocyanide 2 % (p/v). Once the fixation was completed, the staining of the blocks was performed with 1 % of uranyl acetate solution (p/v) for 30 minutes. Then, the blocks were dehydrated in a growing series of alcohols. The infiltration was carried out in resin Spurr for one hour and the polymerization was done in 12 hours