Partial purification of amastigotes from cutaneous lesions of American leishmaniasis

Amastigotes of Leishmania mexicana, L. mexicana amazonensis, L. brasiliensis, and L. enriettii were isolated from lesions in infected animals. Numbers of amastigotes recovered ranged from 1 X 10(7) to 7 X 10(8), depending on the strain of leishmania. Trypsinization disassociated the lesions and released the parasites. After 18 to 24 hr incubation at 37 C in tissue culture media with antibiotics, many of the intact host cells attached to the flask. Amastigotes were collected from the media in relatively pure preparations. Electron microscopy revealed no morphological alterations of the amastigotes and minimal contamination by membranes and cell fragments.