Amastigotes from L(L)amazonensis(La); L(L)venezuelensis(Lv); L(V)brasiliensis(Lb) and L(L)chagasi(Lch) were cultured in a liquid culture medium without mammalian cells present (O'Daly's medium). 132 patients from a hyperendemic region (prevalence 24.8 o/oo) had ulcers, scars and nodules, more frequent in legs and arms after cutaneous leishmaniasis (CL) infection. Time for remission of lesions were: Spontaneous remission (SR): 7 weeks; Glucantime®: 9 weeks; vaccination with La, Lv, Lb, Lch amastigotes TLCK treated and NP-40 extracted (VT): 7 weeks; with autoclaved L(L)mexicana promastigotes +BCG : 18 weeks. Delayed type hypersensitivity response (DTH) response with polyvalent and monovalent leishmanine was higher in CL patients than healthy controls (p0.05) and increased in active secondary vs. primary infection (p0.0001) with diagnostic value 1.74 for active infection and 1.81 post-clinical remission of lesions.