We compared the clinical findings and diagnostic methods for 66 patients with cutaneous leishmaniasis in the state of Bahia, Brazil, who were infected by Leishmania (Viannia) braziliensis (group A), with those for 68 patients in the state of Amazonas, Brazil, who were mainly infected by Leishmania (Viannia) guyanensis (group B). Differences were observed with regard to number, size, and location of skin lesions and to the pattern of lymphatic involvement. Patients in group B had smaller and more numerous lesions, which were frequently located above the waist, versus the larger but less numerous lesions among patients in group A, which were usually located on the lower limbs. Lymphatic involvement was present in 55 (83.3%) of the 66 patients in group A and in 42 (61.8%) of the 68 patients in group B (P=0.005). The positivity rates of imprints and skin culture procedures were higher in group B. Sensitivity of in vitro culture of skin aspirates was 47.0% and 91.2% for groups A and B, respectively (P<.001). Although hamster inoculation showed similar results in both groups, the interval before development of disease was shorter in group B. Our data provide substantial evidence that indicate that the disease caused by these species differs with regard to clinical presentation and diagnostic approach.