The report of a new autochthonous case of human American Visceral Leishmaniasis (AVL) in 1992 in the village of Guayabita, Aragua State, Venezuela (10 degrees 16'N, 67 degrees 28'W; 500 m asl), led us to undertake an epidemiological study in this locality. A demographic survey was conducted using a structured questionnaire in which data was collected regarding sex, age, occupation, length of residence in the area and migratory history. A leishmanin skin test (LST) was applied and samples of venous blood for counter immunoelectrophoresis (CIEP) and immunofluorescent antibodies (IFAT) tests were drawn. The prevalence of positive LST was 11.4%. The positivity was highest among males and increased with age. Young males seemed to be more exposed to infection than females. Since occupation per se did not seem to account for this association, it may be explained as a gender-associated behavior, leading to different degrees of exposure to sand flies. The canine population was also screened for leishmanial infection. One dog was shown to be parasitologically infected with Leishmania sp. Four out of 71 dogs (5.6%) were positive for FG, CIEP, IFAT and Western Blot and 11 (15.5%) were positive for CIEP. These results suggest that after almost 30 years of epidemiological silence, American visceral leishmaniasis has re-emerged in this focus.