Histoplasmosis, a granulomatous disease caused by Histoplasma capsulatum, is endemic in Venezuela. We conducted the current study to appraise retrospectively the demographic data, clinical features, diagnostic methods, and treatment of patients with histoplasmosis from January 2000 to December 2005. We reviewed the medical records of outpatient cases with a diagnosis of histoplasmosis and considered clinical samples processed at our laboratory. We collected demographic, epidemiologic, and clinical data from each case as available, including results of any mycologic examinations performed. Treatment and outcome data were available for some patients. We assessed 158 cases of histoplasmosis: 103 (65.2%) patients came from the Caracas metropolitan area; 53 were associated with acquired immunodeficiency syndrome (AIDS), 14 with tuberculosis, and 8 with paracoccidioidomycosis. Six pediatric patients were malnourished. Epidemiologic data suggested histoplasmosis in most cases. Patients received treatment with itraconazole and/or amphotericin B. Our results may reflect changes in the epidemiology occurring in Venezuela, perhaps due to environmental changes and the presence of AIDS. Several mycologic exams are necessary to ensure a proper diagnosis. More reliable data and statistics on this infection are necessary to monitor outbreaks closely and to establish if there is an epidemic pattern.