Aims The study was focused on the evaluation of antimicrobial activity in vitro of the essential oil (EO) of leaves from Schinus molle against bacteria and fungi of clinical importance in the search for the discovery of new active compounds. Methods and Results The chemical composition of the S. molle EO was determined by gas chromatography/mass spectrometry and its antimicrobial effect was verified by broth microdilution method. The major compounds found were beta-pinene (25.23%), epi-alpha-cadinol (21.29%), alpha-pinene (18.72%), myrcene (11.54%) and sabinene (5.02%). The EO showed significant antifungal activity against Paracoccidioides brasiliensis (39.06 mu g ml(-1)), weak action against Cryptococcus neoformans (625 mu g ml(-1)) and Trichophyton quinckeanum (625 mu g ml(-1)) and was inactive against Candida sp. In the analysis of the antibacterial action, the micro-organisms tested did not show sensitivity. Conclusions This study showed a promising result of S. molle volatiles against the fungus P. brasiliensis, which causes paracoccidioidomycosis (PCM), a systemic mycosis of great clinical importance in Latin America. Significance and Impact of the Study The results found here are novel and encourage investigations of the compounds present in this EO, which represents a source of molecules with potential use in the treatment of PCM.