The search for natural sources to control microorganisms is of interest in food production. This study evaluated the chemical composition and antimicrobial activity of the essential oil from Psidium cattleianum leaves. The essential oil was extracted by hydrodistillation, and identified by GC-MS. The predominant class of compounds was sesquiterpenes (47.6%) and the major compounds were trans-β-caryophyllene (14.7%), 1,8-cineole (11.7%) and ɣ-muurolene (5.6%). The antimicrobial activity was carried out by microdillution technique against eight fungi and eight bacteria. The minimum inhibitory concentration ranged from 0.17 to 11.25 mg mL-1 for fungi, and from 1.40 to 16.87 mg mL-1 for bacteria. The highest activities were against fungi Aspergillus fumigatus (ATCC 1022), Aspergillus ochraceus (ATCC 12066), Aspergillus versicolor (ATCC 11730), and Trichoderma viride (IAM 5061), and bacteria Pseudomonas aeruginosa (ATCC 27853), Bacillus cereus (clinical isolate), and Staphylococcus aureus (ATCC 6538) with potential to prevent foodborne diseases.