Background: The increasing resistance to antifungal drugs and the reduced number of available drugs led to the search for therapeutic alternatives among aromatic plants and their essential oils, empirically used by antifungal effects. OBJECTIVES: The purpose of the current study was to evaluate the antifungal activity of Trachyspermum ammi essential oil (EO) against the most frequent pathogenic fungi including Candida, Aspergillus, Chrysosporium and Trichophyton species. METHODS: EO from the seeds of the plant was obtained by hydrodistillation. Susceptibility tests were expressed as growth inhibition zone (diameter) using disk diffusion method and minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) using broth microdilution method. RESULTS: Results of susceptibility tests showed that T. ammi EO was effective against all the tested strains. The diameters of growth inhibition zone of the EO were between 11 mm and 60 mm. The EO was also the most active, with MIC and MFC values ranging from 0.3 to 2.5 mg/ml and 0.6 to 5 mg/ml, respectively. The EO of T. ammi showed a significant degree of antifungal activity against different Candida species in comparison with other fungi (p<0.05). CONCLUSIONS: The present study indicated that T. ammi EO has considerable antifungal activity, deserving further investigations for its clinical application for treatment of fungal infections.