The aim of this research was to determine the phytoconstituents of the essential oil (EO) obtained from stems of Croton tricolor and to study its antifungal activity against several Candida species. The EO was extracted from C. tricolor stems by hydrodistillation with a Clevenger apparatus. This EO was characterized by four major components: epiglobulol, α-bisabolol, α-trans-bergamotol and β-caryophyllene. Microbiological assays were performed using the microdilution method in liquid medium to determine the minimum inhibitory concentration (MIC) for each Candida strain. EO concentrations from 1.0 to 1024 μg/mL inhibited the growth of all tested strains. Overall, the microbiological assays showed that C. tricolor EO has significant inhibitory activity against several Candida species. Thus, the C. tricolor stem EO is a promising natural product in the treatment of candidiasis.