In recent years, essential oils has received increasing attention as natural antimicrobials for use against a variety of pathogens, replacing conventional chemical preservatives to achieve a more healthy food. Among a variety of essential oils, the peppermint (Mentha piperita L.) essential oil (PEO), recognized as GRAS (Generally Recognized as Safe) in the dose usually possible to use in foods, appear as promising natural compounds for controlling postharvest decay in fruits, such as the anthracnose caused by Colletotrichum spp. Considering this aspects, the aim of this study was to evaluate the efficacy of PEO to control the mycelial growth of different Colletotrichum species (C. asianum, C. dianesei, C. fructicola, C. tropicale and C. karstii) using an in vitro assay. The effect of the PEOs (Ferquima Ltda) added to the PDA (Potato Dextrose Agar) at a range of concentrations (18.04; 9.02; 4.51; 2.25 µg/mL) on the mycelia growth of target fungal strains during 10 days (28°C) was determined with the poisoned growth substrate technique (MAQBOOL et al.,2011). The mycelial growth was measured (cm/day) and the result expressed as percentage of Mycelial Growth Inhibition (MGI), compared with the control treatment (PDA + 0 µg/mL PEO) and Rate of Mycelial Growth (RMG). The complete inhibition of the target fungi species (fungicidal effect) was observed when the PEO was assayed at concentrations higher than 4.51 µg/mL, while the lowest PEOs concentration tested (2.25 µg/mL) was able to reduce (p<0.05) the RMG (0.67-0.11 cm/day), showing a fungistatic effect, when compared with the control (1.06-0.65 cm/day). These results suggested that PEOs have the potential to control the mycelial growth of different species of the postharvest pathogenic fungus Colletotrichum. Still, these findings support further studies to assess the efficacy of PEO to control the target fungal strains in fruits.