Current strategies to control leishmaniasis are mainly based on chemotherapy. However, none of the available drugs can be considered to be ideal to treat this disease. Because of the hydrophobic nature and bioactivities of their components, essential oils (EOs) can be considered as important sources for developing agents against intracellular pathogens, such asLeishmaniaparasites. In this study, we report the chemical characterization, antileishmanial activities, and cytotoxicity effect of the EO fromPluchea carolinensis(Jacq.) G. Don. (Asteraceae). Chemical analysis revealed that EO from aerial part fromP. carolinensisis composed of 44 compounds. The main component was selin-11-en-4α-ol, which made up 51.0%.In vitroantileishmanial studies showed thatP. carolinensisEO inhibited the growth of promastigotes (IC50 = 24.7 ± 7.1 μg/mL) and amastigotes (IC50 = 6.2 ± 0.1 μg/mL) ofLeishmania amazonensis, while cytotoxicity evaluation revealed fivefold higher values than those for the parasites. In a model of experimental cutaneous leishmaniasis in BALB/c mice, five doses of EO at 30 mg/kg by intralesional route demonstrated smaller lesion size and parasite burden (p< 0.05) compared with animals treated with Glucantime® and untreated mice. In conclusion,in vitroandin vivoresults showed the potentialities of EO fromP. carolinensiswith the future possibility of a new alternative in the treatment for leishmaniasis.