The stable fly, Stomoxys calcitrans (Linnaeus 1758), is a hematophagous fly responsible for causing loss of performance in horses, causing losses in cattle productivity, and impacting the animals' health through the spread of pathogenic microorganisms. The objective of this work was to investigate the insecticidal activity of essential oil obtained from Melaleuca alternifolia (Cheel), presenting high 1,8-cineole content, against S. calcitrans adults. Insecticidal activity was determined using surface application methods and exposure to oil impregnated paper. It was observed that treatments at 25 and 50 µg/cm² (P < 0.05) present fumigant activity through exposure to the impregnated paper, and in the first 15 min of exposure, the mortality rates obtained for these treatments were, respectively (96.6 ± 3.3% and 100%), equivalent to the positive control. Using the superficial application method, the only treatment concentration presenting adulticidal action was 5% (w/v) (P < 0.05). Respective toxicities LC₅₀ (%, w/v) and LC₈₀ for the impregnated paper method were 1.06 ± 0.02 and 1.47 ± 0.17; for the superficial application method, they were 3.82 ± 0.65 and 5.53 ± 0.74. As demonstrated, M. alternifolia essential oil presents adulticidal potential against S. calcitrans.