In a recent study, our research group demonstrated that the essential oil of Ocotea odorifera (EOOO)and its major compound safrole potentiated the action fluoroquinolones, modulating bacterial resistancepossibly due to direct inhibition of efflux pumps. Thus, in the present study, we investigated whether thesetreatments could enhance the activity of gentamicin and erythromycin against multidrug-resistant (MDR)bacteria. The EOOO was extracted by hydrodistillation, and the phytochemical analysis was performed bygas chromatography coupled to mass spectrometry (GC–MS). The antibiotic-enhancing effect of the EOOOand safrole against MDR strains of Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa wasanalyzed by the broth microdilution method. The chemical analysis confirmed the presence of safrole as amajor component among the 16 compounds identified in the EOOO. Both the essential oil and the isolatedcompound showed clinically relevant antibacterial activities against S. aureus. Regarding the modulationof antibiotic resistance, the EOOO was found to enhance the activity of erythromycin against the strains ofP. aeruginosa and S. aureus, as well as improving the action of gentamicin against S. aureus. On the otherhand, safrole potentiated the activity of gentamicin against the S. aureus strain alone. It is concluded,therefore, that the EOOO and safrole can enhance the activity of macrolides and aminoglycosides, andas such are useful in the development of therapeutic tools to combat bacterial resistance against theseclasses of antibiotics.