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<i>Staphylococcus aureus</i> infections are prevalent in healthcare and community environments. Methicillin-resistant <i>S. aureus</i> is catalogued as a superbug of high priority among the pathogens. This Gram-positive coccus can form biofilms and produce toxins, leading to persistent infection and antibiotic resistance. Limited effective antibiotics have encouraged the development of innovative strategies, with a particular emphasis on resistance mechanisms and/or virulence factors. Medicinal aromatic plants have emerged as promising alternative sources. This study investigated the antimicrobial, antibiofilm, and antihemolysis properties of three different chemotypes of <i>Lippia origanoides</i> essential oil (EO) against susceptible and drug-resistant <i>S. aureus</i> strains. The chemical composition of the EO was analyzed using GC-MS, revealing high monoterpene concentrations, with carvacrol and thymol as the major components in two of the chemotypes. The third chemotype consisted mainly of the sesquiterpene β-caryophyllene. The MIC values for the two monoterpene chemotypes ranged from 62.5 to 500 µg/mL for all strains, whereas the sesquiterpene chemotype showed activity against seven strains at concentrations of 125–500 µg/mL, which is the first report of its anti-<i>S. aureus</i> activity. The phenolic chemotypes inhibited biofilm formation in seven <i>S. aureus</i> strains, whereas the sesquiterpene chemotype only inhibited biofilm formation in four strains. In addition, phenolic chemotypes displayed antihemolysis activity, with IC₅₀ values ranging from 58.9 ± 3.8 to 128.3 ± 9.2 µg/mL. Our study highlights the importance of <i>L. origanoides</i> EO from the Yucatan Peninsula, which has the potential for the development of anti-<i>S. aureus</i> agents.