Staphylococcus aureus causing mastitis in cows reduces the quality of milk and dairy products leading to huge economic losses. Treatment of this disease with conventional antibiotics has problems as drug-resistant pathogens; as a consequence, there is a growing need to use natural products as potential antimicrobial agents. The present study was conducted to investigate the antimicrobial potential of essential oils (EOs), and their synergistic combinations against S. aureus from Subclinical Bovine Mastitis obtained in the State of São Paulo – Brazil. In this experimental study, nine different strains were selected, and the bacterial suspensions were inoculated in a final concentration of 107 CFU.mL-1 to perform the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of three different EOs (clove - Eugenia caryophyllus, arrayan - Luma chequeen, and spearmint - Mentha spicata). The fractional inhibitory concentration (FIC) index for the synergism of their possible combinations, were investigated. Chemical compositions of EOs were analyzed by gas chromatography coupled to a mass selective detector (GC/MSD). The MIC and MBC values ranged 0.39 – 1.10 mg.mL-1 and for 0.66 – 2,20 mg.mL-1 for clove, 1.12- 4.48 mg.mL-1 and 2.99 – 14.93 mg.mL-1 for arrayan, and 1,35 – 18.56 mg.mL-1 and 7.73 – 18.56 mg.mL-1 for spearmint, respectively. GC/MSD analysis identified the main constituents of clove, arrayan, and spearmint EOs, which were eugenol (86.25%), limonene (32.64%), and carvone (63.21%), respectively. Although cloves obtained better MIC values when analyzed alone, the best combination was spearmint + arrayan, which displayed a synergistic effect against S. aureus by reducing the initial MIC values. These results indicate that the EOs analyzed alone and in synergistic combination along with their major components can influence the antimicrobial efficacy against the pathogen, suggesting that these compounds can be considered alternative antistaphylococcal agents for subsequent use in treating mastitis in dairy cows.