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The aim of this study was to investigate the chemical composition of Origanum vulgare L. essential oil, the inhibitory effect of the oil on the cell viability of Staphylococcus aureus strains isolated from foods, and the influence of sub-inhibitory concentrations of the oil on some physiological attributes of these strains. GC–MS analysis showed that carvacrol (57.71%) was the most prevalent compound in the oil, followed by p-cymene (10.91%), γ-terpinene (7.18%), terpinen-4-ol (6.68%) and thymol (3.83%). The results showed that O. vulgare essential oil at 0.03, 0.6 and 0.12μLmL⁻¹ inhibited the cell viability of Staph. aureus. At 0.12μLmL⁻¹ the oil caused cidal effect with decrease ≥3log cycles of the initial inoculum after 15min of exposure. Sub-inhibitory concentrations (0.03 and 0.015μLmL⁻¹) of the oil suppressed some physiological attributes of the Staph. aureus strains such as coagulase, lipase and salt tolerance …