The essential oil (EO) obtained from the leaves of the thyme (Thymus vulgaris L.) grown in Vietnam was found to contain thymol (39.79%), cymene (17.33%), and γ-terpinene (13.45%) as the main volatile components. The antimicrobial activities of this oil were screened against several food-borne bacteria and fungi species. Significant growth inhibition effects against food-borne bacteria Escherichia coli, Staphylococcus aureus, Bacillus cereus, and Salmonella Typhimurium were observed using the standard disc diffusion method. Thyme EO showed the antibacterial effect against all the test pathogenic strains with the inhibition zones were 23.3 ± 0.4 mm, 24.7 ± 0.4 mm, 29.0 ± 0.7 mm, 32 ± 0.7 mm in diameter against B. subtilis, E. coli, S. Typhimurium and S. aureus, respectively. The minimum inhibitory concentration (MIC) determined by micro-dilution method in MHB liquid medium was 1.56 µl/ml. The bactericidal concentrations (MBC) was 3.13 µl/ml for three isolates from B. subtilis, E. coli, and S. aureus, while the MBC tested for S. typhi was 1.56 µl/ml. The antifungal properties of the thyme EO were also determined in this study against three important pathogenic fungi such as Candida albicans, Rhizoctonia solani and Fusarium oxysporum with the inhibition zones ranging approximately from 23.20 ± 0.06 to 44.10 ± 0.03 mm. On the other hands, the results also showed the antioxidant activity of Vietnamese thyme EO and suggested that thyme EO can be applied in food industries as natural flavoring preservatives/additives to control food spoilage and food born bacteria and fungi.