The chemical composition of the essential oils obtained by hydrodistillation from the aerial parts of Thymus serpyllum and Thymus serpyllum‘Aureus’ has been investigated by gas chromatography-mass spectrometry (GC-MS). Forty-seven compounds (99.67% of the total oil) wereidentified in the essential oil of T. serpyllum. The main components found in the oil were carvacrol (37.49%), -terpinene (10.79%), -caryophyllene (6.51%), p-cymene (6.06%), (E)--ocimene (4.63%) and -bisabolene (4.51%). Similarly, carvacrol (44.93%), -terpinene(10.08%), p-cymene (7.39%) and -caryophyllene (6.77%) dominated in the oil of T. serpyllum ‘Aureus’. A total of forty three compounds wereidentified in this oil, representing 99.49% of the total oil content. On the basis of the obtained data it was proved that the content of 1-octen-3-ol,eucalyptol, (Z)--ocimene, (E)--ocimene, -terpinene, carvacrol methyl ether, germacrene D and -bisabolene was significantly higher for T.serpyllum while T. serpyllum ‘Aureus’ was characterized by a significantly higher content of 3-octanone, 3-octanol, p-cymene, borneol andcarvacrol. The isolated essential oils were evaluated for their antimicrobial activity against nine reference strains (Escherichia coli, Staphylococcusaureus, Staphylococcus epidermidis, Streptococcus agalactiae, Enterococcus faecalis, Bacillus cereus, Micrococcus luteus, Proteus vulgaris and Candidaalbicans) by the microdilution technique. Based on this test, the minimum inhibitory concentrations (MIC) of essential oil were calculated. Thevolatile oil obtained from T. serpyllum showed the highest antimicrobial activity relative to the strain of E. coli (MIC=0.025 μL/mL) and to theyeast C. albicans (MIC=0.05 μL/mL). Similarly, a significant antimicrobial activity exhibited T. serpyllum ‘Aureus’ essential oil, although the MICvalues obtained in that case for E. coli and C. albicans strains were twice as high and were respectively 0.05 μL/mL and 0.1 μL/mL.