The aim of present study was to compare the antimicrobial and antioxidant activity of Thymus spathulifolius on the basis of the chemical composition of oil obtained by hydrodistillation. The essential oil was analyzed by GC and GC-MS and 29 components, representing 99.9 % of the essential oil, were identified. Carvacrol (49.0 %), thymol (17.8 %) and p-cymene (12.3 %) were the main components. The antimicrobial activity of the essential oil was assessed by the disc diffusion and microbroth dilution method. All tested microorganisms were inhibited by essential oil sample. The strongest activity was observed against Escherichia coli, Enterobacter aerogenes and Pseudomonas aeruginosa with MIC values of 125 mu g/ml. The essential oil is also active on test fungi. A. flavus seemed to be sensitive to the investigated oil with MIC values of 250 mu g/ml. The antioxidant activity of the essential oil (100-1000 mu g/ml) was determined by means of the DPPH radical-scavenging method. At 1000 mu g/ml the essential oil concentration of T. spathulifolius, 77.31 +/- 0.21% DPPH was scavenging.