The aim of the study was to survey the chemical composition, antimicrobial and antioxidant activities of Dittrichia graveolens essential oil in in vitro conditions. GC-FID and GC-MS analyses were performed to evaluate the chemical composition of essential oil. The antimicrobial activity against different kinds of microorganisms was determined by microbroth dilution assay. The antioxidant activity was evaluated by DPPH free radical scavenging system. Studies of chemical composition of essential oil revealed the presence of borneol (38.2%) and bornyl acetate (14.9%) as major constituents. Essential oil showed strong antimicrobial activity against Enterococcus faecalis isolates and Bacillus cereus. Staphylococcus aureus and Escherichia coli are less sensitive to the oil. The strong antimicrobial activity of oil against clinical isolates of bacteria was demonstrated. In addition, D. graveolens oil notably reduced the concentration of DPPH free radical with higher efficacy than that of trolox at 70 min.