Several essential oils of medicinal plants possess proven antimicrobial activity and are suitable for applications on the food industry. The oil from Cymbopogon citratus (lemongrass) has been reported to have a wide range of biological activities. However, there are few controlled studies confirming its antimicrobial activity. The aim of our study was to assess the antimicrobial potential of lemongrass oil against a wide spectrum of food spoilage and pathogenic bacteria and yeast in liquid and vapour phase. The chemical composition of the oil was analyzed by gas chromatography; 14 components were identified and geranial (28.93 %), neral (24.30 %) and myrcene (23.92 %) were the most abundant constituents. Lemongrass oil showed potent antimicrobial activity against Gram positive bacteria and exhibited the strongest antifungal effect against Candida albicans and C. parapsilosis. Minimum inhibitory concentration varied from 0.019 to 1.25 mg mL−1 for Gram positive bacteria and yeasts, indicating Staphylococcus aureus, S. epidermidis and C. albicans as the most susceptible strains. Moreover, the diameter of inhibition zone (DIZ) increased with increasing essential oil (EO) volume. Considerably superior antimicrobial activity was observed in the vapour phase. The DIZ resulting from the exposure to EO vapour was significantly larger than that from the same volume in the liquid phase. The DIZ varied from 22 to 90 mm for Candida strains. There is growing evidence that EO in vapour phase are effective antimicrobial systems and that they do have advantages over the use of oil in liquid phase. Our results imply that lemongrass oil could be useful for the development of novel types of natural preservatives for food control.