The composition of the essential oil from aerial parts of Thymbra spicata L. from Turkey was analysed by GC-MS and its in vitro antimicrobial activity was examined. GC-MS analysis of the essential oil resulted in the identification of twenty-three constituents, representing 97.04% of the oil. The major compounds detected in the essential oil were carvacrol (60.39%), gamma-terpinene (12.95%), and p-cymene (9.61%). The in vitro efficacy of the essential oil against 21 bacteria and seven Candida species was examined using disc diffusion and minimum inhibitory concentration (MIC) methods. The essential oil demonstrated strong anti-microbial activity in a wide spectrum against most microorganisms, particularly the yeasts tested. This is the first report on the anticandidal properties of the essential oil of T. spicata. In conclusion, this study confirms that T. spicata essential oil could be considered as a natural antimicrobial source.