Abstract The chemical composition and antimicrobial activity of the hydrodistilled essential oil isolated from the aerial parts of Stachys milanii PetroviÄ, a species endemic to central and eastern regions of the Balkan Peninsula, were examined for the frst time. Fourteen compounds, predominantly monoterpenes, representing 92.2% of the total oil, were identifed, with borneol (49.5%) and terpinen-4-ol (13.2%) as major components. The antimicrobial activity of the isolated oil against Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella enteritidis, Escherichia coli, Staphylococcus aureus and Aspergillus niger was investigated by disc diffusion method and was a moderate one. Key Word Index: Stachys milanii Lamiaceaeessential oil compositionborneolterpinen-4-olantimicrobial activity