The present work describes the chemical composition and antimicrobial activity of the essential oil of Myosotis alpestris F.W. Schmidt subsp. alpestris (Boraginaceae). The essential oil was obtained from all parts of the plant by microwave distillation and analyzed by GC-FID and GC-MS. Forty-seven components representing 88.2 % of the total oil were characterized and the main components of this species were found to be n-nonanal (11.8 %), decanal (10.8 %), n-octanal (10.7 To), hexahydrofanesyl acetone (6.6 %), o-cymene (3.9 %), tetradecanal (3.7 %), eicosanol (2.2 %) and E-beta-farnesene (2.0 %). The isolated essential oils of M. alpestris subsp. alpestris was tested for antimicrobial activity against the bacteria Escherichia coli, Yersinia pseudotuberculosis, Pseudomonas aeruginosa, Staphylococcus aureus, Enterococcus faecalis, Listeria monocytogenes, Bacillus cereus, Mycobacterium smegmatis and the fungus Candida albicans and Saccharomyces cerevisiae at maximum essential oil concentration in hexane of 1000 mu g/mL and they showed only antibacterial activity against fungus bacteria.