Many plants contain active substances that are known to be effective in both enhancing the wound healing process and lowering the incidence of wound infections. Previous studies have shown that bryophytes produce a variety of secondary metabolites that present pharmaceutical activities including antimicrobial activity against various pathogenic bacteria and fungi. The aim of this study was to investigate the antimicrobial activity of Hedwigia ciliata var. leucophaea (He) against 17 bacterial and 1 fungal strains and analyse the composition by GC/MS. He samples were collected from Akdag Mountain (Amasya, TURKEY) and subjected to ethanol extraction after air drying. The in vitro antimicrobial activities of He extracts having 9 mg.mL-1 concentrations were assessed against a wide range of strains by disk diffusion method and the results were supported by a MIC (minimum inhibitory concentration) test. Our present study has shown that the ethanol extract of He has antimicrobial activity against several Gram positive and Gram negative microorganisms tested, but its antimicrobial activity is notable especially against B. subtilis, E. faecalis, S. camosus, and S. epidermidis. These results are the very first report of the antimicrobial activity of He and its.composition. The results obtained herein indicate that ethanol extract of He contains several active metabolites and it is active against several microorganisms. But further researches, especially cytotoxicity and genotoxicity tests are needed to be conducted to conclude whether He extracts can be used safely in terms of their antimicrobial activity.