The present study assessed the antimicrobial and anticancer activities of Brachythecium populeum (Hedw.) Bruch, Schimp. & W.Gumbel (Brachytheciaceae) extracts using two different extracting methods. The antibacterial and antifungal activity of seven solvent extracts of Brachythecium populeum was examined against six species of bacteria and seven species of fungi by using well diffusion method. The minimal inhibitory concentration (MIC) of the extracts of acetone and C was determined. Extract C has the highest effect against E. faecalis (13 mm), B. subtilis and S. aureus (12 mm). The inhibition zone was showed by acetone extract against E. faecalis (13 mm), P. aeruginosa (10 mm) and B. subtilis (9 mm). Furthermore, all extracts exhibited different activities on P. aeruginosa. The MIC ranges acetone and C extracts against bacterial strains were from 93.8-375 mu g/mL. Cytotoxic activities of extracts of acetone, A and C of B. populeum (0.17, 1.7, 17, 85 and 170 mu g/mL) were tested against rat glioma cells by using MTT assay, after 24 and 48 h. Extracts of acetone, A and C showed a moderate toxicity, however high concentrations of extract A significantly decreased the survival of glioma cells in 48 h. The study confirms the antimicrobial and cytotoxic activities of B. populeum, extracted using various solvents. It is suggested that the active substances to be obtained by using different solvents from B. populeum may be the active substances of various drugs in the future. Further studies are needed.