Mushrooms are the most widespread organisms on Earth with a significant role in the ecological processes in numerous, especially forest, ecosystems. The aim of this study was to investigate in vitro the antimicrobial activity of Macrolepiotia procera, Lactarius piperatus, Hydnum repandum, Clitocybe nebularis and Craterellus cornucopioides collected in the forests of the Primorsko-goranska county. Methanol extracts of the selected mushrooms were tested against two gram negative (Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853) and two gram-positive bacteria (Staphylococcus aureus ATCC 29213, Enterococcus faecalis ATCC 292129). The antimicrobial effect was determined using diffusion methods (disc diffusion and diffusion in the wells) and a microdilution method which determines minimal inhibitory and minimal bactericidal concentrations. At the highest concentration tested (200 mg/mL), all fungi showed an inhibitory effect on all selected bacteria, except M. procera which did not inhibit the growth of bacterium E. faecalis evan at the highest concentration. M. procera showed the weakest antimicrobial activity compared to other investigated mushrooms. C. nebularis extract has modest antimicrobial efficacy, especially against gram-positive bacteria. The best antimicrobial effect was observed for fungi C. cornucopioides and H. repandum, which at concentrations of 100 mg/mL acted bacteriostatic against all selected bacteria. Among the gram-positive bacteria, S. aureus proved to be more susceptible to mushrooms extract, while among gram-negative more susceptible was P. aeruginosa. E. faecalis was the most resistant among the bacteria tested. 41 The results have shown that edible mushroom extracts, especially C. cornucopioides and H. repandum with expressed antimicrobial effects, can be the source of new bioactive compounds with potential use in medicine, as well as pharmaceutical and food industry.