For a long period of time, saturated quaternary ammonium salts have been used as components of compositions that have antimicrobial properties against certain strains of microorganisms. The disadvantageof using such compositions is the formation of resistance of some microorganisms to the latter. Thus, it seems relevant to create antimicrobial compositions based on components with which microorganisms had no contact in the environment. This article presents the results of experimental studies of the antimicrobial activity of synthesized acetylenic quaternaryammonium salts, salts of tertiary amines, as well as commercially available saturated quaternary ammonium salts in relation to test cultures of Proteus mirabilis, Proteus vulgaris, Salmonella typhimurium, Pseudomonas aeruginosa, Escherichia coli, Shigella sonne, Serratia marcescens, Staphylococcus aureus, Candida albicans, Saccharomyces cerevisiae, Penicillium spp. and Bacillus cereus. The diameter of the zones of inhibition of growth by test-cultures of microorganisms was determined by the disk-diffusion method. The action of solutions of acetylenic quaternary ammonium salts of various concentrations was carried out by the method of serial dilutions.The minimum inhibitory concentration of solutions of acetylenic quaternary ammonium salts has been determined. The research results showed that the range of the minimum inhibitory concentration of tri-n-octylprop-2-ynylammonium bromide for the studied test cultures of microorganisms is from 0.1 to 0.0001%, and bromides of tri-n-octylbut-2-ynyl- and tri-n-octyloct -2-ynylammonium - from 1 to 0.0001%, while limiting quaternary ammonium salts exhibit an antimicrobial effect on similar cultures of pathogenic microorganisms at concentrations an order of magnitude higher and more. Thus, solutions of acetylenic quaternary ammonium salts can be promising antimicrobial agents.