Introduction: In different branches, good hygiene practices require proper cleaning and additional disinfection of work surfaces and devices. Cleaning removes visible organic and inorganic impurities from surfaces, while disinfection removes most or all microorganisms present on the surfaces, except spores. Certain disinfectants contain compounds that can be harmful to human health and the environment. Therefore, alternative products such as vinegar are increasingly being used. Vinegar is produced by the fermentation of alcohol and alcoholic beverages. The main ingredient of vinegar is monocarboxyl acetic acid, but other substances are also present, depending on the type of fermentation. According to definition, an antimicrobial agent is considered effective if it reduces the microbial count by at least 5 log10 in 5 to 10 minutes or by at least 3 log10 in 30 seconds. Purpose of the work: To investigate the inhibitory effect exerted by commercially available vinegars and acetic acid on different Gram positive and negative bacteria, and fungi. Methods: In order to study the effect of vinegars and acetic acid on standard bacterial and fungal strains, the following methods were used: disk diffusion assay, dilution method, effect of vinegars on die-off time curve for microorganisms in suspension (suspension assay), surface assay with vinegar action on microorganisms on surfaces, and rinsing of vinegars (membrane filtration) after their action on microorganisms. Results: Vinegars and acetic acid were the most effective against Microccocus luteus. The most resistant were Escherichia coli, Staphylococcus aureus and Bacillus cereus. On average across all methods, 9% alcohol vinegar was the most effective, and 1% acetic acid was the least effective. Discussion and conclusion: The test microorganisms used demonstrated varying susceptibility to vinegars and acetic acid. In almost all methods and against the test microorganisms used, vinegars and acetic acid showed bacteriostatic, and only rarely bactericidal action. In most cases, antimicrobial effect depended on the vinegar’s nominal acidity level.