Although the proven positive effects of propolis on health have been known for many years, research on this topic is not decreasing even today. The bioactive components of propolis consist mostly of polyphenolic components (flavonoids, phenolic acids and acid derivatives) and other organic compounds present in smaller quantities. Variability in the chemical composition, which is the result of variability in propolis source, geographical origin and extraction methods are the main factors that make it difficult to standardize the production technology of propolis based products. Alcohol tinctures of propolis are mostly available on the market, and with the aim of expanding the range of products and availability for the wider population, research is focused on the use of other solvents (vegetable oils, water, glycerol, natural eutectic solvents) and their combinations as a substitute for ethanol. This research was conducted to evaluate the antimicrobial effect of propolis extracts against foodborne pathogens, which can also be found in human microbiology (wound, ear, eye swabs). Alcoholic (80 % ethanol) and non-alcoholic (glycerol:water, 95:5) propolis extracts were prepared by ultrasound-assisted extraction, and the total phenolic content in extracts was determined. After that, the antimicrobial activity of the extracts was tested on four clinical strains isolated from biological samples (Escherichia coli, Salmonella sp., Staphylococcus aureus and Enterococcus faecalis) using the agar diffusion method. The total phenolic content in the non-alcoholic extract of propolis was significantly lower compared to the alcoholic extract, but both extracts showed a moderate inhibitory effect against gram-positive bacteria S. aureus and E. faecalis. Since the test showed that the negative control (glycerol:water) had no effect on the tested bacteria, the antimicrobial activity of the non-alcoholic propolis extract originates from the propolis components and is the motivation for further research into the antimicrobial effect of non-alcoholic propolis extracts on various microorganisms and the connection with the chemical composition and techniques extractions.