Aims A multidisciplinary approach was used to compare phenolic composition, radical scavenging and antimicrobial activity of propolis samples from different geographical localities, and plant resin against various microorganisms. Methods and Results Using UHPLC-qqqMS quantitative analysis, 28 phenolic compounds were determined. Caffeic and p-coumaric acids were identified as main phenolic acids in poplar propolis samples, except samples from Russia (P6) and China (P7). Radical scavenging activity (applying DPPH spectrophotometric assay) showed the highest activity of Serbian (40 center dot 51%) and Chinese (53 center dot 21%) propolis samples. Broth microdilution method was used for the oral cavity, fungal phytopathogenic and human vaginal isolates which have been identified at a molecular level. The most sensitive bacterial isolates were Lactobacillus acidophilus (MIC of 0 center dot 03-0 center dot 13 mg ml(-1)) and the oral streptococci isolates (MIC values of 0 center dot 19-0 center dot 13 mg ml(-1)). The most sensitive fungal phytopathogenic isolate was Fusarium oxysporum (MIC 0 center dot 003 mg ml(-1)). All samples, except propolis from Serbia (P4) and Turkey (P5), showed a strong antifungal activity against Fusarium sporotrichioides, Fusarium subglutinans and Fusarium proliferatum. Conclusion The results of various tests indicate good radical scavenging and antimicrobial activity against important human and plant pathogens. Significance and Impact of the Study A detailed propolis analysis is important when proposing a preparation of new biological antimicrobial products which have a positive impact on human health and reduce antibacterial resistance.