| Summary: | <i>Staphylococcus aureus</i> is still one of the leading causes of both hospital- and community-acquired infections. Due to the very high percentage of drug-resistant strains, the participation of drug-tolerant biofilms in pathological changes, and thus the limited number of effective antibiotics, there is an urgent need to search for alternative methods of prevention or treatment for <i>S. aureus</i> infections. In the present study, biochemically characterized (HPLC/UPLC–QTOF–MS) acetonic, ethanolic, and water extracts from fruits and bark of <i>Viburnum opulus</i> L. were tested in vitro as diet additives that potentially prevent staphylococcal infections. The impacts of <i>V. opulus</i> extracts on sortase A (SrtA) activity (Fluorimetric Assay), staphylococcal protein A (SpA) expression (FITC-labelled specific antibodies), the lipid composition of bacterial cell membranes (LC-MS/MS, GC/MS), and biofilm formation (LIVE/DEAD BacLight) were assessed. The cytotoxicity of <i>V. opulus</i> extracts to the human fibroblast line HFF-1 was also tested (MTT reduction). <i>V. opulus</i> extracts strongly inhibited SrtA activity and SpA expression, caused modifications of <i>S. aureus</i> cell membrane, limited biofilm formation by staphylococci, and were non-cytotoxic. Therefore, they have pro-health potential. Nevertheless, their usefulness as diet supplements that are beneficial for the prevention of staphylococcal infections should be confirmed in animal models in the future.
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