The new buzz: Investigating the antimicrobial interactions between bioactive compounds found in South African propolis

Ethnophannacological relevance: Propolis, a resinous substance produced by the Apis mellifera bee, contains a number of flavonoids sourced from plants found in the surrounding region. Whilst bees use this substance to seal off and protect the beehive, humans have used propolis therapeutically for centuries, making use of its antibacterial, antiseptic, antipyretic and wound healing properties, among others. South African propolis is rich in the flavonoids pinocembrin, galangin, and chrysin and very little previous research has been conducted on the antimicrobial effects of these compounds. Aim of the study: To obtain an understanding of the antimicrobial activity of the compounds pinocembrin, galangin, and chrysin, both independently and in combination. Materials and methods: The compounds pinocembrin, galangin and chrysin were investigated for interactive antimicrobial activity by determining the minimum inhibitory concentrations (MIC), minimum bactericidal concentrations (MBC), anti-quorum sensing activity, biofilm studies, and toxicity studies (brine shrimp lethality assay). Results: Minimum inhibitory concentration results demonstrated that combinations of compounds showed better inhibitory activity than single compounds. When the flavonoids were tested in combination using the MIC assay, synergy was noted for 22% of the 1:1 ratio combinations and for 66% of the triple 1:1:1 ratio combinations. Similarly, MBC results showed bactericidal activity from selected combinations, while the compounds on their own demonstrated no cidal activity. Quorum sensing studies showed that compound combinations are more effective at inhibiting bacterial communication than the individual compounds. Biofilm assays showed that the highest percentage inhibition was observed for the triple combination against E. coli at 24 h. Finally, brine shrimp lethality studies revealed that combinations of the three compounds had reduced cytotoxicity when compared to the individual compounds. Conclusion: The results obtained in this study demonstrate that the compounds found in South African propolis work synergistically to achieve an optimal antimicrobial effect, whilst simultaneously minimizing cytotoxicity.