Honey is an inexpensive, food-based option for treating diabetic foot ulcers (DFUs) due to its anti-inflammatory and antibacterial properties. It has proven effective against bacterial biofilms and multidrug-resistant bacteria, positioning honey as a promising candidate for DFU management. However, honey’s physicochemical properties and concentration variations can lead to differing bacterial responses. This study aimed to assess the effects of various honey types and concentrations on bacterial biofilms. Seven honey types were tested at concentrations of 1x, 5x, and 10x the minimum inhibitory concentration (MIC) against biofilms of Staphylococcus aureus, Escherichia. coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Candida albicans.

Results indicated that honey types 3 and 4 achieved significantly higher biofilm removal for C. albicans compared to types 5 (p = 0.008; p = 0.001), 8 (p = 0.022; p = 0.003), and 9 (p = 0.009; p = 0.001) at the same concentrations. Similarly, honey types 2 and 4 were more effective against E. coli than type 9 (p = 0.016; p = 0.004). Higher honey concentrations resulted in greater biofilm removal for C. albicans and E. coli (p = 0.004, both), and more pronounced biofilm metabolic inactivation for C. albicans, K. pneumoniae (p = 0.004, both), and P. aeruginosa (p = 0.048). Principal Component Analysis suggested a correlation between pollen content and antimicrobial activity. Overall, honey demonstrated significant potential in removing bacterial biofilms and inhibiting metabolic activity, particularly at higher concentrations. The differences in bacterial responses to honey treatments may be attributed to variations in the honeys' physicochemical properties and bacterial strain sensitivity.