Bacterial Susceptibility Outweighs Declared Botanical Origin in Shaping Honey Antimicrobial Activity: An Integrated Biochemical and Microbiological Analysis

<jats:p>Honey antibacterial activity reflects both the properties of the honey matrix and the susceptibility of the target microorganism, yet the relative importance of these sources of variation remains insufficiently resolved. This study investigated whether species-dependent bacterial susceptibility is reflected more consistently in honey antibacterial activity than declared botanical type and whether integration of antimicrobial and biochemical traits reveals botanical-type-associated bioactivity patterns that are not apparent from individual endpoints. Thirty-seven honey samples representing seven declared botanical types from Western Pomerania, Poland, were analysed for total phenolic content, ferric reducing antioxidant power, and minimum inhibitory and bactericidal concentrations against Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Pseudomonas aeruginosa, and Proteus vulgaris. Buckwheat honeys showed the highest observed phenolic content and FRAP activity. Species-dependent differences in MIC and MBC were detected more consistently than differences among declared botanical types; significant botanical-type effects for individual antimicrobial endpoints were observed only against S. aureus. P. vulgaris was generally the most susceptible species, although susceptibility patterns differed between inhibitory and bactericidal endpoints. Associations between biochemical parameters and antimicrobial activity were limited and directionally inconsistent. In contrast, integration of five MIC variables, five MBC variables, FRAP activity, and total phenolic content revealed significant botanical-type-associated structuring of the overall bioactivity profile. This effect remained significant after exclusion of botanical types represented by single samples (pseudo-F = 2.66, R2 = 0.283, p = 0.0001). These findings indicate that botanical classification alone has limited value for predicting antibacterial activity against a particular microorganism, whereas combined species-resolved microbiological and biochemical profiling provides a more informative framework for characterising honey bioactivity and deriving descriptive bioactivity fingerprints.</jats:p>