Storage-induced chemical changes in active components of honey de-regulate its antibacterial activity

To elucidate reasons for the observed variability in the antibacterial activity of honeys, we analysed a causal relationship between (a) honey floral sources and the activity and (b) the effect of honey storage on stability of compounds conferring this activity. Honeys from diverse floral sources were screened against Escherichia coli (ATCC 14948) and Bacillus subtilis (ATCC 6633) using the broth microdilution method. Among “active” honeys, 37% originated from buckwheat, 18% from clover and 12% from blueberry, indicating that these floral sources produced phytochemical(s) that inhibited bacterial growth. The stability of the putative phytochemical(s) was analysed in “active” honeys (MIC90 6.25% v/v) by measuring the activity every 3–6 months for a period of 1–3 years. A sharp decline in activity against both bacteria was observed in the first 3–6 months of storage. The decline coincided with major changes in chemical composition of honeys which included a significant change in colour (p < 0.0025), extremely significant change in concentration of UV-absorbing compounds (p < 0. 0001) and appearance of melanoidins. While these changes reduced E. coli sensitivity to honey, it rendered B. subtilis completely insensitive. Thus, the data indicates that the presence of phytochemical(s) conferring the antibacterial activity is sensitive to storage. The de-regulation of the antibacterial activity with the concomitant appearance of melanoidins suggests that the active phytochemical components might be sequestered into melanoidin aggregates, losing their function.