Melissopalynology, antioxidant capacity and mineral and toxic element contents were analyzed in eight types of Hungarian honeys. Based on color, two groups were distinguished: light honeys comprised acacia, amorpha, phacelia and linden honeys; while dark honeys included sunflower, chestnut, fennel and sage honeys, with 100 to 300 and 700 to 1500 mAU, respectively. The unifloral origin of each sample was supported using pollen analysis. The absorbance of honey correlated positively with antioxidant capacity determined by three different methods (TRC, DPPH, ORAC), and also with mineral content. The exception was the light amber linden honey with significantly higher K content and antiradical activity than other light honeys. The Mn, Zn and Fe contents were the highest in chestnut, sunflower and fennel honeys, respectively. The black meadow sage honey performed best regarding the content of other elements and antioxidant activity. The concentrations of several toxic elements were below the detection limit in the samples, indicating their good quality. The principal component analysis (PCA) revealed correlations between different antioxidant assays and minerals, and furthermore, confirmed the botanical authentication of the honeys based on the studied parameters. To our best knowledge, the present study is the first to provide a complex analysis of quality parameters of eight unifloral Hungarian honeys.

Honey is a rich source of carbohydrates, while minor compounds such as amino acids and polyphenols contribute to its health-promoting effects. Honey is one of the oldest traditional remedies applied for microbial infections, due to its antibacterial, anti-inflammatory, and antioxidant properties. The aim of this study was to investigate the antibacterial and antibiofilm effects of Hungarian black locust, linden, and sunflower honeys against the most common biofilm-forming respiratory tract pathogens Haemophilus spp., Pseudomonas aeruginosa, and Streptococcus pneumoniae. The unifloral character of all three honey types was confirmed by melissopalynological analysis. The antibacterial activity of each honey sample against each bacterium strain was proven with agar well diffusion assay and thin layer chromatography—direct bioautography. Kinetics and mechanisms of antibacterial action were clarified with time-kill assay and membrane degradation study. The anti-biofilm activity was evidenced using crystal violet assay. In each assay, linden honey was the most effective, followed by sunflower and black locust honey. In addition, each honey sample had greater potential to suppress respiratory tract bacteria, compared to major sugar components. In conclusion, honey in general and linden honey in particular, can have a role in the treatment of respiratory tract infections caused by biofilm-forming bacteria.

Honey is known as an alternative remedy for the treatment of wounds. To evaluate the potential of five Hungarian honey types against wound-associated bacteria, in vitro microbiological assays were conducted on Pseudomonas aeruginosa, Staphylococcus epidermidis and methicillin-resistant Staphylococcus aureus (MRSA). Minimum inhibitory concentration (MIC) was determined with the broth macrodilution method, and biofilm degradation capacity was tested with a crystal violet assay. To understand the underlying mechanisms, the effects of honey treatments were assessed on bacterial membrane integrity and quorum sensing (QS). The highest antibacterial activity, indicated by the lowest MIC values, as well as the highest biofilm inhibition rates and membrane disruption, was displayed by chestnut and linden honeys. The most sensitive bacterium was S. epidermidis. Bacterial membrane degradation took place 40 min after treatment with honey solutions of at least a 40% concentration. Each honey sample exhibited anti-QS activity, which was most pronounced in the case of chestnut honey. It was concluded that the antibacterial, biofilm-inhibiting and anti-QS activities of linden and chestnut honeys were superior to those of acacia, goldenrod and milkweed honeys. In addition to the floral source, the antibacterial effect of honey is influenced by the microbial species treated. The use of honey in wound treatment can be justified by its diverse antibacterial mechanisms.

The irresponsible overuse of antibiotics has increased the occurrence of resistant bacterial strains, which represents one of the biggest patient safety risks today. Due to antibiotic resistance and biofilm formation in bacteria, it is becoming increasingly difficult to suppress the bacterial strains responsible for various chronic infections. Honey was proven to inhibit bacterial growth and biofilm development, offering an alternative solution in the treatment of resistant infections and chronic wounds. Our studies included chestnut honey, valued for its high antibacterial activity, and the bacteria Pseudomonas aeruginosa, methicillin-resistant Staphylococcus aureus, and S. epidermidis, known to form multi-species biofilm communities. Minimum inhibitory concentrations (MIC) of chestnut honey were determined for each bacterial strain. Afterwards, the mixed bacterial biofilms were treated with chestnut honey at different stages of maturity (incubation times: 2, 4, 6, 12, 24 h). The extent of biofilm inhibition was measured with a crystal violet assay and demonstrated by scanning electron microscopy (SEM). As the incubation time increased and the biofilm became more mature, inhibition rates decreased gradually. The most sensitive biofilm was the combination MRSA-S. epidermidis, with a 93.5% inhibition rate after 2 h of incubation. Our results revealed that chestnut honey is suitable for suppressing the initial and moderately mature stages of mixed biofilms.

The eradication of bacterial biofilms remains a persistent challenge in medicine, particularly because an increasing number of biofilms exhibit resistance to conventional antibiotics. This underscores the importance of searching for novel compounds that present antibacterial and biofilm inhibition activity. Various types of honey and essential oil were proven to be effective against a number of biofilm-forming bacterial strains. The current study demonstrated the effectiveness of the relatively unexplored fennel honey (FH), fennel essential oil (FEO), and their combination against biofilm-forming bacterial strains Pseudomonas aeruginosa, methicillin-resistant Staphylococcus aureus, and Escherichia coli, with a series of in vitro experiments. The authenticity of FH and FEO was checked with light microscopy and gas chromatography-mass spectrometry, respectively. Minimum inhibitory concentrations were determined using the microdilution method, and antibiofilm activity was assessed with crystal violet assay. Structural changes in bacterial cells and biofilms, induced by the treatments, were monitored with scanning electron microscopy. FEO and FH inhibited the biofilm formation of each bacterial strain, with FEO being more effective compared to FH. Their combination was the most effective, with inhibitory rates ranging between 87 and 92%, depending on the bacterial strain. The most sensitive bacterium was E. coli, while P. aeruginosa was the most resistant. These results provide justification for the combined use of honey and essential oil to suppress bacterial biofilms and can serve as a starting point to develop an effective surface disinfectant with natural ingredients.

Background/Objectives: Bacterial resistance to antibiotics is a major problem in healthcare, complicated by the ability of bacteria to form biofilms. Complementary therapy for infectious diseases can rely on natural substances with antibacterial activity, e.g., essential oils and honeys. The aim of the study was to investigate the effects of linden and chestnut honeys, lavender essential oil, and their combinations against the multidrug-resistant otitis media pathogens Haemophilus influenzae, H. parainfluenzae, Moraxella catarrhalis, Pseudomonas aeruginosa, and Streptococcus pneumoniae. The efficacy of these natural substances was compared with each other and antibiotics used in clinical practice. Methods: Microscopic pollen analysis and physicochemical traits were used to confirm the botanical origin of honey samples. The antibiotic sensitivity of bacteria was tested with a disk diffusion assay. Minimum inhibitory concentrations were determined using a microdilution assay. A 24 h immature biofilm eradication test was performed with a crystal violet assay. The efficacy of combinations was tested with a checkerboard titration method. The DNA release of damaged bacterial cells was measured using a membrane degradation assay. Results: Lavender essential oil displayed more potent antibacterial activity compared to the honey samples. However, honey–essential oil combinations showed higher inhibition rates for biofilm eradication, with P. aeruginosa being the most resistant bacterium. The combined use of chestnut honey and lavender oil resulted in a higher degree of membrane degradation in a shorter time, and their synergistic effect was proven with checkerboard titration. Conclusions: The combination of linden or chestnut honey with lavender essential oil was shown to be effective in the eradication of a 24 h immature biofilm formed by H. parainfluenzae, M. catarrhalis, and S. pneumoniae.