Synergistic Antibiofilm Effects of Chestnut and Linden Honey with Lavender Essential Oil Against Multidrug-Resistant Otitis Media Pathogens

栗树蜂蜜和椴树蜂蜜与薰衣草精油对多重耐药性中耳炎病原体的协同抗生物膜作用

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Abstract

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.

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