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Honey exhibited potential antimicrobial activity against multidrug resistant (MDR) bacteria that continues to be a serious health problem. We reported the in-vitro activity of Saudi Sumra honey against clinical pathogenic bacteria and fungi, antibiofilm, anti-quorum-sensing (QS) and antioxidant activities in relation to its phytochemical composition assessed by gas chromatography-mass spectrometry (GC-MS). Broth dilution method and scavenging activities against 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and <i>β</i>-carotene bleaching assays were performed. The GC-MS analysis of Sumra honey showed that 2,4-dihydroxy-2,5-dimethyl-3(2H)-furan-3-one 1-methylcyclopropanemethanol were the major identified phytoconstituents. Sumra honey showed a minimum inhibitory concentration (MIC) to clinical isolates of <i>Staphylococcus aureus</i> including methicillin-resistant <i>Staphylococcus aureus</i> (MRSA) at 300 mg/mL, <i>Pseudomonas aeruginosa</i> (250 mg/mL), <i>Escherichia coli</i> (350 mg/mL) and <i>Acinetobacter baumannii</i> (250 mg/mL); clinical fungal isolates—<i>Candida auris</i> (600 mg/mL) and <i>Cryptococcus neoformans</i> (>1000 mg/mL); wild type fungal isolates—<i>Candida krusei</i> (>1000 mg/mL) and <i>Candida albicans</i> (700 mg/mL). In addition, Sumra honey demonstrated promising inhibition targeting biofilm formation by 59% for <i>Bacillus subtilis</i>, 48% for <i>S. aureus</i>, 38% for <i>E. coli</i>, and 33.63% for <i>P. aeruginosa</i>. The violacein production in <i>Chromobacterium violaceum</i> was reduced to 68%, whereas pyocyanin production in <i>P. aeruginosa</i> was reduced to 54.86% at ½ MIC. Furthermore, Sumra honey exhibited strong antioxidant activities (DPPH − IC₅₀ = 7.7 mg/mL; ABTS − IC₅₀ = 5.4 mg/mL; <i>β</i>-carotene − IC₅₀ = >20 mg/mL). Overall, obtained data highlighted the promising potential therapeutic use of Sumra honey treating infections caused by MDR bacteria and fungi. Moreover, Sumra honey can be a good candidate as an inhibitor agent for bacterial cellular communication in strains of <i>P. aeruginosa</i> and <i>C. violaceum</i>.