The role of lactic acid bacteria (LAB) in honey as antifungal activity has received little attention and their mechanism of inhibitory of fungi is not fully understood. In this study, LAB were isolated from honey samples from Malaysia, Libya, Saudi Arabia, and Yemen. Twenty-five isolates were confirmed LAB by catalase test and Gram staining, and were screened for antifungal activity. Four LAB showed inhibitory activity against Candida spp. using the dual agar overlay method. And they were identified as Lactobacillus plantarum HS isolated from Al-Seder honey, Lactobacillus curvatus HH isolated from Al-Hanon honey, Pediococcus acidilactici HC isolated from Tualang honey and Pediococcus pentosaceus HM isolated from Al-Maray honey by the 16S rDNA sequence. The growth of Candida glabrata ATCC 2001 was strongly inhibited (>15.0 mm) and (10~15 mm) by the isolates of L. curvatus HH and P. pentosaceus HM, respectively. The antifungal activity of the crude supernatant (cell free supernatant, CFS) was evaluated using well diffusion method. The CFS showed high antifungal activity against Candida spp. especially The CFS of L. curvatus HH was significantly (p < 0.05) inhibited growth of C. glabrata ATCC 2001, C. parapsilosis ATCC 2201, and C. tropicalis ATCC 750 with inhibitory zone 22.0, 15.6, and 14.7 mm, respectively. While CFS of P. pentosaceus HM was significantly (p < 0.05) effective against C. krusei, C. glabrata, and C. albicans with inhibition zone 17.2, 16.0, and 13.3 mm, respectively. The results indicated that LAB isolated from honey produced compounds which can be used to inhibit the growth of the pathogenic Candida species.

:  A total of 32 lactic acid bacteria (LAB) were isolated from 13 honey samples commercially marketed in Malaysia, 6 strains identified as Lactobacillus acidophilus by API CHL50. The isolates had antibacterial activities against multiple antibiotic resistant's Staphylococcus aureus (25 to 32 mm), Staphylococcus epidermis (14 to 22 mm) and Bacillus subtilis (12 to 19 mm) in the agar overlay method after 24 h incubation at 30 °C. The crude supernatant was heat stable at 90 °C and 121 °C for 1 h. Treatment with proteinase K and RNase II maintained the antimicrobial activity of all the supernatants except sample H006‐A and H010‐G. All the supernatants showed antimicrobial activities against target bacteria at pH 3 and pH 5 but not at pH 6 within 72 h incubation at 30 °C. S. aureus was not inhibited by sample H006‐A isolated from Libyan honey and sample H008‐D isolated from Malaysian honey at pH 5, compared to supernatants from other L. acidophilus isolates. The presence of different strains of L. acidophilus in honey obtained from different sources may contribute to the differences in the antimicrobial properties of honey.

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The stingless bee Heterotrigona itama honey is generating interest among consumers due to unique antibacterial properties. Despite the substantial health benefits of this honey, its action mode of inhibiting and killing pathogenic bacteria remains unresolved. The present study aimed to unravel the effect of raw H. itama honey against four clinically important bacteria, Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis and Staphylococcus aureus. Total phenolic and total flavonoid contents, alongside antioxidant and antibacterial activities of honey samples were established to identify the 'best' raw honey. Honey Sy-1 showing the highest amount of phenolics (80.71 mg GAE/100 g) and antioxidant capacity (82.67%) and, the lowest MIC (1.56% (w/v)) and MBC (3.125% (w/v)), was tested against four bacterial cells, wherein any alterations in cellular morphology were observed under scanning electron microscope. The micrographs revealed significant elongation/filamentation in B. subtilis cells, de-flagellation in P. aeruginosa, membrane perturbation and leakage of intracellular contents in E. coli alongside non-dividing septated cells in S. aureus. H. itama honey inhibited bacterial growth by disrupting the cell membrane in conjunction to interacting with a cytosolic target or possibly interrupted septum formation and blocking DNA and protein syntheses. The evidence strongly supported the natural antibacterial properties of the H. itama honey.

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