Low concentrations of honey reduce biofilm formation, quorum sensing, and virulence in Escherichia coli O157:H7.
ABSTRACT Bacterial biofilms are associated with persistent infections due to their high resistance to antimicrobial agents. Hence, controlling pathogenic biofilm formation is important in bacteria-related diseases. Honey, at a low concentration of 0.5% (v/v), significantly reduced biofilm formation in enterohemorrhagic Escherichia coli O157:H7 without inhibiting the growth of planktonic cells. Conversely, this concentration did not inhibit commensal E. coli K-12 biofilm formation. Transcriptome analyses showed that honey significantly repressed curli genes (csgBAC), quorum sensing genes (AI-2 importer and indole biosynthesis), and virulence genes (LEE genes). Glucose and fructose in the honeys were found to be key components in reducing biofilm formation by E. coli O157:H7 through the suppression of curli production and AI-2 import. Furthermore, honey, glucose and fructose decreased the colonization of E. coli O157:H7 cells on human HT-29 epithelial cells. These results suggest that low concentrations of honey, such as in honeyed water, can be a practical means for reducing the colonization and virulence of pathogenic E. coli O157:H7.
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ABSTRACT: Microtiter plates with 96 wells have become one of the preferred platforms for biofilm studies mainly because they enable high-throughput assays. In this work, macroscale and microscale methods were used to study the impact of hydrodynamic conditions on the physiology and location of Escherichia coli JM109(DE3) biofilms formed in microtiter plates. Biofilms were formed in shaking and static conditions, and two macroscale parameters were assayed: the total amount of biofilm was measured by the crystal violet assay and the metabolic activity was determined by the resazurin assay. From the macroscale point of view, there were no statistically significant differences between the biofilms formed in static and shaking conditions. However, at a microscale level, the differences between both conditions were revealed using scanning electron microscopy (SEM). It was observed that biofilm morphology and spatial distribution along the wall were different in these conditions. Simulation of the hydrodynamic conditions inside the wells at a microscale was performed by computational fluid dynamics (CFD). These simulations showed that the shear strain rate was unevenly distributed on the walls during shaking conditions and that regions of higher shear strain rate were obtained closer to the air/liquid interface. Additionally, it was shown that wall regions subjected to higher shear strain rates were associated with the formation of biofilms containing cells of smaller size. Conversely, regions with lower shear strain rate were prone to have a more uniform spatial distribution of adhered cells of larger size. The results presented on this work highlight the wealth of information that may be gathered by complementing macroscale approaches with a microscale analysis of the experiments.Journal of Microbiological Methods 10/2013; 95:342-349. · 2.16 Impact Factor
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ABSTRACT: Quorum sensing (QS) is an important regulatory mechanism in biofilm formation and differentiation. Interference with QS can affect biofilm development and antimicrobial susceptibility. This study evaluates the potential of selected phytochemical products to inhibit QS. Three isothiocyanates (allylisothiocyanate - AITC, benzylisothiocyanate - BITC and 2-phenylethylisothiocyanate - PEITC) and six phenolic products (gallic acid - GA, ferulic acid - FA, caffeic acid - CA, phloridzin - PHL, (-) epicatechin - EPI and oleuropein glucoside - OG) were tested. A disc diffusion assay based on pigment inhibition in Chromobacterium violaceum CV12472 was performed. In addition, the mechanisms of QS inhibition (QSI) based on the modulation of N-acyl homoserine lactone (AHLs) activity and synthesis by the phytochemicals were investigated. The cytotoxicity of each product was tested on a cell line of mouse lung fibroblasts. AITC, BITC and PEITC demonstrated a capacity for QSI by modulation of AHL activity and synthesis, interfering the with QS systems of C. violaceum CviI/CviR homologs of LuxI/LuxR systems. The cytotoxic assays demonstrated low effects on the metabolic viability of the fibroblast cell line only for FA, PHL and EPI.Biofouling 12/2013; · 3.40 Impact Factor
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ABSTRACT: Resistance to antibiotics continues to rise and few new therapies are on the horizon. Honey has good antibacterial activity against numerous microorganisms of many different genera and no honey-resistant phenotypes have yet emerged. The mechanisms of antimicrobial activity are just beginning to be understood; however, it is apparent that these are diverse and often specific for certain groups or even species of bacteria. Manuka honey has been most thoroughly characterized and is commercially available as a topical medical treatment for wound infections. Furthermore, since most data are available for this honey, there is a considerable focus on it in this review. It is becoming evident that honeys are more than just bactericidal, as they impact on biofilm formation, quorum sensing and the expression of virulence factors. With this in mind, honey represents an attractive antimicrobial treatment that might have the potential to be used alongside current therapies as a prophylactic or to treat wound infection with multidrug-resistant bacteria in future.Future Microbiology 11/2013; 8:1419-29. · 4.02 Impact Factor