Edible films can be used as carriers for antimicrobial compounds to assure food safety and quality; in addition, pathogenesis of food bacteria is related to a cell to cell communication mechanism called quorum sensing (QS). Oregano essential oil (OEO) has proved to be useful as food antimicrobial; however, its food applications can be compromised by the volatile character of its active constituents. Therefore, formulation of edible films containing OEO can be an alternative to improve its food usages. QS inhibitory activity of OEO and pectin-OEO films was evaluated using Chromobacterium violaceum as bacterial model. Additionally, antibacterial activity was tested against Escherichia coli O157:H7, Salmonella Choleraesuis, Staphylococcus aureus, and Listeria monocytogenes. OEO was effective to inhibit bacterial growth at MIC of 0.24 mg/mL for all tested bacteria and MBC of 0.24, 0.24, 0.48, and 0.24 mg/mL against E. coli O157:H7, S. Choleraesuis, S. aureus, and L. monocytogenes, respectively. Pectin-films incorporated with 36.1 and 25.9 mg/mL of OEO showed inhibition diameters of 16.3 and 15.2 mm for E. coli O157:H7; 18.1 and 24.2 mm for S. Choleraesuis; 20.8 and 20.3 mm for S. aureus; 21.3 and 19.3 mm for L. monocytogenes, respectively. Pectin-OEO film (15.7 mg/mL) was effective against E. coli O157:H7 (9.3 mm), S. aureus (9.7 mm), and L. monocytogenes (9.2 mm), but not for S. Choleraesuis. All concentrations of OEO (0.0156, 0.0312, 0.0625 and 0.125 mg/mL) and pectin-OEO films (15.7, 25.9 and 36.1 mg/mL) showed a significant anti-QS activity expressed as inhibition of violacein production by C. violaceum. Additionally, the application of pectin-OEO films was effective reducing total coliforms, yeast, and molds of shrimp and cucumber slices stored at 4°C during 15 d. These results demonstrated the potential of pectin films enriched with OEO as food related microorganisms and QS inhibitors.
The main goal of this study was to evaluate the inhibition of Pseudomonas aeruginosa virulence factors and Quorum Sensing during exposure to carvacrol. P. aeruginosa (ATCC 10154) was exposed to carvacrol determining changes in biofilm development, motility, acyl-homoserine lactones (AHL) synthesis and relative expression of lasI/lasR. Docking analysis was used to determinate interactions between carvacrol with LasI and LasR proteins. P. aeruginosa produced 60% lower AHLs when exposed to carvacrol (1.9 mM) compared to control, without affecting cellular viability, indicating a reduction on the LasI synthase activity. AHL-C12, C6, and C4 were detected and related to biofilm development, motility, and pyocyanin production, respectively. The presence of carvacrol reduced the expression of lasR, without affecting lasI gen. Moreover, computational docking showed interactions of carvacrol with amino acids in the active site pocket of LasI (−5.6 kcal mol −1 ) and within the binding pocket of LasR (−6.7 kcal mol −1 ) of P. aeruginosa. These results demonstrated that virulence of P. aeruginosa was reduced by carvacrol, by inhibiting LasI activity with the concomitant reduction on the expression of lasR, biofilm and swarming motility. This study provides relevant information about the effect of carvacrol against quorum sensing to inhibit virulence factors of P. aeruginosa at enzymatic and gene levels. These findings can contribute to the development of natural anti-QS products, which can affect pathogenesis.
The objective of the present study is to evaluate the effect of individual and combined coatings of chitosan (0.008 g·mL−1) and carnauba wax (0.1 g·mL−1) with oregano essential oil (OEO, 0.08 g·mL−1) to reduce dehydration and microbial decay of fresh cucumbers stored at 10 °C. Chitosan-OEO-wax films showed the lowest water vapor transmission rate (0.141 g·m−2·h−1), compared to single chitosan films (0.257 g·m−2·h−1). While chitosan-OEO films completely inhibited the in vitro growth of Alternaria alternata and reduced the growth of Salmonella Typhimurium, Escherichia coli O157:H7, mesophilic bacteria, and fungi isolated from decayed cucumbers. Besides, the infrared analysis of chitosan-OEO-wax films showed shifts in O–H and N–H absorption bands, indicating possible hydrogen bonding between the components. Wax and wax-OEO were the most effective coatings to prevent weight loss in cucumbers during 15 days of storage at 10 °C, while the most effective antimicrobial treatments were chitosan and chitosan-OEO. Therefore, these results showed that carnauba wax and carnauba wax-OEO coatings were the most effective in weight loss, whereas chitosan and chitosan-OEO were the most effective to reduce the microbial load of the treated fresh cucumber.
Aims
To study the individual and combined contribution of catechin, protocatechuic and vanillic acids to inhibit the adhesion of uropathogenic Escherichia coli (UPEC) on the surface of silicone catheters.
Methods and Results
The adhesion of UPEC to silicone catheters during the exposure to nonlethal concentrations of phenolic compounds was measured, as well as changes in motility, presence of fimbriae, extra-cellular polymeric substances, surface charge, hydrophobicity and membrane fluidity. The phenolic combination reduced 26–51% of motility, 1 log CFU per cm2 of adhered bacteria and 20–40% the carbohydrate and protein content in the biofilm matrix. Curli fimbriae, surface charge and cell hydrophobicity were affected to a greater extent by the phenolic combination. In the mixture, vanillic acid was the most effective for reducing bacterial adhesion, extra-polymeric substance production, motility, curli fimbriae and biofilm structure. Notwithstanding, protocatechuic acid caused major changes in the bacterial cell surface properties, whereas catechin affected the cell membrane functionality.
Conclusion
Catechin, protocatechuic and vanillic acids have different bacterial cell targets, explaining the synergistic effect of their combination against uropathogenic E. coli.
Significance and Impact of Study
This study shows the contribution of catechin, protocatechuic and vanillic acids in producing a synergistic mixture against the adhesion of uropathogenic E. coli on silicone catheters. The action of catechin, vanillic and protocatechuic acids included specific contributions of each compound against the E. coli membrane’s integrity, motility, surface properties and production of extracellular polymeric substances. Therefore, the studied mixture of phenolic compounds could be used as an antibiotic alternative to reduce urinary tract infections associated with silicone catheters.
The resistance of Escherichia coli O157:H7 to disinfection is associated with its ability to form biofilms, mainly constituted by glucans produced by glucosyltransferases. Citral and geraniol, terpenes found in the essential oil of Cymbopogon citratus (EO), have proven antibacterial activity against planktonic E. coli; however, no information was found about their efficacy and mode of action against E. coli biofilms. Therefore, the inhibitory effect of C. citratus EO, citral, and geraniol on glucans production and glucosyltransferase activity as anti-biofilm mechanism against E. coli was evaluated. EO, citral, and geraniol inhibited the planktonic growth of E. coli (minimal inhibitory concentration or MIC= 2.2, 1.0, and 3.0 mg/mL, respectively) and the bacterial adhesion (2.0, 2.0, and 4.0 mg/mL, respectively) on stainless steel. All compounds decreased the glucans production; citral and geraniol acted as uncompetitive inhibitors of glucosyltransferase activity (The half maximal inhibitory concentrations or IC50 were 8.5 and 6.5 µM, respectively). The evidence collected by docking analysis indicated that both terpenes could interact with the helix finger of the glucosyltransferase responsible for the polymer production. In conclusion, C. citratus EO, citral, and geraniol inhibited glucosyltransferase activity, glucans production, and the consequent biofilm formation of E. coli O157:H7.
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