Transcriptomic approach and membrane fatty acid analysis to study the response mechanisms of Escherichia coli to thyme essential oil, carvacrol, 2-(E)-hexanal and citral exposure (original) (raw)
Related papers
Journal of Agricultural and Food Chemistry, 2006
Major active compounds from essential oils are well-known to possess antimicrobial activity against both pathogen and spoilage microorganisms. The aim of this work was to determine the alteration of the membrane fatty acid profile as an adaptive mechanism of the cells in the presence of a sublethal concentration of antimicrobial compound in response to a stress condition. Methanolic solutions of thymol, carvacrol, limonene, cinnamaldehyde, and eugenol were added into growth media of Escherichia coli O157:H7, Salmonella enterica serovar typhimurium, Pseudomonas fluorescens, Brochothrix thermosphacta, and Staphylococcus aureus strains. Fatty acid extraction and gas chromatographic analysis were performed to assess changes in membrane fatty acid composition. Substantial changes were observed on the long chain unsaturated fatty acids when the E. coli and Salmonella strains grew in the presence of limonene and cinnamaldehyde and carvacrol and eugenol, respectively. All compounds influenced the fatty acid profile of B. thermosphacta, while Pseudomonas and S. aureus strains did not show substantial changes in their fatty acid compositions.
FEMS Microbiology Letters, 2014
The aim of the current study was to determine, via reverse transcription quantitative polymerase chain reaction (RT-qPCR) analysis, the effect of oregano essential oil (Origanum heracleoticum) and carvacrol, its major component, on the expression of virulence-associated genes in enterohaemorrhagic Escherichia coli (EHEC) O157:H7 ATCC strain 35150. Both oregano oil and carvacrol demonstrated their efficacy firstly, by inhibiting the transcription of the ler gene involved in upregulation of the LEE2, LEE3 and LEE4 promoters and of attaching and effacing lesions and secondly by decreasing both Shiga toxin and fliC genes expression. In addition, a decrease in luxS gene transcription involved in quorum sensing was observed. These results were dose dependent and showed a specific effect of O. heracleoticum and carvacrol in downregulating the expression of virulence genes in EHEC O157:H7. These findings suggest that oregano oil and carvacrol have the potential to mitigate the adverse health effects caused by virulence gene expression in EHEC O157:H7, through the use of these substances as natural antibacterial additives in foods or as an alternative to antibiotics.
Food Research International, 2022
Essential oils (EOs), such as thyme (Thy) and cinnamon (Cin) oils, present promising antibacterial properties against foodborne pathogens (e.g., Salmonella enterica serovar Typhimurium). However, the food matrix might result in sublethal EO stress, and little information about direct and/or cross-resistance development after sublethal EO exposure is available. This study revealed that S. Typhimurium under sublethal Thy and Cin (50% minimum inhibitory concentration, MIC 50) treatments exhibited a lower growth rate and an extended lag phase. EO adapted cells showed direct-resistance to subsequent lethal EO treatment, and cross-resistance to thermal (58 • C) and oxidative (hydrogen peroxide, 50 mmol/L) stresses. Metabolomics analysis revealed changes of 47 significant metabolites (variable importance in projection > 1, false discovery rate (FDR) < 0.05), including lipids, oligopeptides, amino acids, nucleotide related compounds, and organic acids. Metabolic pathways, such as aminoacyl-tRNA biosynthesis, were shown to be involved in EO adaptation. Furthermore, a transcriptomics study identified 161 differentially expressed genes (DEGs, fold change > 2, FDR < 0.05) in MIC 50 Thy treated cells, while more DEGs (3 2 4) were screened from the MIC 50 Cin group. The integrated omics analysis allowed us to speculate on the molecular mechanisms. Under harsher Thy stress, S. Typhimurium cells adopted a conservative strategy to survive. By contrast, more radical responses were observed during Cin adaptation. In conclusion, the food industry should be more cautious in the use of EOs because sublethal EO stress might result in the development of resistance.
Membrane Toxicity of Antimicrobial Compounds from Essential Oils
Journal of Agricultural and Food Chemistry, 2007
Natural antimicrobial compounds perform their action mainly against cell membranes. The aim of this work was to evaluate the interaction, meant as a mechanism of action, of essential oil antimicrobial compounds with the microbial cell envelope. The lipid profiles of Escherichia coli O157:H7, Staphylococcus aureus, Salmonella enterica serovar Typhimurium, Pseudomonas fluorescens, and Brochothrix thermosphacta cells treated with thymol, carvacrol, limonene, eugenol, and cinnamaldehyde have been analyzed by gas chromatography. In line with the fatty acids analysis, the treated cells were also observed by scanning electron microscopy (SEM) to evaluate structural alterations. The overall results showed a strong decrease of the unsaturated fatty acids (UFAs) for the treated cells; in particular, the C18:2trans and C18:3cis underwent a notable reduction contributing to the total UFA decreases, while the saturated fatty acid C17:0 raised the highest concentration in cinnamaldehyde-treated cells. SEM images showed that the used antimicrobial compounds quickly exerted their antimicrobial activities, determining structural alterations of the cell envelope.
Journal of Food …, 2005
The major components of oregano and thyme essential oils that had previously been shown to inhibit Escherichia coli O157:H7 were determined by high-performance liquid chromatography with UV detection and liquid chromatographic tandem mass spectrometry. The MICs and MBCs of carvacrol, thymol, p-cymene, and ␥-terpinene against a strain of E. coli O157: H7 phage type 34 isolated from bovine feces were determined by microdilution assay. The constituents were then tested in checkerboard assays to detect possible interactions. Carvacrol and thymol displayed bacteriostatic and bactericidal properties with MICs of 1.2 mmol/liter and were additive in combination. p-Cymene and ␥-terpinene displayed no measurable antibacterial activity up to 50 mmol/liter, and neither influenced the activity of carvacrol or thymol. Growth curves in the presence of nonlethal concentrations of carvacrol with the addition of agar (0.05%, wt/vol) or carrageenan (0.125%, wt/vol) as stabilizer were produced by optical density measurement. The stabilizers agar and carrageenan both significantly improved the effectiveness of carvacrol in broth, possibly because of a delay in the separation of the hydrophobic substrate from the aqueous phase of the medium. When carvacrol was dissolved in ethanol before addition to broth, stabilizers were not needed. Carvacrol and thymol, particularly when used in combination with a stabilizer or in an ethanol solution, may be effective in reducing the number or preventing growth of E. coli O157:H7 in liquid foods.
Current Microbiology, 2020
Abused refrigerated temperatures are described as unacceptable deviations from the optimal temperature, occurring frequently during transportation of food products. Escherichia coli O157:H7 is a serious contaminant of meats and meat products due to its ability to grow at abused temperatures (> 10 °C). The aim of this study was to evaluate the antibacterial activity of Carum copticum essential oil for the control of Escherichia coli O157:H7 using laboratory media and minced beef at severe abused refrigerated temperature (15 °C). A comparative quantitative reverse transcription real-time PCR was used to assess effects of temperature and Carum copticum essential oil at sub-minimum inhibitory concentrations on bacterial growth and Shiga-toxin gene (stx1A and stx2A) expression. Results indicated that Carum copticum essential oil inhibited growth of E. coli O157:H7 in tryptone soy broth (TSB) media at all sub-MIC values until Hour 48. However, bacterial population increased progressively until Hour 72 at essential oil concentration of 0.75% (ml g −1) and reached 8.6 log CFU g −1 in minced beef. The essential oil at concentration of 0.005% (ml g −1) increased stx gene expression at all times, but increased stx gene expression (0.015%) at Hour 24 in TSB media. The expression rate of stx1A in minced beef decreased progressively (10.39 and 7.67 folds for 0.5 and 0.75%, respectively) and expression of stx2A was variable in minced beef during storage. In conclusion, results from this study have shown that effects of Carum copticum essential oil on growth and virulence gene expression are not necessarily correlated and temperature, essential oil concentration, investigated gene type, and bacterial growth environment (in vivo or in vitro) are effective as well.
Applied and Environmental Microbiology, 2011
To enhance food safety and stability, the food industry tends to use natural antimicrobials such as plantderived compounds as an attractive alternative to chemical preservatives. Nonetheless, caution must be exercised in light of the potential for bacterial adaptation to these molecules, a phenomenon previously observed with other antimicrobials. The aim of this study was to characterize the adaptation of Salmonella enterica serovar Typhimurium to sublethal concentrations of four terpenes extracted from aromatic plants: thymol, carvacrol, citral, and eugenol, or combinations thereof. Bacterial adaptation in these conditions was demonstrated by changes in membrane fatty acid composition showing (i) limitation of the cyclization of unsaturated fatty acids to cyclopropane fatty acids when cells entered the stationary phase and (ii) bacterial membrane saturation. Furthermore, we demonstrated an increased cell resistance to the bactericidal activity of two biocides (peracetic acid and didecyl dimethyl ammonium bromide). The implications of membrane modifications in terms of hindering the penetration of antimicrobials through the bacterial membrane are discussed.
Antibiotics, 2021
Biofilm as a cellular conformation confers survival properties to microbial populations and favors microbial resistance. Here, we investigated the antimicrobial, antibiofilm, antimotility, antihemolytic activity, and the interaction with synthetic membranes of 15 essential oils (EOs) on E. coli ATCC 25922 and S. aureus ATCC 29213. Antimicrobial activity of EOs was determined through microdilution method; development of the biofilm was assessed using the crystal violet assay and SEM microscopy. Results indicate that Lippia origanoides thymol–carvacrol II chemotype (LTC II) and Thymus vulgaris (TV) exhibited a significant antibacterial activity, with MIC values of 0.45 and 0.75 mg/mL, respectively. The percentage of biofilm formation inhibition was greater than 70% at subinhibitory concentrations (MIC50) for LTC II EO. The results demonstrate that these two oils had significantly reduced the hemolytic effect of S. aureus by 54% and 32%, respectively, and the mobility capacity by swimm...
Characterization of the action of selected essential oil components on Gram-negative bacteria
Journal of agricultural …, 1998
Carvacrol, (+)-carvone, thymol, and trans-cinnamaldehyde were tested for their inhibitory activity against Escherichia coli O157:H7 and Salmonella typhimurium. In addition, their toxicity to Photobacterium leiognathi was determined, utilizing a bioluminescence assay. Their effects on the cell surface were investigated by measuring the uptake of 1-N-phenylnaphthylamine (NPN), by measuring their sensitization of bacterial suspensions toward detergents and lysozyme, and by analyzing material released from cells upon treatment by these agents. Carvacrol, thymol, and trans-cinnamaldehyde inhibited E. coli and S. typhimurium at 1-3 mM, whereas (+)-carvone was less inhibitory. trans-Cinnamaldehyde was the most inhibitory component toward P. leiognathi. Carvacrol and thymol disintegrated the outer membrane and released outer membrane-associated material from the cells to the external medium; such release by (+)-carvone or trans-cinnamaldehyde was negligible. Of the tested components, carvacrol and thymol decreased the intracellular ATP pool of E. coli and also inreased extracellular ATP, indicating disruptive action on the cytoplasmic membrane.
The Role of Essential Oils against Pathogenic Escherichia coli in Food Products
Microorganisms
Outbreaks related to foodborne diseases are a major concern among health authorities, food industries, and the general public. Escherichia coli (E. coli) is a pathogen associated with causing multiple outbreaks in the last decades linked to several ready to eat products such as meat, fish, dairy products, and vegetables. The ingestion of contaminated food with pathogenic E. coli can cause watery diarrhea, vomiting, and persistent diarrhea as well as more severe effects such as hemorrhagic colitis, end-stage renal disease, and, in some circumstances, hemolytic uremic syndrome. Essential oils (EOs) are natural compounds with broad-spectrum activity against spoilage and pathogenic microorganisms and are also generally recognized as safe (GRAS). Particularly for E. coli, several recent studies have been conducted to study and characterize the effect to inhibit the synthesis of toxins and the proliferation in food systems. Moreover, the strategy used to apply the EO in food plays a cruci...