Nine odorless laboratory-collected hydro-distilled aqueous extracts (basil, calendula, centrifuged oregano, corn silk, laurel, oregano, rosemary, spearmint, thyme) and one industrial steam-distilled ...oregano hydrolate acquired as by-products of essential oils purification were screened for their in vitro antimicrobial activity against three Salmonella Typhimurium strains (4/74, FS8, FS115) at 4 and 37 °C. Susceptibility to the extracts was mainly plant- and temperature-dependent, though strain dependent effects were also observed. Industrial oregano hydrolate eliminated strains immediately after inoculation, exhibiting the highest antimicrobial potential. Hydro-distilled extracts eliminated/reduced Salmonella levels during incubation at 4 °C. At 37 °C, oregano, centrifuged oregano, thyme, calendula and basil were bactericidal while spearmint, rosemary and corn silk bacteriostatic. A strain-dependent effect was observed for laurel. The individual or combined effect of marinades and edible coatings prepared of industrial hydrolate and hydro-distilled oregano extracts with or without oregano essential oil (OEO) was tested in pork meat at 4 °C inoculated with FS8 strain. Lower in situ activity was observed compared to in vitro assays. Marinades and edible coatings prepared of industrial oregano hydrolate + OEO were the most efficient in inhibiting pathogen. Marination in oregano extract and subsequent coating with either 50% oregano extract + OEO or water + OEO enhanced the performance of oregano extract. In conclusion, by-products of oregano essential oil purification may be promising alternative antimicrobials to pork meat stored under refrigeration when applied in the context of multiple hurdle approach.
The foodborne pathogen L. monocytogenes can be present in food processing environments where it is exposed to various stressors. These antimicrobial factors, which aim to eliminate the pathogen, can ...induce sub-lethal injury to the bacterial cells. In the present study, we investigated the efficacy of different treatments (stresses) relevant to food processing and preservation as well as sanitation methods, in generating sub-lethal injury at 4 °C and 20 °C to two L. monocytogenes strains, ScottA and EGDe. Additionally, we evaluated the survival and extent of L. monocytogenes injury after exposure to commonly used disinfectants (peracetic acid and benzalkonium chloride), following habituation in nutrient-deprived, high-salinity medium. Each stress had a different impact on the survival and injury kinetics of L. monocytogenes. The highest injury levels were caused by peracetic acid which, at 4 °C, generated high populations of injured cells without loss of viability. Other injury-inducing stresses were lactic acid and heating. Long-term habituation in nutrient-limited and high salinity medium (4 °C) and subsequent exposure to disinfectants resulted in higher survival and injury in benzalkonium chloride and increased survival, yet with lower injury levels, in peracetic acid at 20 °C. Taken together, these results highlight the potential food safety risk emerging from the occurrence of injured cells by commonly used food processing methods. Consequently, in order to accurately assess the impact of an antimicrobial method, its potential of inducing sublethal injury needs to be considered along with lethality.
•Different food-processing treatments may differently affect L. monocytogenes injury.•Peracetic acid can induce significant levels of injury compared to other stresses.•Temperature, exposure time and dilutant of the stressor affect the extent of injury.•Stress-induced injury may not be correlated with the lethality of the procedure.•Sanitizers-survival and injury was affected following starvation in high salinity.
The disinfectant peracetic acid (PAA) that is used in the food industry can cause sublethal injury in L. monocytogenes. The effect of preculture temperature on the inactivation and sublethal injury ...of L. monocytogenes cells due to PAA was evaluated by plating on non-selective and selective agar medium supplemented with 5 % (w/v) NaCl. L. monocytogenes cells were precultured at 30 °C, 20 °C or 4 °C, and the former was used as reference temperature. Preculture of cells at 20 °C or 4 °C and subsequent exposure to PAA at the respective growth temperatures caused higher injury compared to cells grown at 30 °C and exposed to PAA 20 °C and PAA 4 °C, respectively. Survival was also affected by the preculture temperature; 20 °C-grown cultures resulted in lower survival at PAA 20 °C. Nevertheless, preculture at 4 °C resulted in a similar number of surviving cells when exposed to PAA 4 °C compared to cells precultured at 30 °C and exposed to PAA at 4 °C. Flow cytometry was subsequently used to quantify outgrowth capacity of stressed and sublethal damaged populations following sorting of single cells in nutrient rich medium (Tryptone soy broth supplemented with yeast extract TSBY). PAA treatment affected the outgrowth of L. monocytogenes at single-cell level resulting in increased outgrowth-times reflecting higher single cell heterogeneity. To conclude, the response of L. monocytogenes when exposed to PAA depended on the preculture conditions, and the highly heterogeneous outgrowth potential of PAA-injured cells may affect their detection accuracy and pose a food safety risk.
•Preculturing temperature affected survival and injury upon peracetic acid treatment.•Decrease of preculturing temperature from 30 °C to 20 °C triggered faster reduction of viability.•Reduction of viability upon PAA treatment at 4 °C was less effective than at 20 °C.•PAA treatment at 20 °C increased variability in outgrowth of single cells.
The disinfectant peracetic acid (PAA) can cause high levels of sublethal injury to Listeria monocytogenes. This study aims to evaluate phenotypic and transcriptional characteristics concerning the ...surface attachment and virulence potential of sublethally injured L. monocytogenes ScottA and EGDe after exposure to 0.75 ppm PAA for 90 min at 4°C and subsequent incubation in tryptic soy broth supplemented with yeast extract (TSBY) at 4°C. The results showed that injured L. monocytogenes cells (99% of the total population) were able to attach (after 2 and 24 h) to stainless steel coupons at 4°C and 20°C.
virulence assays using human intestinal epithelial Caco-2 cells showed that injured L. monocytogenes could invade host cells but could not proliferate intracellularly. The
virulence response was strain dependent; injured ScottA was more invasive than EGDe. Assessment of PAA injury at the transcriptional level showed the upregulation of genes (
and
) involved in flagellum motility and surface attachment. The transcriptional responses of L. monocytogenes EGDe and ScottA were different: only injured ScottA demonstrated upregulation of the virulence genes
and
. Downregulation of the stress-related genes
and
and upregulation of
were observed in injured ScottA. The obtained results indicate that sublethally injured L. monocytogenes cells may retain part of their virulence properties as well as their ability to adhere to food-processing surfaces. Transmission to food products and the introduction of these cells into the food chain are therefore plausible scenarios that are worth taking into consideration in terms of risk assessment.
L. monocytogenes is the causative agent of listeriosis, a serious foodborne illness. Antimicrobial practices such as disinfectants used for the elimination of this pathogen in the food industry can produce a sublethally injured population fraction. Injured cells of this pathogen that may survive antimicrobial treatment may pose a food safety risk. Nevertheless, knowledge regarding how sublethal injury may impact important cellular traits and phenotypic responses of this pathogen is limited. This work suggests that sublethally injured L. monocytogenes cells maintain virulence and surface attachment potential and highlights the importance of the occurrence of sublethally injured cells regarding food safety.
The effect of the degree of toasting of oak chips used in wine ageing on the extraction pattern of polyphenols was investigated using model wine systems. Pattern identification was carried out ...employing a Box–Behnken experimental design and response surface methodology, using resident time (t) and amount of chips (ACₕᵢₚ) as the independent variables. The responses selected to assess the process were the total polyphenol concentration, the antiradical activity and the reducing power of the model wines. Further, a deeper insight into the effects exerted by chip toasting was provided by the examination of the analytical polyphenolic composition of model wines, using liquid chromatography–mass spectrometry. The results obtained demonstrated that even moderate toasting provoked drastic decrease in the native oak wood polyphenols (ellagitannins), as indicated by the Folin–Ciocalteu assay, with a concomitant reduction in both the antiradical activity and reducing power. The evolution pattern for all three independent variables during model wine treatment with chips having different degree of toasting was also substantially affected. Light toasting was shown to generate a series of small molecular weight phenolics, but some of them did not survive more intense (heavy) toasting.
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