Is skipping the definition of primary and secondary models possible? Prediction of Escherichia coli O157 growth by machine learning.

To predict bacterial population behavior in food, statistical models with specific function form have been applied in the field of predictive microbiology. Modelers need to consider the linear or non-linear relationship between the response and explanatory variables in the statistical modeling approach. In the present study, we focused on machine learning methods to skip definition of primary and secondary structure model. Support vector regression, extremely randomized trees regression, and Gaussian process regression were used to predict population growth of Escherichia coli O157 at 15 and 25 °C without defining the primary and secondary models. Furthermore, the support vector regression model was applied to predict small population of bacteria cells with probability theory. The model performance of the machine learning models were nearly equal to that of the current statistical models. Machine learning models have a potential for predicting bacterial population behavior.

Characterization and Food Application of the Novel Lytic Phage BECP10: Specifically Recognizes the O-polysaccharide of Escherichia coli O157:H7.

Escherichia coli O157:H7 is a global concern that causes serious diseases, such as hemolytic uremic syndrome and bloody diarrhea. To control E. coli O157:H7 in food, a novel siphophage, BECP10, that targets the O157 serotype was isolated and characterized. Unlike other E. coli phages, BECP10 can only infect E. coli O157 strains, and thus, did not infect other strains. The 48 kbp genome of BECP10 contained 76 open reading frames (ORFs), including 33 putative functional ORFs. The phage did not contain lysogeny-related modules or toxin-associated genes, suggesting that the phage might be strictly lytic. The tail spike protein (TSP) sequence had very low homology with the reported T1-like phages, indicating that TSP might be related to this unique host spectrum. The specific O-antigen residue of E. coli O157:H7 may be a key factor for phage infection by adsorption and receptor identification. The phage exhibited strong antibacterial activity against E. coli O157:H7 over a broad pH range and showed little development of phage-insensitive mutants. The phage sustained viability on the burger patties and reduced E. coli O157:H7 to a non-detectable level without the emergence of resistant cells at low temperatures for five days. Therefore, phage BECP10 might be a good biocontrol agent for E. coli O157:H7-contaminated food matrices.

Salt, glucose, glycine, and sucrose protect Escherichia coli O157:H7 against acid treatment in laboratory media.

This study investigated the effects of combinations of acetic or malic acid and various solutes (salt, glucose, glycine, or sucrose) on the survival of Escherichia coli O157:H7 in laboratory broth. Additionally, the effectiveness of combining organic acids and various concentrations of salt (0-18%) or sucrose (0-100%) with different water activity values against E. coli O157:H7 were evaluated. For treatment of 1% malic acid, the addition of 3% salt showed synergistic effect. Whereas, when 3% salt, glucose, glycine, or sucrose was added to 1% acetic acid, the solutes antagonized the action of the acid against E. coli O157:H7. Acetic, lactic, or propionic acid combined with salt at either 7 or 9% or sucrose at 60, 80, or 100% resulted in the highest resistance of E. coli O157:H7. From a result of evaluating the membrane fatty acid (MFA) composition of cells, salt or sucrose significantly increased levels of saturated fatty acids (SFAs) or SFAs and cyclopropane fatty acids, respectively. From the results of this study, the addition of solutes and organic compounds may increase the tolerance of E. coli O157:H7 to acetic, lactic, and propionic acid treatments and that the salt or sucrose significantly affects cell MFA composition.

Peroxyacetic acid and chlorine dioxide unlike chlorine induce viable but non-culturable (VBNC) stage of Listeria monocytogenes and Escherichia coli O157:H7 in wash water.

The elaboration of guidelines for the industry to establish minimum concentration to prevent cross-contamination during washing practices based on operational limits is the core of the recommended criteria for the use of sanitizers. Several studies have evidenced that sanitizers reduced the levels of foodborne pathogens. However, they might lead to the progress into a viable but non-culturable (VBNC) state of the cells. This evidence has raised concerns regarding the effectiveness of the recommended washing practices for the inactivation of microbial cells present in the process wash water (PWW). The present study evaluated if the most commonly used sanitizers, including sodium hypochlorite (chlorine), peroxyacetic acid (PAA) and chlorine dioxide (ClO2) at established operational limits induced the VBNC stage of Listeria monocytogenes and Escherichia coli O157:H7. Prevention of cross-contamination was examined in four different types of PWW from washing shredded lettuce and cabbage, diced onions, and baby spinach under simulated commercial conditions of high organic matter and 1 min contact time. The results obtained for chlorine showed that recommended operational limits (20-25 mg/L free chlorine) were effective in inactivating L. monocytogenes and E. coli O157:H7 in the different PWWs. However, the operational limits established for PAA (80 mg/L) and ClO2 (3 mg/L) reduced the levels of culturable pathogenic bacteria but induced the VBNC state of the remaining cells. Consequently, the operational limits for chlorine are satisfactory to inactivate foodborne pathogens present in PWW and prevent cross-contamination but higher concentrations or longer contact times should be needed for PAA and ClO2 to reduce the likelihood of the induction of VBNC bacteria cells, as it represents a hazard.

Lactic acid bacteria starter in combination with sodium chloride controls pathogenic Escherichia coli (EPEC, ETEC, and EHEC) in kimchi.

Kimchi is one of the primary sources of high sodium content in the Korean diet. Low-sodium kimchi is commercially manufactured to minimize the health effects of high salt. We investigated the influence of lactic acid bacteria (LAB) as starter culture in combination with 1% or 2.5% salt on the survival of pathogenic Escherichia coli and physicochemical properties of kimchi during fermentation at 10 °C and 25 °C. Among ten strains of LAB isolated from kimchi, Leuconostoc mesenteroides (KCTC 13374) and Lactobacillus plantarum (KCTC 33133) exhibited antimicrobial activities against pathogenic E. coli (EPEC, ETEC, and E. coli O157:H7) and strong tolerance to low pH (2 and 3) and 0.3% bile salts. Thus, L. mesenteroides and L. plantarum were used as starter cultures for kimchi that contained 1% and 2.5% salt. All pathogenic E. coli strains survived in kimchi regardless of starter cultures or salt concentration for over 15 days at 10 °C, but they died off within 4 days at 25 °C. Survival of pathogenic E. coli was better in naturally fermented kimchi (titratable acidity:0.65%) than kimchi fermented with starter cultures (titratable acidity:1.0%). At 10 °C, the average delta value of E. coli O157:H7 (16.15 d) was smaller than those of EPEC (20.76 d) and ETEC (20.20 d) in naturally fermented kimchi. Overall, survival ability of E. coli O157:H7 was lower than EPEC and ETEC, although differences were not significant. Reduced salt concentration from 2.5% to 1% in kimchi did not affect the growth of LAB and the fermentation period. Pathogenic E. coli died at a faster rate in kimchi fermented with starter cultures and 1% salt than in naturally fermented kimchi with 2.5% salt.

The role of l-arabinose metabolism for Escherichia coli O157:H7 in edible plants.

Arabinose is a major plant aldopentose in the form of arabinans complexed in cell wall polysaccharides or glycoproteins (AGP), but comparatively rare as a monosaccharide. l-arabinose is an important bacterial metabolite, accessed by pectolytic micro-organisms such as Pectobacterium atrosepticum via pectin and hemicellulose degrading enzymes. However, not all plant-associated microbes encode cell-wall-degrading enzymes, yet can metabolize l-arabinose, raising questions about their use of and access to the glycan in plants. Therefore, we examined l-arabinose metabolism in the food-borne pathogen Escherichia coli O157:H7 (isolate Sakai) during its colonization of plants. l-arabinose metabolism (araBA) and transport (araF) genes were activated at 18 °C in vitro by l-arabinose and expressed over prolonged periods in planta. Although deletion of araBAD did not impact the colonization ability of E. coli O157:H7 (Sakai) on spinach and lettuce plants (both associated with STEC outbreaks), araA was induced on exposure to spinach cell-wall polysaccharides. Furthermore, debranched and arabinan oligosaccharides induced ara metabolism gene expression in vitro, and stimulated modest proliferation, while immobilized pectin did not. Thus, E. coli O157:H7 (Sakai) can utilize pectin/AGP-derived l-arabinose as a metabolite. Furthermore, it differs fundamentally in ara gene organization, transport and regulation from the related pectinolytic species P. atrosepticum, reflective of distinct plant-associated lifestyles.

Inactivation of Escherichia coli O157:H7 on radish and cabbage seeds by combined treatments with gaseous chlorine dioxide and heat at high relative humidity.

This study was done to develop a method to inactivate Escherichia coli O157:H7 on radish and cabbage seeds using simultaneous treatments with gaseous chlorine dioxide (ClO2) and heat at high relative humidity (RH) without decreasing seeds' viability. Gaseous ClO2 was spontaneously vaporized from a solution containing hydrochloric acid (HCl, 1 N) and sodium chlorite (NaClO2, 100,000 ppm). Using a sealed container (1.8 L), an equation (y = 5687×, R2 = 0.9948) based on the amount of gaseous ClO2 generated from HCl-NaClO2 solution at 60 °C and 85% RH was developed. When radish or cabbage seeds were exposed to gaseous ClO2 at concentrations up to 3,000 ppm for 120 min, germination rates did not significantly decrease (P > 0.05). When seeds inoculated with E. coli O157:H7 were treated with 2,000 or 3,000 ppm of gaseous ClO2 in an atmosphere with 85% RH at 60 °C, populations (6.8-6.9 log CFU/g) on both types of seeds were decreased to below the detection limit for enrichment (-0.5 log CFU/g) within 90 min. This study provides useful information for developing a decontamination method to control E. coli O157:H7 and perhaps other foodborne pathogens on plant seeds by simultaneous treatment with gaseous ClO2 and heat at high RH.

Growth temperature influences the resistance of Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium on lettuce to X-ray irradiation.

This study aimed to investigate the effect of different growth temperatures on the resistance of Escherichia coli O157:H7 and Salmonella Typhimurium to low-energy X-ray irradiation. Irradiation of contaminated phosphate-buffered saline with 0.6 kGy X-ray decreased the counts of E. coli O157:H7 cultured at 37 °C to below the detection limit (<1.0 colony-forming unit (CFU)/mL) and those of E. coli O157:H7 cultured at 25 and 15 °C by 4.82 and 4.45 log CFU/mL, respectively. The viable counts of S. Typhimurium cultured at 37, 25, and 15 °C in phosphate-buffered saline decreased by 3.56, 3.08, and 2.75 log CFU/mL, respectively, after irradiation with 0.6 kGy X-ray. Irradiation of contaminated lettuce with 0.4 kGy decreased the counts of E. coli O157:H7 cultured at 37, 25, and 15 °C by 3.97, 3.45, and 3.10 log CFU/cm2, respectively, and those of S. Typhimurium by 4.41, 3.84, and 3.40 log CFU/cm2, respectively. Growth temperature influenced pathogen resistance to X-ray irradiation by modulating cellular membrane and DNA integrity, intracellular enzyme activity, and efflux pump function. The results of this study suggest that the stress resistance status of pathogenic bacteria cultured at different growth temperatures should be considered for the application of X-ray irradiation for fresh produce sterilization.

Evaluating the fate of Escherichia coli O157:H7 and Salmonella spp. on cucumbers.

The occurrence of various foodborne disease outbreaks linked to the consumption of cucumbers worldwide in the last years raised concerns regarding the survival ability of foodborne pathogens on this food matrix. This work aimed at evaluating and quantifying the survival of Escherichia coli O157:H7 and Salmonella spp. on cucumber surfaces. Cucumbers were inoculated with a 5-strain cocktail of each microorganism and kept at 25 °C. The survival ability of two green fluorescent protein (GFP) labelled Salmonella strains inoculated individually on cucumbers was also evaluated. The inoculated areas were swabbed at different time intervals (maximum of 72 h) and cells were enumerated by plate count method (log CFU/cm2). The population of both pathogens decreased significantly on cucumber surfaces over time. E. coli O157:H7 could only be recovered up to 8 h while Salmonella spp. could be detected up to 24 h. The GFP-labelled Salmonella strains showed similar behaviour on cucumbers compared to the evaluated Salmonella cocktail. Survival kinetic parameters were estimated by fitting the Weibull model to the survival data. The data obtained in this study indicate that despite of the rapid decrease on concentrations of both pathogens evaluated on cucumbers surfaces, strategies to avoid their contamination during the supply chain as well as proper cleaning and disinfection protocols must be put forward to mitigate both E. coli O57:H7 and Salmonella on cucumbers and therefore, to decrease the exposure of consumers to microbial hazards and to avoid cross-contamination events during distribution, retail and in domestic environments.

Lactic acid bacteria biofilms and their ability to mitigate Escherichia coli O157:H7 surface colonization.

Lactic acid bacteria (LAB) exert antagonistic activities against diverse microorganisms, including pathogens. In this work, we aimed to investigate the ability of LAB strains isolated from food to produce biofilms and to inhibit growth and surface colonization of Enterohaemorrhagic Escherichia coli (EHEC) O157:H7 at 10°C. The ability of 100 isolated LAB to inhibit EHEC O157:H7 NCTC12900 growth was evaluated in agar diffusion assays. Thirty-seven LAB strains showed strong growth inhibitory effect on EHEC. The highest inhibitory activities corresponded to LAB strains belonging to Lactiplantibacillus plantarum, Pediococcus acidilactici and Pediococcus pentosaceus species. Eighteen out of the 37 strains that showed growth inhibitory effects on EHEC also had the ability to form biofilms on polystyrene surfaces at 10°C and 30°C. Pre-established biofilms on polystyrene of four of these LAB strains were able to reduce significantly surface colonization by EHEC at low temperature (10°C). Among these four strains, Lact. plantarum CRL 1075 not only inhibited EHEC but also was able to grow in the presence of the enteric pathogen. Therefore, this strain proved to be a good candidate for further technological studies oriented to its application in food-processing environments to mitigate undesirable surface contaminations of E. coli.

Antibacterial Activity of Caffeic Acid Combined with UV-A Light against Escherichia coli O157:H7, Salmonella enterica Serovar Typhimurium, and Listeria monocytogenes.

The aim of this study was to evaluate the antibacterial activity of caffeic acid (CA), which is a natural polyphenol, combined with UV-A light against the representative foodborne bacteria Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium, and Listeria monocytogenes. Data regarding the inactivation of these bacteria and its dependence on CA concentration, light wavelength, and light dose were obtained. E. coli O157:H7 and Salmonella Typhimurium were reduced to the detection limit when treated with 3 mM CA and UV-A for 3 J/cm2 and 4 J/cm2, respectively, and 5 J/cm2 treatment induced 3.10 log reduction in L. monocytogenes. To investigate the mechanism for inactivation of Salmonella Typhimurium and L. monocytogenes, measurement of polyphenol uptake, membrane damage assessment, enzymatic activity assay, and transmission electron microscopy (TEM) were conducted. It was revealed that CA was significantly (P < 0.05) absorbed by bacterial cells, and UV-A light allowed a higher uptake of CA for both pathogens. Additionally, CA plus UV-A treatment induced significant (P < 0.05) cell membrane damage. In the enzymatic activity assay, the activities of both pathogens were reduced by CA, and a greater reduction occurred by use of CA plus UV-A. Moreover, transmission electron microscopy (TEM) images indicated that CA plus UV-A treatment notably destroyed the intercellular structure. In addition, antibacterial activity was also observed in commercial apple juice, which showed results similar to those obtained from phosphate-buffered saline (PBS), resulting in a significant (P < 0.05) reduction for all three pathogens without any changes in color parameters (L*, a*, and b*), total phenolic compounds, and DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging activity. IMPORTANCE Photodynamic inactivation (PDI), which involves photoactivation of a photosensitizer (PS), is an emerging field of study, as it effectively reduces various kinds of microorganisms. Although there are several PSs that have been used for PDI, there is a need to find naturally occurring PSs for safer application in the food industry. Caffeic acid, a natural polyphenol found in most fruits and vegetables, has recently been studied for its potential to act as a novel photosensitizer. However, no studies have been conducted regarding its antibacterial activity depending on treatment conditions and its antibacterial mechanism. In this study, we closely examined the effectiveness of caffeic acid in combination with UV-A light for inactivating representative foodborne bacteria in liquid medium. Therefore, the results of this research are expected to be utilized as basic data for future application of caffeic acid in PDI, especially when controlling pathogens in liquid food processing.

Assessment of butylparaben (4-hydroxybenzoic acid butyl ester)-assisted heat treatment against Escherichia coli O157:H7 and Salmonella enterica serotype Typhimurium in meat and bone meal.

Heat-resistant foodborne pathogens have been a concern in low-moisture foods and ingredients (LMFs). Due to low thermal conductivity of low moisture materials, thermal treatment is not efficient and may cause nutritional loss. This study investigated the enhancement of thermal treatment of meat and bone meal (MBM) at low water activity (aw ) by inclusion of butylparaben (BP) as a model antimicrobial compound. Stationary phase Escherichia coli O157:H7 (Shiga toxin-negative) or Salmonella enterica serotype Typhimurium was inoculated into MBM containing 0-2000 ppm BP and incubated at 55 or 60°C for up to 5 hr. A biphasic inactivation pattern was observed for both pathogens, indicating existence of potentially thermal resistant subpopulations. Addition of 1000 ppm BP to MBM (aw  = 0.4) significantly lowered the D-value at 55°C for E. coli O157:H7 (2.6 ± 0.5 hr) compared to thermal treatment alone (5.1 ± 0.6 h) during the treatment after the first 1 hr (p < 0.05), indicating that addition of BP accelerated the inactivation of thermal-resistant subpopulation of E. coli O157:H7 in MBM. Interestingly, similar enhancement in thermal inactivation upon addition of BP was not observed in either the sensitive or resistant subpopulation of S. Typhimurium at aw of 0.4 or 0.7, which is likely caused by the higher thermal resistance developed by S. Typhimurium within a low aw environment (aw  < 0.85). These results suggest that addition of certain antimicrobial compounds can improve the thermal processing efficiency in LMFs, while their efficiency against different pathogens may vary. PRACTICAL APPLICATION: Addition of appropriate food-grade compounds may help to improve thermal treatment efficiency in low moisture foods with varied efficiency against different pathogens. This approach has the potential to reduce the required heat treatment intensity while minimizing food safety risk.

High pressure homogenization inactivation of Escherichia coli and Staphylococcus aureus in phosphate buffered saline, milk and apple juice.

High pressure homogenization (HPH) offers new opportunities for food pasteurization/sterilization. Escherichia coli and Staphylococcus aureus suspended in phosphate buffered saline (PBS) buffer, milk and apple juice at initial concentration of ~106 log10 CFU per ml were subjected to HPH treatments up to 200 MPa with inlet temperatures at 4-40°C. After HPH at 200 MPa with the inlet temperature at 40°C, the count of E. coli suspended in PBS, milk and apple juice reduced by 3·42, 3·67 and 3·19 log10 CFU per ml respectively while the count of S. aureus decreased by 2·21, 1·02 and 2·33 log10 CFU per ml respectively suggesting that S. aureus was more resistant. The inactivation data were well fitted by the polynomial equation. Milk could provide a protective effect for S. aureus against HPH. After HPH at 200 MPa with the inlet temperature at 20°C, the cell structure of E. coli was destroyed, while no obvious damages were found for S. aureus.

Models for factors influencing pathogen survival in low water activity foods from literature data are highly significant but show large unexplained variance.

Factors that control pathogen survival in low water activity foods are not well understood and vary greatly from food to food. A literature search was performed to locate data on the survival of foodborne pathogens in low-water activity (<0.70) foods held at temperatures <37 °C. Data were extracted from 67 publications and simple linear regression models were fit to each data set to estimate log linear rates of change. Multiple linear stepwise regression models for factors influencing survival rate were developed. Subset regression modeling gave relatively low adjusted R2 values of 0.33, 0.37, and 0.48 for Salmonella, E. coli and L. monocytogenes respectively, but all subset models were highly significant (p < 1.0e-9). Subset regression models showed that Salmonella survival was significantly (p < 0.05) influenced by temperature, serovar and strain type, water activity, inoculum preparation method, and inoculation method. E. coli survival was significantly influenced by temperature, water activity, and inoculum preparation. L. monocytogenes survival was significantly influenced by temperature, serovar and strain type, and inoculum preparation method. While many factors were highly significant (p < 0.001), the high degrees of variability show that there is still much to learn about the factors which govern pathogen survival in low water activity foods.

High concentration of sodium chloride could induce the viable and culturable states of Escherichia coli O157:H7 and Salmonella enterica serovar Enteritidis.

In the present study, Escherichia coli O157:H7 and Salmonella enterica serovar Enteritidis were transferred into Luria-Bertani medium without NaCl (LBWS) and adjusted to various pHs (4, 5, 6 and 7) with lactic acid containing 0·75, 5, 10 and 30% NaCl, and stored at 25°C until the bacterial populations reached below detectable levels on tryptic soy agar (TSA). Although E. coli O157:H7 and S. Enteritidis did not grow on TSA when incubated in LBWS with 30% NaCl for 35 and 7 days, more than 60 and 70% of the bacterial cells were shown to be viable via fluorescent staining with SYTO9 and propidium iodide (PI), respectively, suggesting that a number of cells could be induced into the viable but nonculturable (VBNC) state. These bacteria that were induced into a VBNC state were transferred to a newly prepared tryptic soy broth (TSB) and then incubated at 37°C for several days. After more than 7 days, E. coli O157:H7 and S. Enteritidis regained their culturability. We, therefore, suggest that E. coli O157:H7 and S. Enteritidis entered the VBNC state under the adverse condition of higher salt concentrations and were revived when these conditions were reversed.

Polycaprolactone film functionalized with bacteriophage T4 promotes antibacterial activity of food packaging toward Escherichia coli.

Bacteriophages (phages) have been extensively utilized as antibacterial agents in the food industry because of their host-specificity. However, their application in polymer films has been limited because of the lack of a strong attachment method for phage to the surface. We developed an antibacterial film by covalently immobilizing Escherichia coli (E. coli)-specific phage T4 on a polycaprolactone (PCL) film. The chemical bond formation was confirmed by XPS analysis, and the covalent attachment of phage T4 effectively inhibited E. coli growth even after external stimulation of the film by sonication. When applied as a packaging film for raw beef inoculated with E. coli O157:H7, the chemically functionalized PCL film showed approximately 30-fold higher bacterial inhibitory effects than the film with physically adsorbed phage T4. These results indicate the promising application potential of chemically functionalized PCL film with phage T4 as an antibacterial food packaging material against the foodborne pathogen E. coli.

Application of plasma activated water for decontamination of alfalfa and mung bean seeds.

Microbial contamination of fresh produce is a major public health concern, with the number of associated disease outbreaks increasing in recent years. The consumption of sprouted beans and seeds is of particular concern, as these foodstuffs are generally consumed raw, and are produced in conditions favourable for the growth of zoonotic pathogens, if present in seeds prior to sprouting or in irrigation water. This work aimed to evaluate the activity of plasma activated water (PAW) as a disinfecting agent for alfalfa (Medicago sativa) and mung bean (Vigna radiata) seeds, during seed soaking. Each seed type was inoculated with Escherichia coli O157, E. coli O104, Listeria monocytogenes or Salmonella Montevideo, and treated with PAW for different times. A combination of PAW and ultrasound treatment was also evaluated. The germination and growth rate of both seeds were assessed after PAW treatments. PAW was demonstrated to have disinfecting ability on sprouted seeds, with reductions of up to Log10 1.67 cfu/g in alfalfa seeds inoculated with E. coli O104, and a reduction of Log10 1.76 cfu/g for mung bean seeds inoculated with E. coli O157 observed. The germination and growth rate of alfalfa and mung bean sprouts were not affected by the PAW treatments. The combination of a PAW treatment and ultrasound resulted in increased antimicrobial activity, with a reduction of Log10 3.48 cfu/g of S. Montevideo in mung bean seeds observed. These results demonstrate the potential for PAW to be used for the inactivation of pathogenic microorganisms which may be present on sprouted seeds and beans, thereby providing greater assurance of produce safety.

Value-added switchgrass extractives for reduction of Escherichia coli O157:H7 and Salmonella Typhimurium populations on Formica coupons.

Recurring outbreaks linked to Escherichia coli O157:H7-contaminated lettuce and Salmonella enterica-contaminated sprouts highlight the need for improved food safety measures. The aim of this study was to determine the ability of a bio-based antimicrobial extract prepared from switchgrass, a dedicated energy crop, to reduce E. coli O157:H7 and S. Typhimurium populations on Formica coupons, a model food-contact surface. Overnight cultures of ~7 log CFU/mL E. coli O157:H7 and S. Typhimurium, air-dried on Formica coupons were treated with 0.625% NaClO, 70% ethanol, sterile water or different batches of switchgrass extractives (SE1, SE2, and SE3) for up to 30 min. E. coli O157:H7 was reduced by 4.43 log CFU/mL after 1 min by SE3, and to non-detectable levels after 1 min by all other treatments. Populations of S. Typhimurium LT2 (15-min drying) were reduced by 3.30 log CFU/mL with 70% ethanol, 5.38 log CFU/mL with SE1, and to non-detectable levels with 0.625% NaClO after 1 min, while S. Typhimurium ATCC 23564 (1-h drying) was non-detectable after 1 min by all treatments. Under soiled conditions, 10-min treatment with SE1 and 70% ethanol reduced both bacteria to non-detectable levels. Studies with concentrated switchgrass extractives combined with various other natural disinfectants or in hurdle approaches warrant further investigation.

Synergistic bactericidal effect of hot water with citric acid against Escherichia coli O157:H7 biofilm formed on stainless steel.

This study investigated the antimicrobial effect of hot water with citric acid against Escherichia coli O157:H7 biofilm on stainless steel (SS). Hot water (50, 60, or 70 °C) with 2% citric acid exhibited a synergistic bactericidal effect on the pathogen biofilm. It was revealed that hot water and citric acid combination induced sub-lethally injured cells. Additionally, mechanisms of the synergistic bactericidal effects of hot water with citric acid were identified through several approaches. In terms of biofilm matrix, hot water removes exopolysaccharides, a major component of extracellular polymeric substances (EPS), thereby increasing contact between surface cells and citric acid, resulting in a synergistic bactericidal effect. In terms of the cell itself, increased permeability of citric acid through cell membranes destructed by hot water promotes the inactivation of superoxide dismutase (SOD) in E. coli O157:H7, which induce synergistic generation of reactive oxygen species (ROS) which promote inactivation of cell by activating lipid peroxidation, resulting in destruction of the cell membrane. Therefore, it is interpreted that when hot water with citric acid is applied to E. coli O157:H7 biofilm, synergy effects on the biofilm matrix and cell itself have a complex interaction with each other, thus causing a dramatic synergistic bactericidal effect.

Assessment of microbial risk during Australian industrial practices for Escherichia coli O157:H7 in fresh cut-cos lettuce: A stochastic quantitative approach.

Escherichia coli O157:H7 risk associated with the consumption of fresh cut-cos lettuce during Australian industrial practices was assessed. A probabilistic risk assessment model was developed and implemented in the @Risk software by using the Monte Carlo simulation technique with 1,000,000 iterations. Australian preharvest practices yielded predicted annual mean E. coli O157:H7 levels from 0.2 to -3.4 log CFU/g and prevalence values ranged from 2 to 6.4%. While exclusion of solar radiation from the baseline model yielded a significant increase in concentration of E. coli O157:H7 (-5.2 -log fold), drip irrigation usage, exclusion of manure amended soil and rainfall reduced E. coli O157:H7 levels by 7.4, 6.5, and 4.3-log fold, respectively. The microbial quality of irrigation water and irrigation type both had a significant effect on E. coli O157:H7 concentrations at harvest (p < 0.05). The probability of illness due to consumption of E. coli O157:H7 contaminated fresh cut-cos lettuce when water washing interventions were introduced into the processing module, was reduced by 1.4-2.7-log fold (p < 0.05). This study provides a robust basis for assessment of risk associated with E. coli O157:H7 contamination on fresh cut-cos lettuce for industrial practices and will assist the leafy green industry and food safety authorities in Australia to identify potential risk management strategies.