Evaluation of a novel antimicrobial solution and its potential for control Escherichia coli O157:H7, non-O157:H7 shiga toxin-producing E. coli,Salmonella spp., and Listeria monocytogenes on beef

The goal of this study was to evaluate the efficacy of a novel antimicrobial solution made with chitosan, lauric arginate ester, and organic acids on Escherichia coli O157:H7, Salmonella spp., Listeria monocytogenes, and non-O157 shiga toxin-producing E. coli cocktails and to test its potential to be used as a marinade for raw beef. Fresh beef top round steaks were surface-inoculated with the pathogen cocktails at approximately 2.5 or 4.5 Log CFU/cm², marinated with the antimicrobial solution (AMS), and then stored at 4 °C for 6, 24, and 48 h. Three commercially available marinades were used for comparison. Results revealed that AMS had the most antimicrobial effect regardless of the type or inoculation level of pathogens (P < 0.05). After 6 h, the AMS marination reduced all pathogens to levels below the limit of detection (<1 Log CFU/cm²), resulting in a 3.5 Log CFU/cm² reduction. When AMS was diluted with autoclaved distilled water by 5 times (AMS 1:5) or 10 times (AMS 1:10), its antimicrobial efficacy was impacted by marination time, the inoculated pathogens, and the inoculation levels. This study demonstrates that the developed antimicrobial solution has a great potential to be used during marination by consumers to ensure better food safety.     

Simulation of decontamination and transmission of Escherichia coli O157:H7, Salmonella Enteritidis,and Listeria monocytogenes during handling of raw vegetables in domestic kitchens

Epidemiological data indicates that a large number of foodborne illnesses are attributed to cross-contamination during food preparation in the domestic kitchen. The objectives of this study were to evaluate the efficiency of household washing practices in removing Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella Enteritidis on artificially contaminated lettuce and to determine the transfer rate of these three foodborne pathogens from contaminated lettuce to wash water, tomato, cabbage, and cutting boards during washing and cutting processes. Washing under the running tap water with scrubbing for 60 s was the most effective method in reducing pathogen populations by 1.86–2.60 log₁₀ CFU/g. Also, final rinsing and scrubbing practices were found to enhance the efficiency of washing treatment. In this study, the transfer rates of S. Enteritidis, E. coli O157:H7, and L. monocytogenes from cutting board to cabbage and tomato via cutting process (17.5–31.7%) were higher (P < 0.05) than from wash water to cabbage and tomato (0.8–23.0%) during washing treatment. Overall, our findings suggest that wash water and cutting board can be potential vehicles in the dissemination of foodborne pathogens. Therefore, there is a need to promote consumer awareness for proper handling practices in the kitchen to minimise the risk of foodborne infection.     

Ralstonia insidiosa serves as bridges in biofilm formation by foodborne pathogens Listeria monocytogenes,Salmonella enterica,and Enterohemorrhagic Escherichia coli

Biofilm formation on abiotic surfaces in fresh produce processing facilities may play a role in foodborne outbreaks by providing protective microniches for pathogenic bacteria. Our previous study showed that a strain of Ralstonia insidiosa isolated from a fresh produce processing plant could enhance the incorporation of Escherichia coli O15:H7 in biofilms under various environmental conditions. These results raised the concern that R. insidiosa might have the ability to incorporate other foodborne pathogens and promote their survival and growth in biofilms. To test this hypothesis, 6 strains of Shiga toxin producing E. coli, 2 strains of Salmonella, and 6 strains of Listeria monocytogenes were examined for dual-species biofilm formation with R. insidiosa. A significant increase in biomass formation was observed in 7 of the 14 R. insidiosa-pathogen combinations, while significantly enhanced incorporation of pathogenic cells into biofilms was seen in 12 of the 14 R. insidiosa-pathogen combinations. The synergistic interactions between R. insidiosa and the tested foodborne pathogens seemed dependent on intimate cellular contact between the two strains. Overall, this study showed that R. insidiosa could enhance the incorporation of biofilms of different types of foodborne pathogenic bacteria and should be considered a bridging bacterium for biofilm formation in various food processing environments.     

Antimicrobial activity of mustard essential oil against Escherichia coli O157:H7 and Salmonella typhi

The aim of this study was to investigate how mustard essential oil (EO) affected the cell membrane of Escherichia coli O157:H7 and Salmonella typhi. Intracellular pH and ATP concentration and the release of cell constituents were measured when mustard EO was in contact with E. coli and S. typhi at its minimal inhibitory concentration (MIC) and maximal tolerated concentration (MTC). The treatment with mustard EO affected the membrane integrity of bacteria and induced a decrease of the intracellular ATP concentration. Also, the extracellular ATP concentration increased and a reduction of the intracellular pH was observed in both bacteria. A significantly (P ? 0.05) higher release of cell constituent was observed when both bacteria cells were treated with mustard EO. Electronic microscopy observations showed that the cell membranes of both bacteria were apparently damaged by mustard EO. In conclusion, mustard EO affects the concentration of intracellular component, such as ATP in both bacteria and affects the pH suggesting that cytoplasmic membrane is involved in the antimicrobial action of mustard EO. Mustard EO can be used as an effective antimicrobial agent. We have demonstrated that mustard EO affected the cell membrane integrity, resulting in a loss of cell homeostasis.     

Detection of viable but non-culturable Escherichia coli O157:H7 from vegetable samples using quantitative PCR with propidium monoazide and immunological assays

Quantitative differentiation of the live fraction of pathogens in raw food samples is highly critical from a public health risk perspective, as many studies have shown that under stress conditions major foodborne pathogens enter a viable but non-culturable (VBNC) state in which bacteria can remain for long periods of time and maintain the potential for virulence. The objective of this study was to evaluate the applicability of propidium monoazide (PMA) quantitative PCR (qPCR) and immunological methods for detection of Escherichia coli O157:H7 VBNC populations induced by low temperature on the surface of lettuce and spinach plants. The primer/probe set selected influenced the qPCR signal in mixtures with a defined ratio of viable and non-viable cells. The PMA qPCR used in a background of added dead pathogens and epiphytic bacteria gave a detection limit of 10³ CFU/g leaf and a linear quantitative detection range of 5 log. During quantification of VBNC cells from lettuce and spinach samples there was a good correlation between the PMA qPCR results and viable counts detected by microscopy, showing that PMA qPCR gives an accurate indication of the VBNC population. However, the commercially available immunoassay methods used to detect Shiga-like toxin production and the O157 antibody in vegetable samples with no detectable culturable pathogen underestimated the number of samples contaminated with E. coli O157:H7 VBNC cells. Results indicate that PMA qPCR is a suitable technique for the detection and quantification of VBNC cells of foodborne pathogens in contaminated raw lettuce and spinach.     

Effect of hydrogen peroxide vapor treatment for inactivating Salmonella Typhimurium,Escherichia coli O157:H7 and Listeria monocytogenes on organic fresh lettuce

In this study, the efficacy of hydrogen peroxide vapor (HPV) for reducing Salmonella Typhimurium, Escherichia coli O157:H7 and Listeria monocytogenes on lettuce was investigated as well as its effect on lettuce quality. Lettuce was inoculated with a cocktail containing three strains of each pathogen then treated with vaporized hydrogen peroxide for 0, 2, 4, 6, 8 and 10 min. The concentrations of hydrogen peroxide used were 0, 1, 3, 5 and 10%. With increasing treatment time and hydrogen peroxide concentration, HPV treatment showed significant (P < 0.05) reduction compared to the control (0%, treated with vaporized distilled water). In particular, vaporized 10% hydrogen peroxide treatment for 10 min was the most effective combination for reducing the three pathogens on lettuce. The reduction levels of S. Typhimurium, E. coli O157:H7 and L. monocytogenes on lettuce were 3.12, 3.15 and 2.95 log₁₀ CFU/g, respectively. Furthermore, there were no significant (P > 0.05) quality changes (color and texture) of lettuce among all tested samples, and hydrogen peroxide residues were not detected after 36 h storage time in any of the treated samples. These results suggest that HPV treatment could be an alternative method for reducing S. Typhimurium, E. coli O157:H7 and L. monocytogenes on fresh produce.     

Addition of phytochemical-rich plant extracts mitigate the antimicrobial activity of essential oil/wine mixtures against Escherichia coli O157:H7 but not against Salmonella enterica

Marinades for preparing raw meats for cooking are frequently made of wine and herbs. We simulated several formulations of potential antimicrobial marinades with these components and other food compatible/food derived extracts. Red wine formulations containing essential oils from oregano or thyme, or their primary active components carvacrol and thymol, respectively, and a mixture of plant extract powders from phytochemical-rich apple skin, green tea, and olive, were evaluated for inhibitory activity against the foodborne pathogens Escherichia coli O157:H7 and Salmonella enterica. Red wine alone exhibited low activity, as did the plant extract suspended in the wine. Surprisingly, the high activity of oregano or thyme essential oils in red wine was reduced in E. coli, but not in Salmonella, by addition of the plant extract. This study shows that essential oils in red wine can be an effective antimicrobial in food, however the possibility exists that phytochemicals, added to the treatment solution or natively present in the food itself, could adversely impact the antimicrobial activity and should be addressed with future studies.     

Efficacy of the combined application of oregano and rosemary essential oils for the control of Escherichia coli,Listeria monocytogenes and Salmonella Enteritidis in leafy vegetables

This study assessed the effect of the combined application of essential oils (EOs) from Origanum vulgare L. – oregano (OVEO) and Rosmarinus officinalis L. – rosemary (ROEO), alone or in combination at subinhibitory concentrations, against three pathogenic bacteria that are associated with fresh leafy vegetables: Listeria monocytogenes (L. monocytogenes), Escherichia coli (E. coli) and Salmonella enterica Serovar Enteritidis (S. Enteritidis). The inhibitory effects were evaluated by determining the minimum inhibitory concentration (MIC) and the fractional inhibitory concentration index (FICI) and assessing the viable cell counts in vegetable broth and artificially infected vegetables over time. Still, the effects of the EOs on native spoilage native flora were assessed. The MIC of OVEO was 0.6 μL/mL against the test strains either in single and mixed inoculum. The MIC of ROEO was 5 μL/mL against L. monocytogenes and E. coli and 10 μL/mL against S. Enteritidis in single inocula, whereas it was 10 μL/mL against the mixed inoculum. The FICI of the combined EOs was 0.5 against the mixed bacterial inoculum, which suggested a synergic interaction. The incorporation of OVEO and ROEO alone (MIC) or combined at different subinhibitory concentrations in vegetable broth resulted in a decrease in the viable cell counts of all test strains over 24 h. Similarly, the EOs alone or in the tested combinations reduced the viable cell counts of all test strains in experimentally infected fresh vegetables, besides to decrease the counts of spoiling native flora (mesophilic bacteria, enterobacteria and fungi). These findings reinforce the rational for the use of OVEO and ROEO in combination at subinhibitory concentrations to guarantee the safety and extend the shelf life of fresh vegetables.     

Antimicrobial effects of modified chitosan based coating containing nanoemulsion of essential oils,modified atmosphere packaging and gamma irradiation against Escherichia coli O157:H7 and Salmonella Typhimurium on green beans

The antibacterial activity of modified chitosan-based coatings containing nanoemulsion of essential oils (EOs), gamma irradiation, modified atmosphere packaging (MAP), alone or in combinations, against Escherichia coli O157:H7 and Salmonella Typhimurium was evaluated on inoculated green bean samples. Firstly, four different nanoemulsions, made of carvacrol, mandarin, bergamot and lemon Eos, respectively, were compared in terms of minimum inhibitory concentration (MIC) against the two bacteria evaluated in vitro using the micro-broth dilution method. Carvacrol nanoemulsion resulted to be the most effective antibacterial agent and was therefore selected to be incorporated into modified chitosan (MC) to form a bioactive coating. Secondly, the radiosensitivity of E. coli and S. Typhimurium to gamma irradiation was evaluated on inoculated green beans after coating deposition and MAP. Results showed that, without MAP, MC-based coating containing carvacrol nanoemulsion significantly increased the radiosensitization of E. coli and S. Typhimurium by 1.32-fold and 1.30-fold, respectively. Remarkably, the use of bioactive coating under MAP caused a synergistic effect with an increase in radiosensitivity by 1.80-fold and 1.89-fold for E. coli and S. Typhimurium, respectively. Thirdly, the antibacterial effects of the antimicrobial coating, gamma irradiation, MAP alone and their combinations were evaluated against these two bacteria during a 13-days storage of green beans at 4 °C. Bioactive coating deposition or gamma irradiation treatment resulted effective in controlling the growth of the two bacteria during the entire shelf-life. Moreover, it was also found that the combined treatment of antimicrobial coating, gamma irradiation and MAP caused the reduction of microbial population to undetectable levels during the whole storage period for E. coli and from day 7 to the end of storage for S. Typhimurium. The obtained results can be interested to food companies aiming to ensure the food safety with a prolonged shelf life.     

Simultaneous detection of Escherichia coli O175:H7, Salmonella spp., and Listeria monocytogenes by multiplex PCR

The wide application of nucleic acid amplification techniques and the increasing industrial interest toward rapid methods has led to the development and application of PCR based methods for the detection of microbial pathogens in food. In the present paper we describe the development of a multiplex PCR method for simultaneous detection of Salmonella enterica serovar Typhimurium, Listeria monocytogenes and Escherichia coli O157:H7 in a complex food matrix (liquid whole egg).Four different DNA extraction procedures were evaluated for their application on food and, among these, Chelex resin combined with a DNA purification step were found to better perform on the food system considered.A multiplex PCR system was developed, based on the evaluation and combination of published primer sets, and applied to the simultaneous detection of the target pathogens plus an internal amplification control, both in culture media and in a model food system.The overall system proposed, based on an overnight enrichment step followed by DNA isolation and multiplex PCR, was satisfactorily tested for its specificity and sensitivity and allowed the detection of the presence of bacterial DNA and the identification of the target pathogens down to 10 cells/25 g liquid whole egg.     

Antimicrobial activity of malic acid against Listeria monocytogenes,Salmonella Enteritidis and Escherichia coli O157:H7 in apple,pear and melon juices

Minimal inhibitory (MIC) and minimal bactericidal (MBC) concentrations of malic acid against Listeria monocytogenes, Salmonella Enteritidis and Escherichia coli O157:H7 inoculated in apple, pear and melon juices stored at 5, 20 and 35 °C were evaluated. MICs and MBCs against L. monocytogenes, S. Enteritidis and E. coli O157:H7 were significantly affected by storage temperature, juice characteristics and type of microorganism. Malic acid was more effective at 35 and 20 °C than at 5 °C in all studied fruit juices. E. coli O157:H7 was more resistant to malic acid than S. Enteritidis and L. monocytogenes. Apple, pear and melon juices without malic acid were inhibitory to E. coli O157:H7, S. Enteritidis and L. monocytogenes at 5 °C, whereas, MBCs of 1.5% (v/v) of malic acid in apple and pear juices, and 2% (v/v) in melon juice at 5 °C were needed to reduce E. coli O157:H7, those concentrations being higher than those required to reduce S. Enteritidis and L. monocytogenes in those fruit juices. In addition, concentrations of 2%, 2.5% and 2.5% (v/v) of malic acid added to apple, pear and melon juices, respectively, were required to inactivate the three pathogens by more than 5 log cycles after 24 h of storage at 5 °C. Transmission electron microscopy showed that malic acid produced damage in the cell cytoplasm of pathogens without apparent changes in the cell membrane.     

Antimicrobial activity of essential oils against Escherichia coli O157:H7 in organic soil

Soil can be a significant source of preharvest contamination of produce by pathogens. Demand for natural pesticides such as essential oils for organic farming continues to increase. We examined the antimicrobial activity of several essential oils against Escherichia coli O157:H7 in soil. Two essential oils (cinnamaldehyde and eugenol), two bio-pesticides (Ecotrol and Sporan) containing essential oils, and an organic acid (acetic acid) at 0.5%, 1.0%, 1.5% and 2.0%, were mixed with organic sandy soil and inoculated with five different strains of E. coli O157:H7. Soils were incubated at room temperature (22 °C) and samples obtained at 1, 7 and 28 days were enumerated to determine survival. E. coli O157:H7 populations in soil were reduced by up to 5 log cfu/g after 24 h incubation at room temperature with 2% cinnamanaldehyde, Ecotrol, Sporan or vinegar. Reduction in E. coli O157:H7 by eugenol was not significantly different from control. Overall, E. coli O157:H7 strain 4406 was the most sensitive of all the five strains tested and cinnamaldehyde was superior to other treatments in reducing E. coli O157:H7 in soil. In general, increases in essential oil concentrations corresponded to reduced survival of E. coli O157:H7 with all oils used in this study. The results suggest that oils can reduce potential contamination of fresh organic produce inadvertently contaminated by soil.     

Antibacterial activity and mechanism of cinnamon essential oil against Escherichia coli and Staphylococcus aureus

Cinnamon essential oil (EO) exhibited effective antibacterial activity against foodborne spoilage and pathogenic bacteria in model systems using Escherichia coli and Staphylococcus. The minimum inhibition concentration (MIC) of cinnamon EO was similar for both bacteria (1.0 mg/ml) while the minimum bactericide concentration (MBC) were 4.0 mg/ml and 2.0 mg/ml for E. coli and Staphylococcus aureus. GC–MS analysis confirmed that cinnamaldehyde was the major constituent in cinnamon EO (92.40%). Much effort was focused on elucidating the mechanism of antibacterial action of cinnamon EO against E. coli and S. aureus by observing the changes of cell microstructure using scanning electron microscope, determination of cell permeability, membrane integrity and membrane potential. After adding cinnamon EO at MIC level, there were obvious changes in the morphology of bacteria cells indicating cell damage. When cinnamon EO were added at MBC levels, the cells were destroyed. Cinnamon EO led to leakage of small electrolytes, causing rapid increase in the electric conductivity of samples at the first few hours. The values for E. coli and S. aureus reached 60% and 79.4% respectively at 7 h. Moreover, the concentration of proteins and nucleic acids in cell suspension also rose with increased cinnamon EO. Bacterial metabolic activity was decreased 3–5 folds as reflected by the results of membrane potential. Overall, S. aureus was more susceptible to cinnamon EO than E. coli.     

A comparison of saturated steam and superheated steam for inactivation of Escherichia coli O157:H7, Salmonella Typhimurium,and Listeria monocytogenes biofilms on polyvinyl chloride and stainless steel

This study was performed to compare the effectiveness of saturated steam (SS) and superheated steam (SHS) in the inactivation of Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes biofilms on polyvinyl chloride (PVC) and stainless steel. Biofilms were formed on PVC and stainless steel coupons by using a mixture of three strains each of three foodborne pathogens at 25 °C. After biofilm development, PVC and stainless steel coupons were treated with SS at 100 °C and SHS at 125, 150, 175, and 200 °C for 5, 10, 20, and 30 s on both sides. The viable cell numbers of biofilms were significantly (p < 0.05) reduced as SHS temperature and exposure time increased. For all biofilm cells, SHS treatment resulted in an additional log reduction compared to SS treatments. After exposure to 200 °C steam for 30 s or 10 s on PVC or stainless steel, respectively, the numbers of biofilm cells were reduced to below the detection limit (1.48 log). This study demonstrated that SHS treatment effectively reduced populations of biofilm cells and reduced disinfection time compared to SS treatments and further evaluated its potential as an excellent intervention for controlling microbial biofilms and enhancing safety in the food processing industry.     

Quantitative detection of viable foodborne E. coli O157:H7, Listeria monocytogenes and Salmonella in fresh-cut vegetables combining propidium monoazide and real-time PCR

The increase of foodborne outbreaks associated with fresh vegetables has highlighted the importance of developing rapid and specific methods for the detection and quantification of foodborne pathogens. In this sense, real-time PCR (qPCR) fulfills these requirements although it may detect dead cells. Recently, a potential strategy to specifically detect viable cells has been proposed relying on the use of DNA binding molecules as sample pretreatment previous to the qPCR. In this study propidium monoazide (PMA) and reagent D, combined with qPCR, were evaluated for the detection and quantification of viable Escherichia coli O157:H7, Salmonella and Listeria monocytogenes. Initially, the optimal concentration of both reagents was determined for discrimination between viable and dead bacteria in cell suspensions. Although both reagents showed similar reductions for the three pathogens, reagent D was toxic to L. monocytogenes and Salmonella and therefore only PMA was used to evaluate the applicability of this technique on food samples. A final concentration of 50 μM PMA was assayed in artificially inoculated spinach and mixed salad. PMA-qPCR signal was negative for all dead cell concentrations tested except for mixed salad inoculated with L. monocytogenes at the highest concentration. These results demonstrate that PMA-qPCR is a suitable technique for the detection and quantification of viable pathogens in fresh-cut vegetables at the levels normally found in vegetable samples.     

Efficacy of sanitizers in reducing Escherichia coli O157:H7, Salmonella spp. and Listeria monocytogenes populations on fresh-cut carrots

Shredded carrots were inoculated with Escherichia coli O157:H7, Salmonella or Listeria monocytogenes and washed for 1 or 2 min with chlorine (Cl; 200 ppm), peroxyacetic acid (PA; 40 ppm) or acidified sodium chlorite (ASC; 100, 200, 500 ppm) under simulated commercial processing conditions. After washed, the carrots were spin dried, packaged and stored at 5 °C for up to 10 days. Bacterial enumeration was significantly (P ⩽ 0.05) reduced by 1, 1.5 and 2.5 log CFU/g after washing with ASC 100, 250 and 500 ppm, respectively. All sanitizers reduced pathogen load below that of tap water wash and unwashed controls. During storage at 5 °C the bacterial load of all treatments increased gradually, but to different extent in different treatments. ASC inhibited bacterial growth more effectively than the other sanitizers and also maintained the lowest pathogen counts (<1 log CFU/g) during storage. Organic matter in the process water significantly (P ⩽ 0.05) reduced the antibacterial efficacy of Cl, but not that of PA or ASC. Therefore, ASC shows the potential to be used as a commercial sanitizer for washing shredded carrots.     

Antimicrobial activities of red wine-based formulations containing plant extracts against Escherichia coli O157:H7 and Salmonella enterica serovar Hadar

We evaluated the antimicrobial activities of three red wine based plant extract/plant essential oil (EO) formulations: olive extract powder/oregano EO, apple skin extract powder/lemongrass EO, and green tea extract powder/bitter almond EO, and their formula permutations against the foodborne pathogens Escherichia coli O157:H7 and Salmonella enterica serovar Hadar. The following parameters that were expected to affect antimicrobial activity were evaluated: pH (3.6 or 7.0), temperature (4, 22, and 37 °C), incubation time (0, 5, 30, and 60 min) and bacterial load (∼10⁴ or ∼10⁹ bacteria/ml). The formulations with oregano, lemongrass, or bitter almond EO inhibited the growth of both pathogens at acidic pH with bacterial loads of ∼10⁴ or ∼10⁹ bacteria/ml. At neutral pH, the formulations were less effective. Although all the 3-component formulations were fast acting; showing significant activity in less than 5 min, only the activity of the lemongrass-containing formulation markedly increased with longer incubation times from 0 to 60 min. Activity also increased for all 3-component formulations with increasing temperature from 4 °C to 37 °C, although the activity of the lemongrass formulation appeared to level off at 22 °C. Of the tested formulations, the lemongrass EO formulations, at pH 3.6, appeared to be the most effective against the tested pathogens, especially against Salmonella. The most active formulations merit evaluation for antimicrobial efficacy in liquid and solid food.     

Reduction of Escherichia coli O157, Listeria monocytogenes and Salmonella enterica sv. Typhimurium populations on fresh-cut bell pepper using gaseous ozone

Bacterial contamination is the main cause of food poisoning which can lead to diarrhoea, abdominal cramp, vomiting as well as death. Bacterial contamination can potentially be controlled by application of gaseous ozone due to its antibacterial activity. Therefore, the objective of this study was to investigate the efficacy of gaseous ozone for reducing food-borne pathogens such as Escherichia coli O157, Listeria monocytogenes and Salmonella enterica sv. Typhimurium on fresh-cut bell pepper. Efficacy of gaseous ozone to reduce bacterial populations was investigated in vitro and in vivo. Results showed that optimum effect of ozone on reducing bacterial populations was achieved with short term exposures of less than 6 h. Ozone reduced the bacterial population by disrupting bacterial cell structure, which lead to cellular death. Results also showed that bacterial cells have different resistance to ozone where ozone was more effective against L. monocytogenes, followed by E. coli O157 and Salmonella Typhimurium. Optimal reduction of the bacterial population on fresh-cut bell pepper was achieved with exposure to 9 ppm ozone for 6 h. This treatment reduced 2.89, 2.56 and 3.06 log of E. coli O157, Salmonella Typhimurium and L. monocytogenes populations, respectively. In conclusion, exposure to 9 ppm ozone for 6 h helps in reducing food-borne pathogen on fresh-cut bell pepper and has high potential to be an alternative sanitization treatment to reduce pathogen population on fruit.     

The bactericidal activity of acidic electrolyzed oxidizing water against Escherichia coli O157:H7, Salmonella Typhimurium,and Listeria monocytogenes on raw fish,chicken and beef surfaces

The bactericidal efficacy of acidic electrolyzed oxidizing water (AC-EW) (pH = 2.30, free chlorine = 38 ppm) and sterile distilled water (DW) on three pathogens (Escherichia coli O157:H7 Salmonella Typhimurium, and Listeria monocytogenes) inoculated on raw trout skin, chicken legs and beef meat surfaces was evaluated. The decontaminating effect of AC-EW and DW was tested for 0 (control), 1, 3, 5 and 10 min at 22 °C. AC-EW significantly (P < 0.05) reduced the three pathogens in the inoculated samples compared to the control and DW. The level of reduction ranged between ca.1.5–1.6 logs for E. coli O157:H7 and S. Typhimurium in the inoculated foods. However, AC-EW exhibited less bactericidal effect against L. monocytogenes (1.1–1.3 logs reduction). AC-EW elicited about 1.6–2.0 log reduction in the total mesophilic count. Similar treatment with DW reduced pathogens load by ca. 0.2–1.0 log reduction and total mesophiles by ca. 0.5–0.7 logs. No complete elimination of the three pathogens was obtained using AC-EW possibly because of the level of organic matter and blood moving from food samples to the AC-EW solution. This study demonstrates that AC-EW could considerably reduce common foodborne pathogens in fish, chicken and beef products.     

Quantification of Salmonella spp., Listeria monocytogenes and Escherichia coli O157:H7 in non-spiked food products and evaluation of real-time PCR as a diagnostic tool in routine food analysis

Escherichia coli O157:H7, Listeria monocytogenes and Salmonella spp. are foodborne pathogens frequently associated with foods such as poultry, ready-to-eat products, fruits and vegetables. PCR-based procedures are rapid, sensitive and accurate; in particular, real-time PCR (qPCR), which besides being an automated high-throughput technique, allows quantification of foodborne pathogens. In the present work, qPCR-based methods were applied for the quantitative detection of E. coli O157:H7, Salmonella spp. and L. monocytogenes in a total of 306 non-spiked food samples in a study carried out in two laboratories simultaneously. qPCR allowed the detection of the three pathogens in around 20% of the analyzed samples for each pathogen. Quantification results revealed the presence of the three pathogens mostly at levels between 10² and 10⁴ cells/g. Besides quantification, the qPCR results (presence/absence) were compared with those of the standard mini-VIDAS system. In order to determine which were the “true” positive samples, conventional PCR was carried out after the corresponding enrichment for each pathogen. These results were considered as the gold standard for further analysis. The statistical analysis of global data recording the presence of E. coli O157:H7 and L. monocytogenes, together with data previously obtained for Salmonella spp., revealed that qPCR outperformed the mini-VIDAS procedures, in terms of both time and accuracy. Thus, these results proved qPCR to be useful as a rapid diagnostic test for the direct detection of pathogens in food, without the need for enrichment steps.