Inactivation of Escherichia coli O157:H7 in dry-cured ham by high-pressure treatments combined with biopreservatives

The effect of high-pressure (HP) treatments combined with biopreservatives such as a commercial nisin or pediocin on the survival of Escherichia coli O157:H7 and the physicochemical, rheological and color characteristics of sliced dry-cured ham stored under mild temperature abuse conditions (8 °C) was investigated. Immediately after treatments, a synergistic antimicrobial effect was registered when 400 MPa and 500 MPa for 10 min combined with nisin were applied. After 60 d, this synergistic effect was only maintained with the combined treatment of 500 MPa and nisin. Counts of E. coli in dry-cured ham were not affected by either nisin or pediocin applied individually, whereas counts in pressurized samples were 3 log units lower than in non-treated dry-cured ham after 60 d of refrigerated storage. Changes in textural parameters caused by pressurization and biopreservatives were minor. Lightness (L*) values were slightly affected, with lower values in samples pressurized at 500 MPa in combination with bacteriocins and a trend to decrease during storage. Redness (a*) and yellowness (b*) were less modified by HP and biopreservatives and tended to diminish during refrigeration. The results obtained in the present work demonstrate the effectiveness of pressurization of sliced dry-cured ham at 500 MPa for 10 min combined with nisin on the inactivation of E. coli O157:H7.     

The antimicrobial effects and synergistic antibacterial mechanism of the combination of ε-Polylysine and nisin against Bacillus subtilis

The aim of the present study was to evaluate the combined effects of ε-Polylysine (ε-PL) and nisin and investigate the synergistic action of these compounds against Bacillus subtilis (B. subtilis).The combination of ε-PL and nisin showed synergistic anti-microbial activity against Escherichia coli (E. coli), B. subtilis and Staphylococcus aureus (S. aureus). SEM and TEM microscopy revealed that combined treatment with ε-PL and nisin synergistically damaged the morphology of tested bacterial cells. Propidium iodide (PI) infiltration experiments indicated that combined treatment with ε-PL and nisin synergistically enhanced the permeability of the bacterial cell membrane, likely reflecting the inhibition of both Na⁺K⁺- and Ca⁺⁺ Mg⁺⁺-ATPase activities through these compounds. The fluorescence spectrum showed an interaction between ε-PL and DNA, but not between nisin and DNA. The mode of ε-PL in binding with DNA was similar to that of ethidium bromide (EB). These results indicated that the uptake of ε-PL into cells was promoted through nisin, and subsequently, ε-PL interacted with the intracellular DNA achieving a synergistic effect.     

Inactivation of Listeria innocua in brined white cheese by a combination of nisin and heat

The objective of this study was to investigate the effect of nisin alone and in combination with heat (63 °C/5 min) on the inactivation of Listeria innocua in white cheese. Nisin was added at different concentrations (500, 1000, and 1500 IU ml⁻¹) to pasteurized milk before curd formation. The curd was soaked for 24 h in 10% solution of brine containing ca 10⁶ CFU ml⁻¹ of a cocktail mixture of three strains of L. innocua. Part of the nisin treated samples were heat treated at 63 °C/5 min. Total mesophilic count (TMC), L. innocua survivors and changes in the pH of white cheese were monitored each 2 d for a period of 12 d of storage at 4 or 10 °C. Nisin at 500 IU ml⁻¹ did not diminish TMC in white cheese compared to the control. The combination of heat and nisin (1000 or 1500 IU ml⁻¹) exhibited a bacteriostatic effect on TMC throughout the storage period at 4 or 10 °C. Nisin at 500 IU ml⁻¹ had a marginal inhibitory activity against L. innocua. However, nisin at 1000 and 1500 IU ml⁻¹l resulted in a more than 2 log₁₀ reduction in L. innocua count and the effect was more prominent at 10 °C. In comparison, the combination of nisin (1000 or1500 IU ml⁻¹) and heat treatment exhibited a synergistic inhibitory activity against L. innocua, where a complete elimination of the organism was accrued after 6 and 8 d of storage at 10 and 4 °C. Therefore, nisin and heat combination could be used as a prudent hurdle to preclude the growth of Listeria in white cheese, especially under the condition of abused refrigeration conditions.     

Evaluation of the factors affecting the activity of sakacin C2 against E. coli in milk

Sakacin C2 is a novel, broad spectrum bacteriocin secreted by Lactobacillus sake C2 isolated from Chinese traditional fermented cabbage. Effects of milk fat, emulsifiers, preservatives and homogenization on the activity of sakacin C2 against Escherichia coli ATCC 25922 in milk were evaluated by determined the changes of viable cell counts of E. coli ATCC 25922 after inoculation (10⁵ CFU/ml) then storage at 4 °C. Milk fat in pasteurized and homogenized milk products (low fat milk and whole milk) decreased the activity of sakacin C2 against E. coli ATCC 25922. Addition of 5 μl/ml of Tween 80 decreased the effect of sakacin C2 against E. coli ATCC 25922, but 5 mg/ml of lecithin increased the effect against E. coli ATCC 25922 in pasteurized and homogenized whole milk. Nisin and ε-Polylysine significantly increased the effect of sakacin C2 against E. coli ATCC 25922 in pasteurized and homogenized whole milk. Homogenization interfered with the activity of sakacin C2 against E. coli ATCC 25922 in pasteurized skim milk and whole milk. This study might lay the groundwork for the application of sakacin C2 as bio-preservative in milk and dairy products.     

Antibacterial activity of plant essential oils and extracts: The role of thyme essential oil,nisin, and their combination to control Listeria monocytogenes inoculated in minced fish meat

Food poisoning caused by Listeria monocytogenes leads to a 30% rate of mortality among patients. The antibacterial activity of cinnamon, thyme, and rosemary essential oils (EOs) and shallot and turmeric extracts was tested against L. monocytogenes using agar well and disc diffusion techniques. Results showed that thyme EO had the highest antimicrobial activity, followed by cinnamon and rosemary EOs, respectively. The antilisterial activity of thyme EO at 0.4%, 0.8%, and 1.2% levels, nisin at 500 or 1000 IU/g level, and their combination against L. monocytogenes was examined in minced fish samples. The antilisterial properties of nisin were also investigated in cooked minced fish treatments. Nisin at 500 or 1000 IU/g in the minced fish meat demonstrated bacteriostatic activity against L. monocytogenes. The use of thyme EO at 0.8% and 1.2% reduced the L. monocytogenes viable count below 2 log cfu/g after 6 days. Furthermore, simultaneous use of thyme EO at 0.8% and 1.2%, and nisin at 500 or 1000 IU/g level, reduced the L. monocytogenes viable count below 2 log cfu/g after the second day of storage. The antilisterial activity of nisin in the cooked minced fish samples was slightly stronger than that of the raw group.     

Inactivation of Escherichia coli O157:H7 by high pressure carbon dioxide combined with nisin in physiological saline,phosphate-buffered saline and carrot juice

The individual and combined effects of high pressure carbon dioxide (HPCD) and nisin (200 IU/mL) on the inactivation of Escherichia coli O157:H7 suspended in physiological saline (PS, pH 5.60), phosphate-buffered saline (PBS, pH 5.60 or 7.00) or carrot juice (pH 6.80) were evaluated. The pressure in this study was 5 and 8 MPa, the temperature was 25 °C–45 °C, and the treatment time was 5–65 min. Inactivation of cells in PS (pH 5.60) by HPCD followed first order kinetics, the k (the inactivation rates) increased while the D (decimal reduction time) decreased in the presence of nisin, however, the acid solution dissolving nisin rather than nisin itself played a prominent role in this combination effect with HPCD in PS buffer. The inactivation kinetics of cells in PBS (pH 5.60 or 7.00) and carrot juice (pH 6.80) by HPCD followed slow-to-fast two-stage kinetics and was fitted by the modified Gompertz equation. The M (the time at which the absolute death rate is maximum) significantly decreased in the presence of nisin. HPCD enhanced the sensitization of E. coli to nisin and the time for the complete inactivation was shortened by 2.5–5 min in PBS buffer and carrot juice by combination of HPCD and nisin (HPCD + nisin) than by HPCD alone. Regression coefficients (R²) and mean square error (MSE) were used to evaluate the model performance, indicating that the models could provide a good fitting to the experimental data.     

Combined effect of reuterin and lactic acid bacteria bacteriocins on the inactivation of food-borne pathogens in milk

Antimicrobial activity of reuterin in combination with different bacteriocins from lactic acid bacteria against food-borne pathogens in milk was investigated. A strong synergistic effect of reuterin in combination with nisin, lacticin 481 or enterocin AS-48 on Listeria monocytogenes was observed. Only nisin increased the antimicrobial activity of reuterin against Staphylococcus aureus. Bactericidal activity of reuterin towards Escherichia coli O157:H7, Salmonella enterica, Yersinia enterocolitica, Aeromonas hydrophila and Campylobacter jejuni was not enhanced significantly by the addition of any of the bacteriocins investigated. The synergism of reuterin and nisin against L. monocytogenes and S. aureus was also found at refrigeration temperatures, where the pathogens were completely inactivated. Refrigerated milk treated with both natural antimicrobials would mean a feasible system to control pathogenic contaminants.     

Combined effect of natural antimicrobials and thermal treatments on Alicyclobacillus acidoterrestris spores

Commercial fruit juices are currently pasteurized at temperatures between 80 °C and 100 °C and Alicyclobacillus acidoterrestris spores can survive these thermal treatments. A. acidoterrestris produces several chemical off-odour compounds which cause the fruit juice spoilage. The purpose of this research was to evaluate the effect of the combination of a thermal treatment with natural antimicrobials (nisin, citral and limonene) on the survival and recovery of A. acidoterrestris spores. Heat resistance determinations at 95 °C in pH 3.5 McIlvaine buffer with or without the addition of these antimicrobials (alone or combined) in the heating or recovery media were performed. None of the antimicrobials reduced the heat resistance of A. acidoterrestris spores when added to the heating medium. However, nisin (1.5 mg L⁻¹) alone was able to reduce the viable counts of this microorganism by more than two log cycles when added to the plating medium, even when no thermal treatment was previously applied. The different combinations of nisin with citral on the recovery medium, showed a strong synergistic effect on A. acidoterrestris. Limonene showed no inhibitory effect in either media (heating and recovery). The addition of natural antimicrobials such as nisin or citral, which would not affect the flavor or taste of citrus juices, alone or even combined, can help to the control of A. acidoterrestris.     

Combined effects of heat,acetic acid,and salt for inactivating Escherichia coli O157:H7 in laboratory media

Heat, acid, and salt have been commonly used to ensure the microbial safety of foods and are often used in combinations in many food products. When combined, they can produce different results, such as additive, synergistic, and antagonistic effects. However, there has been little investigation into the effect of these combination treatments. Therefore, in this study, the effect of combined treatment of heat, acid, and salt was investigated in laboratory media. All possible paired combinations among three factors, heat (55 °C), acid (0.25% acetic acid, [v/v]) and salt (3%, [w/v]) were tested and compared with individual treatments for killing E. coli O157:H7 in laboratory media. When salt was combined with heat, there was no significant difference in reduction of E. coli O157:H7 (additive effect). However, when acid was combined with heat, there was a higher reduction of E. coli O157:H7 (synergistic effect). When salt was combined with acid treatment, salt gave protection against acid treatment (antagonistic effect), thus, there was lower reduction of E. coli O157:H7 in the combined treatment than in the single acid treatment. Depending on the combination of preserving factors, results were different.     

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.     

Characterization and evaluation of lactic acid bacteria isolated from goat milk

The aim of this study was to characterise and select Lactic Acid Bacteria (LAB) from goat milk with potential probiotic use and to evaluate the safety of these cultures in artisanal cheeses. The isolates of LAB were subjected to simulation of tolerance to the gastrointestinal tract, haemolytic test, antimicrobial susceptibility, antibacterial activity, EPS production, gas production, evaluation of proteolytic activity, diacetyl production and tolerance to NaCl. The genus and species of the selected LAB isolates were confirmed using molecular identification. Three goat cheeses (1, 2 and control) were manufactured to evaluate the inhibitory action of LAB against Escherichia coli. Subsequently, all cheese samples underwent bacterial enumeration and physical–chemical analyses. Statistical analysis was performed. UNIVASF CAP 14 and 20 were differentiated by survival up to pH 2 and pancreatin, resistance to NaCl and antibacterial activity against Klebsiella pneumoniae. UNIVASF CAP 4 and 29 were characterised by resistance to intestinal juice and antibacterial activity against Salmonella Typhi and Listeria monocytogenes. UNIVASF CAP 27, 38, 43 and 139 exhibited diacetyl production, antibacterial activity against Bacillus cereus, Staphylococcus aureus and Enterococcus faecalis. UNIVASF CAP 35 and 138 were characterised by proteolytic activity, EPS production, antibacterial activity to E. coli and Shigella flexneri. A cocktail of these 10 isolates with potential probiotic properties were inoculated in artisanal goat cheese and improved microbiological safety of product against E. coli.     

Effects of blackberry juice on growth inhibition of foodborne pathogens and growth promotion of Lactobacillus

Berries such as blueberry, blackberry and raspberry possess several biological activities including antimicrobial and nutritional effects. In this study, the antimicrobial activities of blackberry (Rubus fruticosus) juice against foodborne pathogens including Listeria monocytogenes, Salmonella Typhimurium and Escherichia coli O157:H7 were investigated. Inhibition of growth of these foodborne pathogens was measured in broth (Luria–Bertani broth for E. coli O157:H7 and S. Typhimurium, and brain heart infusion broth for L. monocytogenes), skim milk and whole milk supplemented with 10% blackberry juice at different time points (0, 24, 48 and 72 h). The effects of blackberry juice on the growth of Lactobacillus casei, Lactobacillus plantarum and Lactobacillus rhamnosus were also investigated in Man–Rogosa–Sharpe (MRS) broth and skim and whole milk supplemented with blackberry juice. The growth of L. monocytogenes, S. Typhimurium and E. coli O157:H7 were significantly inhibited by blackberry juice by 1–3 logs in both milk and broth. We also observed that the growths of Lactobacillus strains were significantly stimulated (1–4 logs CFU/mL) by blackberry juice in both milk and MRS broth. These data clearly demonstrate that diluted blackberry juice can be used as a preservative in food processing and a preventive in foodborne infections as a natural antimicrobial.     

Antimicrobial activity of ascorbic acid alone or in combination with lactic acid on Escherichia coli O157:H7 in laboratory medium and carrot juice

The antimicrobial effects of ascorbic acid alone and in combination with lactic acid, against Escherichia coli O157:H7 in Brain Heart Infusion (BHI) broth and in carrot juice as a food model was investigated. In control samples, E. coli O157:H7 continued to growth from 3.98 ± 0.2 log CFU and reached to 8.88 ± 0.1 log₁₀ CFU after 8 h at 37° C; however, bacterial population was undetectable level in BHI in the presence of 0.4% ascorbic acid and 0.2% lactic acid. In carrot juice, E. coli O157:H7 continue to grow from initial population count of 4.41 ± 0.9 log₁₀ CFU to 8.75 ± 0.07 log₁₀ CFU at 37 °C for 24 h. Similarly, the bacteria population was undetectable when 0.2 or 0.4% ascorbic acid and 0.2% lactic acid applied. Our findings suggest the application of ascorbic acid, in combination with lactic acid, may have potential as preservative to inhibit the growth of E. coli O157:H7 in food.     

Antibacterial activity of bovine milk lactoferrin on the emerging foodborne pathogen Cronobacter sakazakii: Effect of media and heat treatment

Cronobacter sakazakii is a pathogen transmitted by food, with high osmotic resistance and tolerance to desiccation, which affects mainly to newborns, infants and immunocompromised adults. C. sakazakii infection in infants has been associated with consumption of powdered milk. The purpose of this study was to evaluate the antibacterial activity of native and iron-saturated bovine lactoferrin (bLF) (from 0.5 to 5 mg/ml) on non-desiccated and desiccated C. sakazakii (10⁴ CFU/ml) in different media (phosphate buffer, bovine skim milk and whey). In general, native bLF was the only effective form that inhibited growth of C. sakazakii in all media, its activity increasing with concentration and time of incubation. These results suggest that the antibacterial effect of bLF on C. sakazakii is mainly due to iron sequestration. However, iron-saturated bLF showed some effect by reducing the viability of C. sakazakii in whey. There has not been observed an increased sensitivity of desiccated bacteria to native bLF in phosphate buffer. However, although the antibacterial activity of native bLF against non-desiccated C. sakazakii was drastically reduced in milk or whey compared to phosphate buffer, there was a certain activity when it was assayed against desiccated cells in those media. The effect of some heat treatments on the antibacterial activity of native bLF was evaluated and only those of 72 °C for 15 s, 85 °C for 15 s, and 63 °C for 30 min maintained its whole activity.     

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.     

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.     

Radiation tolerance of Bacillus cereus pre-treated with carvacrol alone or in combination with nisin after exposure to single and multiple sub-lethal radiation treatment

Currently little information exists on the response of Bacillus cereus after repetitive exposure to combined mild treatments involving antimicrobials and gamma irradiation. Therefore, the aim of this present study was to evaluate the radiation stress on growth and physiology of B. cereus LSPQ 2872 vegetative cells at stationary phase, following exposure to single and repetitive sub-lethal γ-radiation treatment at 1 kGy simultaneously with carvacrol alone or combined with nisin at sub-inhibitory concentrations. Results indicated that the combination of carvacrol and nisin significantly enhance the radiation sensitivity of B. cereus since lower D₁₀ values were recorded after both single and repetitive irradiation treatments. Flow cytometric analysis of radiation-stressed B. cereus cells following repetitive treatment revealed the heterogenic behaviour of the bacterium leading to the induction of a radiation tolerance response. When compared to carvacrol alone, the combination of carvacrol and nisin developed also increased radioresistance if repetitively processed with γ-radiation at 1 kGy, since the decrease percentage of dead cells was accompanied by an increase in the number of injured cells. However, good agreement was not found between classical plate counting (log cfu reductions) and flow cytometry method. For both antimicrobials, the increase of radioresistance after repetitive mild treatment was not accompanied by augmentations of lag phase or growth rate. The structural changes of the outer membranes were assessed by TEM analysis and results revealed that radioresistance might be related to changes in the cell wall.     

Synergetic antibacterial efficacy of cold nitrogen plasma and clove oil against Escherichia coli O157:H7 biofilms on lettuce

This study aims to evaluate the antimicrobial effects of cold nitrogen plasma (CNP) and clove oil against Escherichia coli O157:H7 (E. coli O157:H7) biofilm on lettuce. Both clove oil (1 mg/mL, 2 mg/mL and 4 mg/mL) and CNP (400–600 W) displayed significant eradication effect on E. coli O157:H7 biofilms in vitro (p < 0.001). Subsequently, the antibiofilm effect of combined treatment was studied as well. Compared with the respective treatment, combined treatment exhibited remarkable synergistic effect on eradicating E. coli O157:H7 biofilms. The confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM) had also visually testified that the antibacterial effects of clove oil on E. coli O157:H7 biofilms (in vitro and on lettuce) were enhanced by CNP at 400 W for short treatment duration. The results of sensory evaluation indicated that combined treatment has mild negative effect on lettuce quality. Moreover, the synergetic antibacterial mechanism of clove oil and CNP against E. coli O157:H7 was concluded as that they could damage the bacterial cell wall and the outer membrane, leading to leakage of cellular components, such as nucleic acid and ATP.     

Effect of organic acids,hydrogen peroxide and mild heat on inactivation of Escherichia coli O157:H7 on baby spinach

Minimally processed baby spinach contaminated with Escherichia coli O157:H7 has been associated with multiple outbreaks of foodborne illnesses recently. Chlorinated water is widely used to wash vegetables commercially, but this washing procedure has limited efficacy and can lead to the formation of carcinogenic substances. This study was conducted to determine the effects of organic acids and hydrogen peroxide alone and in binary combinations with or without mild heat (40 and 50 °C) on the inactivation of Escherichia coli O157:H7 on baby spinach. Baby spinach leaves were dip-inoculated with E. coli O157:H7 to a level of 6 log CFU/g and stored at 4 °C for 24 h before treatment. Individual washing solutions (1% and 2% lactic acid [LA], citric acid [CA], malic acid [MA], tartaric acid [TA], acetic acid [AA], hydrogen peroxide [H₂O₂] as well as binary combinations of LA, CA, MA and H₂O₂ at final concentrations of 1% were used to decontaminate spinach leaves at 22, 40 or 50 °C for 2–5 min to test their efficacy in reducing E. coli O157:H7. Chlorinated water (200 ppm free chlorine) decreased the population of E. coli O157:H7 on baby spinach by only 1.2–1.6 log CFU/g, which was not significantly different from DI water washing. Washing with 1% LA at 40 °C for 5 min was the most effective treatment achieving a 2.7 log reduction of E. coli O157:H7 which is significantly higher than chlorine washing. Washing with LA + CA or LA + HP at 40 °C for 5 min was equally effective against E. coli O157:H7, resulting in a 2.7 log reduction of E. coli O157:H7. The application of mild heat significantly enhanced the efficacy of washing solutions on the inactivation of E. coli O157:H7. There was, however, no significant difference between treatments at 40 °C for 5 min and 50 °C for 2 min. The results suggested that the use of organic acids in combination with mild heat can be a potential intervention to control E. coli O157:H7 on spinach.     

Reuterin,lactoperoxidase, lactoferrin and high hydrostatic pressure on the inactivation of food-borne pathogens in cooked ham

The antimicrobial effect of high hydrostatic pressure (HHP) processing combined with reuterin, lactoperoxydase system (LPS) and lactoferrin (LF) on the survival of Listeria monocytogenes, Salmonella enterica subsp. enterica serovar Enteritidis and Escherichia coli O157:H7 in sliced cooked ham stored under strict refrigeration temperature (4 °C) and mild temperature abuse conditions (10 °C) was investigated. One day after treatment, L. monocytogenes counts in HHP at 450 MPa for 5 min were 0.8 log units lower, but a recovery was observed with counts not significantly different to those observed in control after 35 d. S. Enteritidis and E. coli O157:H7 levels were reduced around 5 log cfu/g by the pressure treatment (450 MPa/5 min) and the numbers of these pathogens did not increase significantly during the 35 d of storage at 4 °C. The individual application of reuterin and LPS influenced the survival of the three pathogens studied, extending the lag phase of L. monocytogenes and diminishing S. Enteritidis and E. coli levels throughout storage, whereas no effect was recorded when LF was added. When reuterin or LPS were applied in combination with HHP there was a synergistic antimicrobial effect against L. monocytogenes, avoiding at 4 °C the recovery observed with individual treatments. These combined treatments also kept the levels of S. Enteritidis and E. coli O157:H7 below the detection limit (<1 log unit) in cooked ham stored at 4 and 10 °C during 35 d. The results obtained in the present work suggest that HHP at 450 MPa for 5 min in combination with LPS or reuterin would be useful as a hurdle technology approach against L. monocytogenes, S. Enteritidis and E. coli O157:H7 in cooked ham.