Virucidal effect of high power ultrasound combined with a chemical sanitizer containing peroxyacetic acid for water reconditioning in the fresh-cut industry

The use of chlorine-based sanitizers is widespread throughout the fresh produce industry in the process water to maintain microbial safety of produce, avoid cross-contamination and recycle water. In this study, alternative disinfection technologies were investigated due to the negative reports showing chlorine instability in the presence of organic matter and the undesirable by-product residues. Tsunami^® (15.2% peroxyacetic acid and 11.2% hydrogen peroxide) and high-power ultrasound (HPU, 0.56 kW/l, 20 kHz), alone or in combination, were evaluated in lettuce process water analyzing their ability to reduce MNV during either produce washing or water recycling. PAA concentrations of 6.4, 12.8 and 16 mg/l as well as HPU were insufficient for application in the produce washing tank where a rapid MNV inactivation is required. In contrast, a complete rapid inactivation of MNV (7.4 log TCID₅₀) was recorded when 80 mg/l of PAA was used. For recycled water reconditioning, HPU was applied and no virus reduction was observed. Two PAA concentrations (6.4 and 12.8 mg/l) that can be found in the recycled water were also tested. The MNV inactivation was faster at 12.8 mg/l than at 6.4 mg/l of PAA with 2.6 and 2 log reductions after 20 min contact. One additional log reduction was achieved 20 and 40 min later, respectively. MNV inactivation fitted the Bipashic model well. A contact time of 95 and 115 min would be required to obtain the complete elimination of MNV (4.4 log TCID₅₀) by 6.4 and 12.8 mg/l of PAA. PAA efficacy was not enhanced by the combination with HPU in contrast to the inactivation of pathogenic bacteria tested in other studies. This indicates that viruses are more resistant to this hurdle technology than pathogenic bacteria such as Escherichia coli O157:H7, Salmonella and other HPU conditions such as higher frequencies and combination of HPU with other physical treatments need to be explored.     

Inactivation of Escherichia coli O157:H7 in vitro and on the surface of spinach leaves by biobased antimicrobial surfactants

This study was conducted to evaluate the effect of biosurfactants on the populations of Escherichia coli O157:H7 in suspension and on spinach leaves. Eight surfactants including four soybean oil-based biosurfactants, sodium dodecyl sulfate (SDS), polyoxyethylene sorbitan monooleate (Tween 80), sophorolipid (SO) and thiamine dilauryl sulfate (TDS) at concentrations of 0.1%, 0.5% and 1.0% were tested in bacterial suspension, and the most effective biosurfactants were applied on spinach leaves. Results showed that the soybean oil-based biosurfactants, SDS or Tween 80 did not significantly affect E. coli O157:H7 populations. SO and TDS at concentrations of 1.0% were effective in reducing E. coli O157:H7 populations in bacterial suspension. E. coli O157:H7 with an initial population of 7.1 log CFU/mL was not detectable (detection limit: 1 log CFU/mL) after 1 min in 1.0% TDS or after 2 h in 1.0% SO. On spinach leaves, SO at 1% did not significantly affect E. coli when compared to a water wash during 7 days post-treatment storage at 4 °C. However, TDS (1.0%) wash was as effective as 200 ppm chlorine in reducing population of spot inoculated E. coli O157:H7, achieving 3.1 and 2.7 log CFU/per leaf at day 0, and 1.4 and 1.9 log CFU/leaf at day 7 when compared with a water wash. No apparent change in spinach visual quality was observed. None of treatments caused changes in visual quality of spinach. Electron micrographs suggested ultrastructural damage of bacterial cells such as separation of the outer membrane from the cytoplasmic membrane. Overall, our results showed that SO and TDS may be potential sanitizers in inactivating human pathogens such as E. coli O157:H7 in wash water and on fresh produce.     

Characteristics of coliphage ECP4 and potential use as a sanitizing agent for biocontrol of Escherichia coli O157:H7

Escherichia coli O157:H7 is an important pathogenic bacterium to humans because it produces various toxins, such as shiga-toxin. Coliphage ECP4, which belongs to the Siphoviridae family, was isolated from bovine feces to test its utility as a potential agent for the biocontrol of E. coli O157:H7. The burst size of coliphage ECP4 was about 80 PFU/cell, after a latent period of 30–35 min. Coliphage ECP4 was susceptible to temperatures above 70 °C; however, its stability was slightly reduced to 1–2 log PFU/ml after 30 min in 70% ethanol. In addition, the shiga toxin gene was not detected on coliphage ECP4. Coliphage ECP4 inhibited the growth of E. coli O157:H7 in vegetable juice, and was not detected in cabbage after 5 h. When coliphage ECP4 was applied to biofilm-formed E. coli O157:H7, E. coli O157:H7 was efficiently reduced. The newly identified coliphage ECP4 might effectively reduce E. coli O157:H7 or its biofilmed-form. Therefore, the coliphage ECP4 might be an efficient sanitizer for fresh produce contaminated with E. coli O157:H7 in the biofilm environment.     

Reduction of Escherichia coli O157:H7 and Salmonella Typhimurium DT 104 on fresh produce using an automated washer with near neutral electrolyzed (NEO) water and ultrasound

Automated produce washers can be a useful processing aid when treating fresh produce contaminated with pathogens. The use of near neutral pH electrolyzed (NEO) water as a wash or sanitizing solution has been shown to lead to significant reductions of Escherichia coli O157:H7 and Salmonella on fresh produce. To further enhance reported pathogen reductions, the effects of a combined NEO water (155 mg/L free chlorine, pH 6.5) and ultrasound wash protocol on lettuce and tomatoes inoculated with E. coli O157:H7 and S. Typhimurium DT 104 were studied. The effects of the pH of NEO water and washer agitation on pathogen reductions were also assessed. Inoculated tomatoes and lettuce leaves were treated with either chilled deionized water or NEO water, with or without 20 kHz ultrasound (130 W and 210 W). Tomatoes were treated for 1, 3 and 5 min while lettuce was treated for 5, 10 and 15 min. Ultrasound significantly increased the oxidation-reduction potential (ORP) of NEO water (p < 0.05) but did not affect the pH and free chlorine concentration (p > 0.05). Increased washing time and higher ultrasonic power led to significantly greater reductions of both pathogens on produce items (p < 0.05). NEO water combined with 210 W ultrasonication for 15 min led to 4.4 and 4.3 log reductions of E. coli O157:H7 and S. Typhimurium on lettuce, respectively, while 210 W ultrasound for 5 min completely inactivated both pathogens on tomatoes. Both pathogens were completely inactivated in NEO water solutions, suggesting that its use presents little chance of cross-contamination.     

A novel electrochemical device as a disinfection system to maintain water quality during washing of ready to eat fresh produce

Electrolyzed water (EW) is known by its bactericidal efficacy and capability to oxidize organic matter. The present research evaluated the efficacy of recently developed electrolytic cells able to generate higher concentration of reactive oxygen species using lower power and salt concentration than conventional cells. This study tested the inactivation of Escherichia coli O157:H7, the organic matter depletion and trihalomethane (THM) generation by EW in process wash water under dynamic conditions. To achieve this, clean tap water was continuously added up to 60 min with artificial process water with high chemical oxygen demand (COD) inoculated with E. coli O157:H7, in experiments performed in a pilot plant that recirculated water through one electrolytic cell. Plate counts of E. coli O157:H7, COD, THMs, free, combined and total chlorine, pH, temperature and oxidation-reduction potential were determined. Results indicate that the novel electrolysis system combined with minimal addition of NaCl (0.05%) was able to suppress E. coli O157:H7 population build-up and decreased the COD accumulation in the process wash water. THM levels in the water were relatively high but its concentration in the washed product was marginal. Highly effective electrolysis has been proven to reduce the occurrence of foodborne diseases associated to cross-contamination in produce washers without having an accumulation of THMs in the washed product.     

Removal of Listeria monocytogenes, Staphylococcus aureus and Escherichia coli O157:H7 biofilms on stainless steel using scallop shell powder

Biofilms on steel surfaces containing Listeria monocytogenes, Staphylococcus aureus and Escherichia coli O157:H7 continue to threaten dairy and meat processors. In this study, the ability of scallop shell powder (SSP) to remove biofilms formed by these three pathogens on stainless steel plates was examined. Whey powder solution (WPS) and bench wash water (BWW) provided by dairy and meat factories, respectively, were inoculated with L. monocytogenes, S. aureus or E. coli O157:H7 (9 log₁₀ CFU/ml). Stainless steel plates (10 cm²) were placed in the inoculated fluids and incubated at 20 °C at 48 h to form biofilms. After drying and washing in sterile water, the plates were treated with 0.0, 0.25, or 0.50% (w/v) SSP slurries for 1, 5, or 10 min and then quantitatively examined for the three pathogens. Both 0.25 and 0.50% SSP reduced L. monocytogenes on the plates by 4 log CFU/cm² with a 1 min exposure to 0.50% SSP decreasing S. aureus by 5 logs CFU/cm². After 1 min in 0.25 and 0.50% SSP, E. coli O157:H7 populations in WPS and BWW biofilms decreased 4 and 6 log CFU/cm² and 3 and 5 log CFU/cm², respectively. Increasing the concentration of SSP led to significantly increased efficacy against the tested pathogens (P < 0.05). In conclusion, this study showed that SSP slurries could significantly reduce the numbers of L. monocytogenes, S. aureus and E. coli O157:H7 in biofilms on stainless steel surfaces.     

Effects of water hardness and pH on efficacy of chlorine-based sanitizers for inactivating Escherichia coli O157:H7 and Listeria monocytogenes

The effects of hardness and pH of water used to prepare electrolyzed oxidizing (EO) water and bleach solutions on the bactericidal activity of sanitizer prepared from the water were examined. EO water and bleach solutions were prepared with hard water of 0, 50, 100, and 200 mg/l as CaCO₃ at pH 5, 6, 7, and 8. Increased water hardness tended to increase free chlorine and oxidation–reduction potential (ORP) and decrease pH of EO water. Chlorine levels also increased with water pH. Water hardness and pH only had minor effect on the pH of bleach solutions. Increasing hardness to 50 mg/l increased antimicrobial effect of EO water against Escherichia coli O157:H7, but reduced when water hardness further increased to 100 mg/l or higher. Water pH had no effect on EO water produced against E. coli O157:H7. Water hardness had no significant effect on bactericidal activity of EO water against Listeria monocytogenes but elevated water pH decreased bactericidal activity of EO water produced against L. monocytogenes. Bleach solution prepared using hard water at 200 mg/l or at pH 7 or higher had significant lower efficacy in inactivating E. coli O157:H7, but had no effect on the inactivation of L. monocytogenes. Results indicate that increasing the hardness or pH of water used to prepare EO water or bleach solutions will decrease the bactericidal activity of sanitizers prepared from the water.     

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.     

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.     

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.     

Modeling the inactivation of Escherichia coli O157:H7 and Uropathogenic E. coli in ground beef by high pressure processing and citral

Escherichia coli O157:H7 are a well-known intestinal foodborne pathogen which were responsible for numerous foodborne illness outbreaks. Uropathogenic E. coli (UPEC) are common contaminants in meat and poultry, and may cause urinary tract infections after colonizing the gastrointestinal tract followed by accidental transfer of contaminated feces from the anus to the urethra. High pressure processing (HPP) has been demonstrated an effective means in reducing pathogenic E. coli levels in meat and poultry. Citral, an antimicrobial, also demonstrated some inactivation effect on pathogenic E. coli in ground beef (ca. 0.5–1.0 log CFU/g reduction at 1% w/w without HPP). With HPP alone, to achieve 5 log CFU/g reduction required a 500 MPa level and 15 min (for both O157:H7 and UPEC). However, 1% of citral addition may lower the pressure requirement to 380 MPa and 15 min which could reduce the food quality damage. To effectively inactivate E. coli O157:H7 and UPEC in meat, high pressure processing (HPP) in combination with the antimicrobial citral was studied. Ground beef inoculated with E. coli O157:H7 or UPEC were treated at different HPP conditions (250–350 MPa; 10–20 min), and citral (0.75–1.25%, w/w) following a central composite experimental design. Quadratic linear regression equations were developed to describe and predict the reductions of E. coli O157:H7 (R² = 0.93, p < 0.001) and UPEC (R² = 0.92, p < 0.001). Dimensionless nonlinear models consisting of three impact factors were also developed and compared with the linear models. These models were experimentally validated. Citral enhanced the inactivation of pathogenic E. coli, increasing the effectiveness of the HPP process. The models may assist the food industry and regulatory agencies in risk assessment of E. coli O157:H7 and UPEC on ground meats.     

Combined treatment with chlorine dioxide gas,fumaric acid,and ultraviolet-C light for inactivating Escherichia coli O157:H7 and Listeria monocytogenes inoculated on plums

Combined non-thermal treatment with chlorine dioxide (ClO₂) gas, ultraviolet-C (UV-C) light, and fumaric acid was performed to inactivate Listeria monocytogenes and Escherichia coli O157:H7 inoculated on plums. Plums were treated with ClO₂ gas (15 and 30 ppmv), fumaric acid (0.1, 0.3, and 0.5%), and by UV-C irradiation (3, 5, and 10 kJ/m²). The single treatments with 15 or 30 ppmv ClO₂ gas, 0.5% fumaric acid, and 10 kJ/m² UV-C decreased the population of L. monocytogenes by 1.78, 2.00, 1.65, and 1.62 log CFU/g, respectively, and the population of E. coli O157:H7 by 1.73, 1.81, 1.34, and 2.07 log CFU/g, respectively. In addition, combined treatments reduced the populations of the pathogenic bacteria more than each treatment alone. In particular, the combined treatment with ClO₂ gas (30 ppmv) for 20 min, fumaric acid (0.5%), and UV-C (10 kJ/m²) decreased the populations of L. monocytogenes and E. coli O157:H7 by 6.26 and 5.48 log CFU/g, respectively. These results suggest that combined treatment with ClO₂ gas, UV-C light, and fumaric acid may be a useful hurdle technology to enhance the microbiological safety of plums.     

Efficacy of near neutral and alkaline pH electrolyzed oxidizing waters to control Escherichia coli O157:H7 and Salmonella Typhimurium DT 104 from beef hides

In this study efficacy of near neutral and alkaline pH electrolyzed oxidizing waters to reduce aerobic plate counts (AC) and Enterobacteriaceae (EC) from uninoculated fresh cattle hides and Escherichia coli O157:H7 and Salmonella Typhimurium DT 104 from inoculated hides were determined. Fresh hides were cut in to 15 by 20 cm pieces and subjected to a total of eight different treatment solutions; near neutral pH EO water (NEW-pH 6.5 at room temperature, 150 mg/L available chlorine), alkaline pH EO water (AEO-pH 11.6 at room temperature), hot alkaline pH EO water at 43 °C (HAEO-pH 11.60), alkaline pH EO water spray followed by 150 mg/L available chlorine containing near neutral pH EO water spray (A-NEW-both at room temperature), Blitz™ (PAA, pH 3.02 at room temperature), 5% lactic acid (LA, pH 2.04 at room temperature), deionized water (W) and no treatment (Control). For each treatment, 60 ml treatment solution was sprayed on hide using a hand held sprayer. Similar treatment protocol was employed to treat hide pieces inoculated with E. coli O157:H7 and S. Typhimurium DT 104. Five percent lactic acid spray treatment was found to be the most effective treatment and achieved 2.77, 2.74, 2.75 and 2.98 log CFU/cm² of AC, EC, E. coli O157:H7 and S. Typhimurium DT 104 reductions, respectively. All EO water treatments were equally effective in reducing all target microorganisms, except E. coli O157:H7. HAEO and A-NEW treatments yielded significant reduction of E. coli O157:H7 compared to other EO water treatments. These results indicate that various EO water treatments could become viable options to reduce pathogens on hide during slaughter.     

The influence of acid stress on the growth of Listeria monocytogenes and Escherichia coli O157:H7 on cooked ham

Acid solutions are increasingly being used for decontaminating meat surfaces. On the surfaces of acid-treated meat, the population of microorganism is reduced due to the low pH of acids, and the subsequent growth of the microorganism is reduced due to the residual acids on meat surfaces. Microbial cells on meat surfaces subjected to acid treatments may cross-contaminate untreated meat surfaces, e.g., microorganisms on the surfaces of acid-treated cooked ham cross contaminate the untreated surfaces during slicing. The objective of this study was to examine this scenario in determining the subsequent growth of acid-treated Listeria monocytogenes and Escherichia coli O157:H7 on the surfaces of untreated meat. Cells of multiple-strain L. monocytogenes or E. coli O157:H7 were exposed to HCl solutions of pH 3, 4, or 5 and deionized water at room temperature for 24 h. The acid or deionized water-treated cells were inoculated separately onto cooked ham. Samples inoculated with L. monocytogenes were stored at 4 and 8 °C and samples inoculated with E. coli O157:H7 were stored at 10 and 12 °C. Populations of the pathogens on ham were enumerated during storage, and the lag phase durations (LPD, h) and growth rates (GR, log CFU/h) of the pathogens were determined. The populations of L. monocytogenes and E. coli O157:H7 in pH 5, 4, and 3 solutions were 1.2–3.1 and 0.6–2.4 log CFU/ml, respectively, lower than those in deionized water, indicating an increased acid stress on both microorganisms at lower pHs. L. monocytogenes subjected to pH 3 and pH 4 stresses and E. coli O157:H7 subjected to pH 3 stress exhibited significantly (p < 0.05) extended LPDs and reduced GRs on cooked ham. The growth of L. monocytogenes on ham was more readily reduced by acid stress than that of E. coli O157:H7. This study showed that acid treatments reduced the viability of L. monocytogenes and E. coli O157:H7and the acid stress reduced their subsequent growth ability on untreated ham. Therefore, cross-contamination of L. monocytogenes or E. coli O157:H7 cells from acid-treated meat surfaces onto untreated meat surfaces may not impose increased risk to the product.     

Inactivation of Shiga toxin-producing and nonpathogenic Escherichia coli in non-intact steaks cooked in a radio frequency oven

This study evaluated the thermal inactivation of Shiga toxin-producing Escherichia coli (O157:H7, O26:H11 and O111) (STEC) and non-pathogenic E. coli in non-intact beefsteaks (NIBS) cooked by radio frequency (RF). Blade tenderized steaks were inoculated with nalidixic (Nal) acid resistant E. coli strains, vacuum packaged in thermal pouches and cooked using pre-determined cooking times to 60 °C (rare) or 65 °C (medium-rare) inside a RF oven. Log reduction ranged from 0.99, 3.08, 2.85 and 5.0 for O157:H7, O26:H11, O111 and non-pathogenic E. coli respectively at 60 °C and a 5.0 log reduction at 65 °C for all strains. Non-pathogenic E. coli strains selected for the present study did not behave similar to the pathogenic strains being significantly more sensitive; therefore, they were not considered for testing at 63 °C. A second part of the study focused on the extent of thermal inactivation of STEC when NIBS were cooked to 63 °C (minimum safe cooking temperature recommended by USDA-FSIS). There was a 5.0 log reduction for E. coli O157:H7 and E. coli O111; but not for E. coli O26:H11. The results indicated that cooking steaks to 65 °C with a holding time at room temperature of 5 min before refrigeration would be enough to reduce numbers of E. coli O157:H7, O26:H11 and O111 using RF. The cooking protocol developed on the present study, has a practical relevance for the industry since the experiments were carried on a pilot-scale RF oven and also pathogens were tested under realistic processing conditions.     

Effect of combined ozone and organic acid treatment for control of Escherichia coli O157:H7 and Listeria monocytogenes on enoki mushroom

This study evaluated the effects of ozonated water (1, 3, and 5 ppm) alone with different exposure times (0.5, 1, 3, or 5 min), and combinations of 3 ppm ozone with 1% organic acids (acetic, citric, or lactic acids) during 5-min exposure for control of Escherichia coli O157:H7 and Listeria monocytogenes on enoki mushroom and to observe the regrowth of these pathogenic bacteria on treated enoki mushroom during storage for 10 days at 15 °C. Results showed that ozone treatment gave less than 1.0- and 0.5-log count reductions on E. coli O157:H7 and L. monocytogenes, respectively. Efficacy was improved with combined 3 ppm ozone and 1% citric acid treatment, resulting in 2.26- and 1.32-log count reductions, respectively. During storage at 15 °C (10 days) after combined treatment and packaging, populations of E. coli O157:H7 and L. monocytogenes increased to more than 8.0 log cfu/g, indicating that the combined treatment did not have a residual antimicrobial effect during storage. Although the storage study did not show control of these pathogens, the combined ozone–organic acid treatment was more effective than individual treatments in reducing initial population levels of these pathogens on enoki mushroom.     

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.     

Operating conditions for the electrolytic disinfection of process wash water from the fresh-cut industry contaminated with E. coli o157:H7

The effect of operating conditions (current density, recirculation flow rate and electrode doping level) on the efficacy of boron-doped diamond (BDD) electrodes to inactivate microorganisms and decrease chemical oxygen demand (COD) was studied in lettuce process wash water with a COD of 725 mg/L and inoculated with a 5-strain cocktail of Escherichia coli O157:H7. Changes in pathogen population, COD, pH, temperature, redox potential, and free and total chlorine were monitored in process wash water during treatments. Considering the specific characteristics of the washing step included in the fresh-cut processing, the disinfection of process wash water should be of fast action. A biphasic with a shoulder model was used to estimate shoulder length (Sl), log-linear inactivation rates (k_max1, k_max2), lowest population (N_f) and highest log reduction (HLR). Current density clearly influenced Sl, and k_max2; recirculation flow rate influenced Sl, k_max1, k_max2 and COD depletion; and doping level influenced N_f. No relationship was observed between inactivation parameters and chlorine concentration. Conditions including high current density (180 mA/cm²), high flow rate (750 l/h) and high doping level (8 000 μmol/mol) seems to provide a disinfection efficiency suitable to decrease the chance of bacterial cross contamination in the fresh-cut industries while saving on water consumption and decreasing the amount of wastewater effluents.     

The effect of different processing parameters on the efficacy of commercial post-harvest washing of minimally processed spinach and shredded lettuce

The effect of different processing parameters on the efficacy of commercial post-harvest biocidal washes to decrease the bacterial loading on spinach and lettuce has been evaluated. Sampling was performed at two spinach processors (Facility A & B) and a shredded lettuce producer (Facility C). Aerobic colony counts (ACC) and coliform counts were determined on samples taken at pre- and post-wash. In parallel, the heterotrophic plate count (HPC) and coliform levels in wash water was also determined. Processing parameters measured were the temperature of leafy greens (pre- and post-washing) and wash water. The sanitizer levels (peroxyacetic acid, oxidation-reduction potential), pH, conductivity and turbidity were also measured. The wash process in Facility B had a residence time of 50 s for the spinach, maintained a constant hypochlorite concentration and continuously re-charged the tanks with fresh water. In contrast, Facility A had a short residence time (15 s) did not maintain a constant sanitizer (peroxyacetic acid) concentration or re-charge tanks with fresh water. Despite the differences in processing operations there was no statistical difference between the log count reductions (LCR) obtained in ACC and coliform counts although counts were only reduced by <0.6 log cfu/g. The carriage of Escherichia coli on pre-wash spinach was 19% and 25% in Facility A and B respectively. There was a high prevalence (57% positive) of E. coli in the wash water of Facility A with none being recovered in water samples taken from Facility B. Yet, the carriage of E. coli on post-wash spinach was the same in the two facilities (7%). Lettuce harboured a lower level of both ACC and coliforms with LCR being significantly greater than spinach. In general, the LCR in ACC and coliforms could be positively correlated to bacterial counts of pre-washed leafy greens and conductivity (solids content) of the wash water. A negative correlation was found between LCR and water temperature. Interestingly, within the ranges measured the LCR was independent of the bacterial loading of the water. The results of the study confirmed the limited efficacy of biocidal washes to remove field acquired contamination. Although it is thought maintaining a low microbial loading in the wash water and maintaining sanitizer concentration is key the current study suggests high conductivity and low temperature of the wash water enhances the LCR achieved.     

Pulsed electric field inactivation of E. coli O157:H7 and non-pathogenic surrogate E. coli in strawberry juice as influenced by sodium benzoate,potassium sorbate,and citric acid

Current FDA regulations require that juice processors achieve a 5 log CFU/ml reduction of a target pathogen prior to distributing products. Whereas thermal pasteurization reduces the sensory characteristics of juice, pulsed electric field (PEF) treatments can be conducted at lower temperatures and may preserve sensory characteristics.Escherichia coli O157:H7 (ATCC 43895) and a non-pathogenic E. coli (ATCC 35218), respectively, were inoculated into single-strength strawberry juice with or without 750 ppm sodium benzoate (SB), 350 ppm potassium sorbate (PS), and 2.7% citric acid (CA). Juice was treated at outlet temperatures of 45, 50 and 55 °C at a field strength of 18.6 kV/cm for 150 μs with a laboratory-scale PEF unit. Inactivation of surrogate E. coli at 45, 50, and 55 °C were 2.86, 3.12, and 3.79 log CFU/ml, respectively, in plain juice (pH 3.4), and 2.75, 3.52, and 5.11 with the addition of benzoic and sorbic acids (pH 3.5). Inactivation of E. coli O157:H7 under the same conditions were 3.09, 4.08, and 4.71 log CFU/ml, respectively, and 2.27, 3.29, and 5.40 with antimicrobials. E. coli O157:H7 in juice with antimicrobials and 2.7% CA (pH 2.7) treated with PEF was reduced by 2.60, 4.32 and 6.95 log CFU/ml at 45, 50 and 55 °C while the surrogate E. coli decreased by 3.54, 5.69, and 7.13 log under the same conditions. When juice (pH 2.7) was held for 6 h without PEF treatment, higher numbers of E. coli 35218 (7.17 log CFU/ml) were inactivated than of acid-resistant E. coli O157:H7 (3.89 log). Slightly greater PEF inactivation of E. coli O157:H7 than of the surrogate bacterium indicates that E. coli ATCC 35218 provides a margin of safety when used as a surrogate for O157:H7 in plain strawberry juice or in juice + SB + PS at 45–50 °C, or with SB + PS and CA at 55 °C.