Inactivation of Listeria monocytogenes and Escherichia coli O157:H7 biofilms by micelle-encapsulated eugenol and carvacrol

Carvacrol and eugenol were encapsulated in micellar nonionic surfactant solutions to increase active component concentrations in the aqueous phase and used to treat two strains of Listeria monocytogenes (Scott A and 101) and two strains of Escherichia coli O157:H7 (4388 and 43895) grown as biofilms in a Centers for Disease Control and Prevention reactor. L. monocytogenes biofilms were grown in two different growth media, 1:20 TSB and Modified Welshimer's broth (MWB), while E. coli O157:H7 was grown in M9. In general, L. monocytogenes strains were more resistant to both micelle-encapsulated antimicrobials than E. coli O157:H7 strains. The two antimicrobials were equally effective against both strains of E. coli O157:H7, decreasing viable counts by 3.5 to 4.8 log CFU/cm(2) within 20 min. For both bacteria, most of the bactericidal activity took place in the first 10 min of antimicrobial exposure. Biofilm morphology and viability were assessed by the BacLight RedoxSensor CTC Vitality kit and confocal scanning laser microscopy, revealing an increasing number of dead cells when biofilms were treated with sufficiently high concentrations of carvacrol- or eugenol-loaded micelles. This study demonstrates the effectiveness of the application of surfactant-encapsulated essential oil components on two pathogen biofilm formers such as E. coli O157:H7 and L. monocytogenes grown on stainless steel coupons.     

Effects of chlorine and pH on efficacy of electrolyzed water for inactivating Escherichia coli O157:H7 and Listeria monocytogenes

The effects of chlorine and pH on the bactericidal activity of electrolyzed (EO) water were examined against Escherichia coli O157:H7 and Listeria monocytogenes. The residual chlorine concentration of EO water ranged from 0.1 to 5.0 mg/l, and the pH effect was examined at pH 3.0, 5.0, and 7.0. The bactericidal activity of EO water increased with residual chlorine concentration for both pathogens, and complete inactivation was achieved at residual chlorine levels equal to or higher than 1.0 mg/l. The results showed that both pathogens are very sensitive to chlorine, and residual chlorine level of EO water should be maintained at 1.0 mg/l or higher for practical applications. For each residual chlorine level, bactericidal activity of EO water increased with decreasing pH for both pathogens. However, with sufficient residual chlorine (greater than 2 mg/l), EO water can be applied in a pH range between 2.6 (original pH of EO water) and 7.0 while still achieving complete inactivation of E. coli O157:H7 and L. monocytogenes.     

Efficacy of neutral electrolyzed water to inactivate Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, and Staphylococcus aureus on plastic and wooden kitchen cutting boards

This study evaluated the efficacy of neutral electrolyzed water (NEW; 64.1 mg/liter of active chlorine) to reduce populations of Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Listeria monocytogenes on plastic and wooden kitchen cutting boards. Its effectiveness was compared with that of a sodium hypochlorite solution (NaClO; 62.3 mg/liter of active chlorine). Inoculated portions of cutting boards were rinsed in either NEW or NaClO solutions, or deionized water (control). Plastic boards were rinsed for 1 min and wooden boards for 1 and 5 min. After each treatment, the surviving population of each strain was determined on the surface and in the soaking water. No significant difference (P > or = 0.05) was found between the final populations of each strain with regard to the treatment solutions (NEW or NaClO). However, a significant difference (P < or = 0.05) was revealed between surface materials after 1 min of washing. Whereas in plastic boards the initial bacterial populations were reduced by 5 log CFU/50 cm2, in wooden cutting boards they underwent a reduction of <3 log CFU/50 cm2. A 5-min exposure time yielded reductions of about 4 log CFU/50 cm2. The surviving populations of all bacteria in NEW and NaCIO washing solutions were <1 log CFU/ml after soaking both surfaces. This study revealed that NEW treatment is an effective method for reducing microbial contamination on plastic and wooden cutting boards. NEW efficacy was comparable to that of NaCIO, with the advantage of having a larger storage time.     

Treatment of Escherichia coli O157:H7 inoculated alfalfa seeds and sprouts with electrolyzed oxidizing water

Electrolyzed oxidizing water is a relatively new concept that has been utilized in agriculture, livestock management, medical sterilization, and food sanitation. Electrolyzed oxidizing (EO) water generated by passing sodium chloride solution through an EO water generator was used to treat alfalfa seeds and sprouts inoculated with a five-strain cocktail of nalidixic acid resistant Escherichia coli O157:H7. EO water had a pH of 2.6, an oxidation-reduction potential of 1150 mV and about 50 ppm free chlorine. The percentage reduction in bacterial load was determined for reaction times of 2, 4, 8, 16, 32, and 64 min. Mechanical agitation was done while treating the seeds at different time intervals to increase the effectiveness of the treatment. Since E. coli O157:H7 was released due to soaking during treatment, the initial counts on seeds and sprouts were determined by soaking the contaminated seeds/sprouts in 0.1% peptone water for a period equivalent to treatment time. The samples were then pummeled in 0.1% peptone water and spread plated on tryptic soy agar with 5 microg/ml of nalidixic acid (TSAN). Results showed that there were reductions between 38.2% and 97.1% (0.22-1.56 log(10) CFU/g) in the bacterial load of treated seeds. The reductions for sprouts were between 91.1% and 99.8% (1.05-2.72 log(10) CFU/g). An increase in treatment time increased the percentage reduction of E. coli O157:H7. However, germination of the treated seeds reduced from 92% to 49% as amperage to make EO water and soaking time increased. EO water did not cause any visible damage to the sprouts.     

Inactivation of Escherichia coli O157: H7 and Listeria monocytogenes by PR-26, a synthetic antibacterial peptide.

This study reports the antibacterial effect of PR-26, a synthetic peptide derived from the first 26 amino acid sequence of PR-39, an antimicrobial peptide isolated from porcine neutrophils. A three-strain mixture of Escherichia coli O157:H7 or Listeria monocytogenes of approximately 10(8) CFU was inoculated to a final concentration of 10(7) CFU/ml in 1% peptone water (pH 7.0), containing 50 or 75 microg/ml of PR-26, and incubated at 37 degrees C for 0, 6, 12, and 24 h; at 24 degrees C for 0, 12, 24, and 36 h; or at 10 or 4 degrees C for 0, 24, 72, and 120 h. Control samples included 1% peptone water inoculated with each pathogen mixture but containing no PR-26. The surviving population of each pathogen at each sampling time was determined by plating on tryptic soy agar with incubation at 37 degrees C for 24 h. At 37 degrees C, PR-26 decreased E. coli O157:H7 and L. monocytogenes populations by >5.0 log CFU/ml at 12 h, with complete inactivation at 24 h. At 24 degrees C, PR-26 reduced E. coli O157:H7 and L. monocytogenes by approximately 3.5, 4.0, and 4.5 log CFU/ml at the end of 12-, 24-, and 36-h incubations, respectively. At 4 and 10 degrees C, the inhibitory effect of PR-26 on E. coli O157:H7 and L. monocytogenes was significantly lower (P < 0.05) than that at 37 and 24 degrees C: a 2- to 3-log CFU/ml reduction was observed at 120-h incubation. Results indicate that PR-26 could potentially be used as an antimicrobial agent, but applications in appropriate foods need to be validated.     

Inactivation of Escherichia coli O157:H7 and Listeria monocytogenes inoculated on raw salmon fillets by pulsed UV-light treatment

The efficacy of pulsed UV‐light to inactivate of Escherichia coli O157:H7 and Listeria monocytogenes Scott A on salmon fillets was investigated in this study by evaluating the effects of treatment times and distance from the UV strobe. The sterilization system generated 5.6 J cm^?2 per pulse at the lamp surface for an input voltage of 3800 V and three pulses per second. Skin or muscle side inoculated salmon fillet (8 cm × 1.5 cm) in a Petri dish was placed on shelf at three different distances from the UV strobe; 3, 5, and 8 cm. At each distance, the pulsed UV‐light treatment was performed for 15, 30, 45, and 60 s. For E. coli O157:H7, maximum log₁₀ reduction was 1.09 log₁₀ CFU g^?1 on muscle side at 8 cm for 60‐s treatment, whereas 0.86 log₁₀ CFU g^?1 reduction on skin at 5 cm for 30‐s treatment. For L. monocytogenes Scott A, maximum reduction was 1.02 log₁₀ CFU g^?1 at 8 cm for 60‐s treatment on skin side, whereas 0.74 log₁₀ CFU g^?1 reduction on muscle at 8 cm for 60‐s treatment. The fillet's surface temperature increased up to 100degrC within 60‐s treatment time. Therefore, some fish samples were overheated after 30 and 45 s at 3‐ and 5‐cm distances from light source, respectively, which resulted in visual colour and quality changes. Overall, this study demonstrated that about one log reduction (c. 90%) of E. coli O157:H7 or L. monocytogenes could be achieved at 60‐s treatment at 8 cm distance without affecting the quality.     

Reduction of Salmonella enterica, Escherichia coli O157:H7, and Listeria monocytogenes with electrolyzed oxidizing water on inoculated hass avocados (Persea americana var. Hass)

This study was intended to evaluate the bactericidal effect of electrolyzed oxidizing water (EOW) and chlorinated water on populations of Salmonella enterica, Escherichia coli O157:H7, and Listeria monocytogenes inoculated on avocados (Persea americana var. Hass). In the first experiment, inoculated avocados were treated with a water wash applied by spraying tap water containing 1 mg/liter free chlorine for 15 s (WW); WW treatment and then spraying sodium hypochlorite in water containing 75 mg/liter free chlorine for 15 s (Cl75); WW treatment and then spraying alkaline EOW for 30 s (AkEW) and then spraying acid EOW (AcEW) for 15 s; and spraying AkEW and then AcEW. In another experiment, the inoculated avocados were treated by spraying AkEW and then AcEW for 15, 30, 60, or 90 s. All three pathogen populations were lowered between 3.6 and 3.8 log cycles after WW treatment. The application of Cl75 did not produce any further reduction in counts, whereas AkEW and then AcEW treatment resulted in significantly lower bacterial counts for L. monocytogenes and E. coli O157:H7 but not for Salmonella. Treatments with AkEW and then AcEW produced a significant decrease in L. monocytogenes, Salmonella, and E. coli O157:H7 populations, with estimated log reductions of 3.9 to 5.2, 5.1 to 5.9, and 4.2 to 4.9 log CFU/cm2, respectively. Spraying AcEW for more than 15 s did not produce any further decrease in counts of Salmonella or E. coli O157:H7, whereas L. monocytogenes counts were significantly lower after spraying AcEW for 60 s. Applying AkEW and then AcEW for 15 or 30 s seems to be an effective alternative to reduce bacterial pathogens on avocado surfaces.     

Survival of Listeria monocytogenes and Escherichia coli O157:H7 during sauerkraut fermentation

Sauerkraut was produced from shredded cabbage, as is typical in the United States, and from whole head cabbages, which is a traditional process in parts of Eastern Europe. The sauerkraut was inoculated with five strain mixtures of Escherichia coli O157:H7 and Listeria monocytogenes, and the populations of these bacteria, as well as lactic acid bacteria, pH, and titratable acidity, were monitored over the course of fermentation. Fermentation variables were temperature (18 and 22 degrees C) and salt concentration (1.8, 2.25, and 3%). For most of the analyses, the type of cabbage processing was a significant factor, although within cabbage type, neither salt nor fermentation temperature had significant effects. The final pH of the whole-head sauerkraut was lower than the shredded sauerkraut, but the titratable acidity was significantly higher in the shredded sauerkraut. E. coli O157:H7 and L. monocytogenes persisted in the brines for most of the fermentation, although at the end of the fermentations (15 days for shredded, 28 days for whole head), neither pathogen had detectable populations. E. coli populations decreased more rapidly in the shredded sauerkraut even though the pH was higher because of the higher total acidity in the shredded sauerkraut. Acid-tolerant strains of E. coli and L. monocytogenes were isolated from both shredded and whole-head sauerkraut at different salt concentrations and temperatures after 15 days of fermentation and could be detected at 35 days in the wholehead sauerkraut.     

Effectiveness of electrolyzed acidic water in killing Escherichia coli O157:H7, Salmonella enteritidis,and Listeria monocytogenes on the surfaces of tomatoes

A study was conducted to evaluate the efficacy of electrolyzed acidic water, 200-ppm chlorine water, and sterile distilled water in killing Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes on the surfaces of spot-inoculated tomatoes. Inoculated tomatoes were sprayed with electrolyzed acidic water, 200-ppm chlorine water, and sterile distilled water (control) and rubbed by hand for 40 s. Populations of E. coli O157:H7, Salmonella, and L. monocytogenes in the rinse water and in the peptone wash solution were determined. Treatment with 200-ppm chlorine water and electrolyzed acidic water resulted in 4.87- and 7.85-log10 reductions, respectively, in Escherichia coli O157:H7 counts and 4.69- and 7.46-log10 reductions, respectively, in Salmonella counts. Treatment with 200-ppm chlorine water and electrolyzed acidic water reduced the number of L. monocytogenes by 4.76 and 7.54 log10 CFU per tomato, respectively. This study's findings suggest that electrolyzed acidic water could be useful in controlling pathogenic microorganisms on fresh produce.     

Inactivation kinetics of inoculated Escherichia coli O157:H7, Listeria monocytogenes and Salmonella enterica on strawberries by chlorine dioxide gas

Inactivation kinetics of inoculated Escherichia coli O157:H7, Listeria monocytogenes and Salmonella enterica on strawberries by chlorine dioxide gas at different concentrations (0.5, 1, 1.5, 3 and 5 mgl(-1)) for 10 min were studied. A cocktail of three strains of each targeted organism (100 microl) was spotted onto the surface of the strawberries (approximately 8-9 log ml(-1)) separately followed by air drying, and then treated with ClO(2) gas at 22 degrees C and 90-95% relative humidity. Approximately a 4.3-4.7 logCFU reduction per strawberry of all examined bacteria was achieved by treatment with 5 mgl(-1) ClO(2) for 10 min. The inactivation kinetics of E. coli O157:H7, L. monocytogenes and S. enterica were determined using first-order kinetic models to establish D-values and z-values. The D-values of E. coli, L. monocytogenes and S. enterica were 2.6+/-0.2, 2.3+/-0.2 and 2.7+/-0.7 min, respectively, at 5 mgl(-1) ClO(2). The z-values of E. coli, L. monocytogenes and S. enterica were 16.8+/-3.5, 15.8+/-3.5 and 23.3+/-3.3 mgl(-1), respectively. Furthermore, treatment with ClO(2) gas significantly (p < or = 0.05) reduced the initial microflora (mesophilic, psychrotrophic bacteria, yeasts and molds) on strawberries. Treatment with ClO(2) gas did not affect the color of strawberries and extended the shelf-life to 16 days compared to 8 days for the untreated control.     

Inactivation of Escherichia coli O157:H7, Salmonella typhimurium and Listeria monocytogenes in apple juice with gaseous ozone

This research was initiated to assess the efficacy of gaseous ozone for inactivation Escherichia coli O157:H7, Salmonella typhimurium and Listeria monocytogenes in apple juice. Juice samples with solids content of 18, 36, and 72 °Brix inoculated with a culture cocktail of three foodborne pathogens were treated with gaseous ozone at a flow rate of 3.0 L/min and an ozone generation rate of 0.10, 0.90, 3.51, and 5.57 g/h for 0.5, 1, 5, and 10 min, respectively. The inactivation kinetics of gaseous ozone on foodborne pathogens conformed to the Weibull model. The time required to achieve a 5 log reduction (t_5d) was estimated using the parameters of the Weibull model. The t_5d increased with increasing solids content of apple juice. The ozone generation rate did not impart a significant effect (p > 0.05) on t_5d. Gaseous ozone is effective at inactivating foodborne pathogens in apple juice but the efficacy is dependent on the solids content of the juice sample.     

Inactivation kinetics of inoculated Escherichia coli O157:H7, Listeria monocytogenes and Salmonella Poona on whole cantaloupe by chlorine dioxide gas

The objectives of this study were to examine inactivation kinetics of inoculated Escherichia coli O157:H7, Listeria monocytogenes and Salmonella Poona inoculated onto whole cantaloupe and treated with ClO(2) gas at different concentrations (0.5, 1.0, 1.5, 3.0 and 5.0 mg l(-1)) for different times (0, 2.0, 4.0, 6.0, 8.0 and 10.0 min). The effect of ClO(2) gas on the quality and shelf life of whole cantaloupe was also evaluated during storage at 22 degrees C for 12 days. A 100 microl inoculation of each targeted organism was spotted onto the surface (5 cm(2)) of cantaloupe rind (approximately 8-9 log CFU 5 cm(-2)) separately, air dried (60 min), and then treated with ClO(2) gas at 22 degrees C and 90-95% relative humidity for 10 min. Surviving bacterial populations on cantaloupe surfaces were determined using a membrane transferring method with a non-selective medium followed by a selective medium. The inactivation kinetics of E. coli O157:H7, L. monocytogenes and S. Poona were determined using nonlinear kinetics (Weibull model). A 3 log CFU reduction of E. coli O157:H7, L. monocytogenes and S. Poona were achieved with 5.0 mg l(-1) ClO(2) gas for 5.5, 4.2 and 1.5 min, respectively. A 5l og CFU reduction of S. Poona was achieved with 5.0 and 3.0 mg l(-1) ClO(2) gas for 6 and 8 min, respectively. A 4.6 and 4.3 log reduction was achieved after treatment with 5.0 mg l(-1) ClO(2) gas at 10 min for E. coli O157:H7 and L. monocytogenes, respectively. Treatment with 5.0 mg l(-1) ClO(2) gas significantly (p<0.05) reduced the initial microflora (mesophilic bacteria, psychrotrophic bacteria, and yeasts and molds) on cantaloupe by more than 2 log CFU cm(-2) and kept them significantly (p<0.05) lower than the untreated control during storage at 22 degrees C for 12 days. Treatment with ClO(2) gas did not significantly (p>0.05) affect the color of whole cantaloupe and extended the shelf life to 9 days compared to 3 days for the untreated control, when stored at ambient temperature (22 degrees C).     

Simultaneous detection of Escherichia coli O157:H7, Listeria monocytogenes and Salmonella strains by real-time PCR

A protocol enabling simultaneous detection of Escherichia coli O157:H7, Listeria monocytogenes and Salmonella strains was devised and evaluated using artificially contaminated fresh produce. Association of Official Analytical Chemists (AOAC)-approved polymerase chain reaction (PCR) detection methods for three human pathogens were modified to enable simultaneous and real-time detection with high throughput capability. The method includes a melting-curve analysis of PCR products, which serves as confirmatory test. The modified protocol successfully detected all three pathogens when fresh produce was washed with artificially contaminated water containing E. coli O157:H7 and S. typhimurium down to the predicted level of 1 to 10 cells/ml and L. monocytogenes at 1000 cells/ml. The ability to monitor several pathogens simultaneously will save time and increase our ability to assure food safety.     

Metabiotic effects of Fusarium spp. on Escherichia coli O157:H7 and Listeria monocytogenes on raw portioned tomatoes

The metabiotic effects of Fusarium proliferatum, F. avenaceum, and F. oxysporum on Escherichia coli O157:H7 and Listeria monocytogenes in fresh tomatoes were investigated. Tomatoes were preinoculated with the molds and incubated at 15 degrees C for 7 days; then they were inoculated separately with the pathogens, packaged in air and modified atmosphere (5% O2, 30% CO2, and 65% N2), and stored at 4, 8, and 12 degrees C for 9 days. The cell loads of pathogens and lactic acid bacteria and the pH were evaluated periodically. The data were modeled through some different mathematical models to assess the shoulder length, i.e., the time before the beginning of the exponential death phase, the 1-log reduction time (s), and the pathogen death time (deltastand). The preinoculation of tomatoes with the molds enhanced the survival of E. coli O157:H7 by prolonging shoulder length and 8 parameters; this effect, however, was not observed for L. monocytogenes. pH values did not undergo significant changes within the storage time, and the lactic acid bacteria increased from 5 to 7 log CFU/g, without significant differences among the storage temperatures or the packaging atmospheres. The results of this research showed that the use of fresh tomatoes colonized by fusaria (even if the contamination is not visible) could increase significantly the risk of outbreaks due to some pathogens that could be on the surface of fruits and vegetables as a result of cross-contamination at home or incorrect postharvest operations.     

Survival of Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella in juice concentrates

The survival of Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella was studied in apple, orange, pineapple, and white grape juice concentrates and banana puree. Pouches of juice concentrate or puree were inoculated with pathogens at a level > or = 10(3) CFU/g and stored at -23 degrees C (-10 degrees F). Pathogen survival was monitored at 6 and 24 h, once a week for four consecutive weeks, and biweekly thereafter until 12 weeks. When pathogens were not detectable by direct plating, samples were enriched in universal preenrichment broth for 72 h and plated on selective media. Results showed that E. coli O157:H7, L. monocytogenes, and Salmonella were recoverable from all five concentrates through 12 weeks of storage at -23 degrees C.     

Antimicrobial efficacy of eugenol microemulsions in milk against Listeria monocytogenes and Escherichia coli O157:H7

The antimicrobial activity of eugenol microemulsions (eugenol encapsulated in surfactant micelles) in ultrahigh-temperature pasteurized milk containing different percentages of milk fat (0, 2, and 4%) was investigated. Antimicrobial microemulsions were prepared from a 5% (wt) aqueous surfactant solution (Surfynol 485W) with 0.5% (wt) eugenol. Two strains each of Listeria monocytogenes and Escherichia coli O157:H7 previously shown to be the least and most resistant to the microemulsion in microbiological media were used to inoculate sterile milk (10(4) CFU/ml). Samples were withdrawn and plated at 0, 1, 3, 6, 12, and 24 h for enumeration. Microemulsions completely prevented growth of L. monocytogenes for up to 48 h in skim milk and reduced both strains of E. coli O157:H7 to less than detectable levels in less than 1 h. Similarly, in 2% fat milk, eugenol-Surfynol combinations reduced both strains of E. coli O157:H7 to less than detectable levels in less than 1 h but only increased the lag phase of both strains of L. monocytogenes. In full-fat milk (4% fat), microemulsions inhibited growth of the least resistant strains of L. monocytogenes and E. coli but were ineffective against the two resistant strains. Unencapsulated eugenol was slightly more or as inhibitory as microemulsions against target pathogens. Results were attributed to diffusional mass transport of antimicrobials from microemulsions to the macroemulsion (milk). Results suggest that food composition, especially fat level, may affect the efficiency of targeting of foodborne pathogens with surfactant-encapsulated antimicrobials.     

Effect of acidified sodium chlorite, chlorine, and acidic electrolyzed water on Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes inoculated onto leafy greens

Recent foodborne outbreaks implicating spinach and lettuce have increased consumer concerns regarding the safety of fresh produce. While the most common commercial antimicrobial intervention for fresh produce is wash water containing 50 to 200 ppm chlorine, this study compares the effectiveness of acidified sodium chlorite, chlorine, and acidic electrolyzed water for inactivating Escherichia coli O157:H7, Salmonella, and Listeria monocytogenes inoculated onto leafy greens. Fresh mixed greens were left uninoculated or inoculated with approximately 6 log CFU/g of E. coli O157:H7, Salmonella, and L. monocytogenes and treated by immersion for 60 or 90 s in different wash solutions (1:150, wt/vol), including 50 ppm of chlorine solution acidified to pH 6.5, acidic electrolyzed water (pH 2.1 +/- 0.2, oxygen reduction potential of 1,100 mV, 30 to 35 ppm of free chlorine), and acidified sodium chlorite (1,200 ppm, pH 2.5). Samples were neutralized and homogenized. Bacterial survival was determined by standard spread plating on selective media. Each test case (organism x treatment x time) was replicated twice with five samples per replicate. There was no difference (P > or = 0.05) in the time of immersion on the antimicrobial effectiveness of the treatments. Furthermore, there was no difference (P > or = 0.05) in survival of the three organisms regardless of treatment or time. Acidified sodium chlorite, resulted in reductions in populations of 3 to 3.8 log CFU/g and was more effective than chlorinated water (2.1 to 2.8 log CFU/g reduction). These results provide the produce industry with important information to assist in selection of effective antimicrobial strategies.     

Inactivation of Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella in cranberry, lemon, and lime juice concentrates

The production of thermally concentrated fruit juices uses temperatures high enough to achieve at least a 5-log reduction of pathogenic bacteria that can occur in raw juice. However, the transportation and storage of concentrates at low temperatures prior to final packaging is a common practice in the juice industry and introduces a potential risk for postconcentration contamination with pathogenic bacteria. The present study was undertaken to evaluate the likelihood of Escherichia coli O157:H7, Listeria monocytogenes and Salmonella surviving in cranberry, lemon, and lime juice concentrates at or above temperatures commonly used for transportation or storage of these concentrates. This study demonstrates that cranberry, lemon, and lime juice concentrates possess intrinsic antimicrobial properties that will eliminate these bacterial pathogens in the event of postconcentration recontamination. Bacterial inactivation was demonstrated under all conditions; at least 5-log Salmonella inactivation was consistently demonstrated at -23 degrees C (-10 degrees F), at least 5-log E. coli O157:H7 inactivation was consistently demonstrated at -11 degrees C (12 degrees F), and at least 5-log L. monocytogenes inactivation was consistently demonstrated at 0 degrees C (32 degrees F).     

Survival and growth of Listeria monocytogenes and enterohemorrhagic Escherichia coli O157:H7 in minimally processed artichokes

The ability of Listeria monocytogenes and Escherichia coli O157:H7 inoculated by immersion (at 4.6 and 5.5 log CFU/ g, respectively) to survive on artichokes during various stages of preparation was determined. Peeling, cutting, and disinfecting operations (immersion in 50 ppm of a free chlorine solution at 4 degrees C for 5 min) reduced populations of L. monocytogenes and E. coli O157:H7 by only 1.6 and 0.8 log units, respectively. An organic acid rinse (0.02% citric acid and 0.2% ascorbic acid) was more effective than a tap water rinse in removing these pathogens. Given the possibility of both pathogens being present on artichokes at the packaging stage, their behavior during the storage of minimally processed artichokes was investigated. For this purpose, batches of artichokes inoculated with L. monocytogenes or E. coli O157:H7 (at 5.5 and 5.2 log CFU/g, respectively) were packaged in P-Plus film bags and stored at 4 degrees C for 16 days. During this period, the equilibrium atmosphere composition and natural background microflora (mesophiles, psychrotrophs, anaerobes, and fecal coliforms) were also analyzed. For the two studied pathogens, the inoculum did not have any effect on the final atmospheric composition (10% O2, 13% CO2) or on the survival of the natural background microflora of the artichokes. L. monocytogenes was able to survive during the entire storage period in the inoculated batches, while the E. coli O157:H7 level increased by 1.5 log units in the inoculated batch during the storage period. The modified atmosphere was unable to control the behavior of either pathogen.     

Inhibition and inactivation of Listeria monocytogenes and Escherichia coli O157:H7 colony biofilms by micellar-encapsulated eugenol and carvacrol

The antimicrobial efficacy of carvacrol and eugenol, two essential oil compounds, encapsulated in a micellar nonionic surfactant solution on four strains of Listeria monocytogenes (Scott A, 101, 108, and 310) and four strains of Escherichia coli O157:H7 (H1730, E0019, F4546, and 932) growing as colony biofilms was investigated. Carvacrol and eugenol were encapsulated in Surfynol 485W at concentrations ranging from 0.3 to 0.9% (wt/wt) at a surfactant concentration of 5% (wt/wt). Colony biofilms were grown on polycarbonate membranes resting on agar plates containing antimicrobial formulations. Cells were enumerated after 0, 3, 6, 9, 24, 48, and 72 h of incubation. Colony biofilms of all E. coli O157:H7 strains were more sensitive to both antimicrobial systems than L. monocytogenes strains. Surface-grown E. coli O157:H7 viable cell numbers decreased below detectable levels after exposure to encapsulated essential oil compounds for > 3 h at all tested concentrations, except for E. coli O157:H7 F4546, which grew slowly in the presence of < 0.5% (wt/wt) eugenol. L. monocytogenes Scott A and 101 were more resistant to eugenol than carvacrol at sublethal concentrations (< 0.5% [wt/wt]). Carvacrol was effective at any concentration against L. monocytogenes 108, whereas concentrations of > 0.5% (wt/wt) eugenol were required for inactivation. L. monocytogenes 310 was equally sensitive to both essential oil compounds. Results suggest that surfactant-encapsulated generally recognized as safe essential oil compounds may offer a new means to control the growth of food pathogens such as E. coli O157:H7 and L. monocytogenes on food contact surfaces.