Inactivation of Escherichia coli O1 57:H7, Listeria monocytogenes, and Lactobacillus leichmannii by combinations of ozone and pulsed electric field

Pulsed electric field (PEF) and ozone technologies are nonthermal processing methods with potential applications in the food industry. This research was performed to explore the potential synergy between ozone and PEF treatments against selected foodborne bacteria. Cells of Lactobacillus leichmannii ATCC 4797, Escherichia coli O157:H7 ATCC 35150, and Listeria monocytogenes Scott A were suspended in 0.1% NaCl and treated with ozone, PEF, and ozone plus PEE Cells were treated with 0.25 to 1.00 microg of ozone per ml of cell suspension, PEF at 10 to 30 kV/cm, and selected combinations of ozone and PEF. Synergy between ozone and PEF varied with the treatment level and the bacterium treated. L. leichmannii treated with PEF (20 kV/cm) after exposure to 0.75 and 1.00 microg/ml of ozone was inactivated by 7.1 and 7.2 log10 CFU/ml, respectively; however, ozone at 0.75 and 1.00 microg/ml and PEF at 20 kV/cm inactivated 2.2, 3.6, and 1.3 log10 CFU/ml, respectively. Similarly, ozone at 0.5 and 0.75 microg/ml inactivated 0.5 and 1.8 log10 CFU/ml of E. coli, PEF at 15 kV/cm inactivated 1.8 log10 CFU/ml, and ozone at 0.5 and 0.75 microg/ml followed by PEF (15 kV/cm) inactivated 2.9 and 3.6 log10 CFU/ml, respectively. Populations of L. monocytogenes decreased 0.1, 0.5, 3.0, 3.9, and 0.8 log10 CFU/ml when treated with 0.25, 0.5, 0.75, and 1.0 microg/ml of ozone and PEF (15 kV/cm), respectively; however, when the bacterium was treated with 15 kV/cm, after exposure to 0.25, 0.5, and 0.75 microg/ml of ozone, 1.7, 2.0, and 3.9 log10 CFU/ml were killed, respectively. In conclusion, exposure of L. leichmannii, E. coli, and L. monocytogenes to ozone followed by the PEF treatment showed a synergistic bactericidal effect. This synergy was most apparent with mild doses of ozone against L. leichmannii.     

The influence of process parameters for the inactivation of Listeria monocytogenes by pulsed electric fields

The influence of the electric field strength, the treatment time, the total specific energy and the conductivity of the treatment medium on the Listeria monocytogenes inactivation by pulsed electric fields (PEF) has been investigated. L. monocytogenes inactivation increased with the field strength, treatment time and specific energy. A maximum inactivation of 4.77 log(10) cycles was observed after a treatment of 28 kV/cm, 2000 micros and 3490 kJ/kg. The lethal effect of PEF treatments on L. monocytogenes was not influenced by the conductivity of the treatment medium in a range of 2, 3 and 4 mS/cm when the total specific energy was used as a PEF control parameter. A mathematical model based on the Weibull distribution was fitted to the experimental data when the field strength (15-28 kV/cm), treatment time (0-2000 micros) and specific energy (0-3490 kJ/kg) were used as PEF control parameters. A linear relationship was obtained between the log(10) of the scale factor (b) and the electric field strength when the treatment time and the total specific energy were used to control the process. The total specific energy, in addition to the electric field strength and the treatment time, should be reported in order to evaluate the microbial inactivation by PEF.     

Modelling inactivation of Listeria monocytogenes by pulsed electric fields in media of different pH

A study of the effect of square-wave pulsed electric fields (PEF) on the inactivation of Listeria monocytogenes in McIlvaine buffer of different pH (3.5-7.0) was conducted. L. monocytoges was more PEF sensitive at higher electric field strengths (E) and in media of low pH. A treatment at 28 kV/cm for 400 mus that inactivated 1.5, 2.3 and 3.0 Log10 cycles at pH 7.0, 6.5 and 5.0 respectively destroyed almost 6.0 Log10 cycles at pH 3.5. The general shape of survival curves of L. monocytogenes PEF treated at different pH was convex/concave upwards. A mathematical model based on the Weibull distribution accurately described these survival curves. At each pH, the shape parameter (n value) did not depend on E. The relationship between n value of the Weibull model and the pH of the treatment medium was described by the Gompertz equation. A multiple linear regression model using three predictor variables (E, E2, pH2) related the Log10 of the scale paramenter (b value) of the Weibull model with E and pH of the treatment medium. A tertiary model developed using McIlvaine buffer as treatment medium predicted satisfactorily the inactivation of L. monocytogenes in apple juice.     

Behavior of Listeria monocytogenes inoculated on cantaloupe surfaces and efficacy of washing treatments to reduce transfer from rind to fresh-cut pieces

Attachment and survival of Listeria monocytogenes on external surfaces (rind) of inoculated cantaloupe, resistance of the surviving bacteria to chlorine or hydrogen peroxide treatments, transfer of the pathogen from unsanitized and sanitized rinds to fresh-cut tissues during cutting and growth, and survival of L. monocytogenes on fresh-cut pieces of cantaloupe were investigated. Surface treatment with 70% ethanol to reduce the native microflora on treated melon, followed by immersion in a four-strain cocktail of L monocytogenes (10(8) CFU/ml) for 10 min, deposited 4.2 log10 CFU/cm2 and 3.5 log10 CFU/cm2 of L monocytogenes on treated and untreated cantaloupe rinds, respectively. L. monocytogenes survived on the treated or untreated cantaloupe rinds for up to 15 days during storage at 4 and 20 degrees C, but populations declined by approximately 1 to 2 log10 CFU/cm2. Fresh-cut pieces prepared from inoculated whole cantaloupes stored at 4 degrees C for 24 h after inoculation were positive for L. monocytogenes. Washing inoculated whole cantaloupes in solutions containing 1,000 ppm of chlorine or 5% hydrogen peroxide for 2 min at 1 to 15 days of storage at 4 degrees C after inoculation resulted in a 2.0- to 3.5-log reduction in L. monocytogenes on the melon surface. Fresh-cut pieces prepared from the sanitized melons were negative for L. monocytogenes. After direct inoculation onto fresh-cut pieces, L. monocytogenes survived, but did not grow, during 15 days of storage at 4 degrees C. Growth was evident by 4 h of storage at 8 and 20 degrees C. It is concluded that sanitizing with chlorine or hydrogen peroxide has the potential to reduce or eliminate the transfer of L. monocytogenes on melon surfaces to fresh-cut pieces during cutting.     

Effect of vanillin on the fate of Listeria monocytogenes and Escherichia coli O157:H7 in a model apple juice medium and in apple juice

The effects of vanillin on the fates of Listeria monocytogenes and Escherichia coli O157:H7 at pH values between 3.5 and 4.5 were verified in a model apple juice (MAJ) medium and in apple juice incubated at 4 or 15 degrees C. Viable E. coli O157:H7 cells were recovered from MAJ for up to 10 days, but L. monocytogenes did not survive at pH 3.5. Supplementation with 40 mm vanillin exerted a lethal effect that was species, concentration, pH and temperature dependant. E. coli O157:H7 was more sensitive to vanillin than L. monocytogenes, and viable cells could not be recovered after 2 days incubation at either temperature. L. monocytogenes and E. coli O157:H7 were inoculated (10(5) cfu/ml) in pH adjusted (pH 4.00) or unadjusted (pH 3.42) juice from Granny Smith apples that was supplemented with 40 mm vanillin. Neither species were recovered after 3 days incubation at 4 or 15 degrees C. These findings indicate that vanillin could be useful as a preservative for minimally processed apple products.     

Plasmin Inactivation with Pulsed Electric Fields

Plasmin (fibrinolysin E.C.3.4.21.7), an indigenous enzyme in bovine milk, added to simulated milk ultrafiltrate (SMUF) at 100 μg/mL (pH 6.11 and ionic strength 0.056 M) was treated at 10°C and 15°C with pulsed electric fields (HVPEF) of 15, 30 and 45 kV/cm and number of pulses 10, 20, 30, 40 and 50. The plasmin activity measured using a commercial assay, was reduced 90% after 50 pulses at both 30 and 45 kV/cm and at a processing temperature of 15°C. Similar inactivation was obtained when plasmin (100 μg/mL) in SMUF was heated at 40°C for 15 min. Inactivation of the enzyme depended on the number of pulses applied during treatment, intensity of the applied field, and processing temperature.     

Inhibition, survival and growth of Listeria monocytogenes on poultry as influenced by buffered lactic acid treatment and modified atmosphere packaging.

The effect of the treatment with various concentrations (2%, 5% and 10% w/v) of lactic acid/sodium lactate buffer (pH 3.0), modified atmosphere (MAP) packaging (90% CO2 and 10% O2) and 10% (w/v) lactic acid/sodium lactate buffer (pH 3.0) combined with MAP on Listeria monocytogenes Z7 serotype 1 and on the shelf life of chicken legs stored at 6 degrees C was investigated. The initial contamination level of L. monocytogenes on the chicken legs surface was 8.3 x 10(2) cfu/cm2 of skin. After 2 days of storage at 6 degrees C the number of L. monocytogenes on legs treated with 2%, 5%, 10% lactic acid/sodium lactate buffer (pH 3.0) and 10% lactic acid/sodium lactate buffer (pH 3.0) combined with MAP was significantly lower than the initial number of L. monocytogenes. Later, growth of L. monocytogenes was observed. After 13 days of storage at 6 degrees C the number of L. monocytogenes on legs treated with 10% lactic acid/sodium lactate buffer (pH 3.0) combined with MAP was still similar to the initial number. Legs treated with 2%, 5%, 10% lactic acid/sodium lactate buffer (pH 3.0), MAP and 10% lactic acid/sodium lactate buffer (pH 3.0) combined with MAP, have a shelf life at 6 degrees C of respectively 8, 9, 10, 13 and 17 days. This means a prolongation of 2, 3, 4, 7 and 11 days, respectively for storage at 6 degrees C. The antimicrobial effect of lactic acid buffer systems (pH 3.0) increased with increasing concentrations of lactic acid in the buffered system. The best results were obtained by the combined use of 10% acid/sodium lactate buffer (pH 3.0) and MAP.     

Screening for Listeria monocytogenes surrogate strains applicable to food processing by ultrahigh pressure and pulsed electric field

Ultrahigh pressure (UHP) and pulsed electric field (PEF) are emerging processing technologies developed to enhance the safety while maintaining the fresh-like quality of food. For each food and process combination, a pathogen of concern (i.e., target pathogen) must be determined, and a low-risk microorganism that serves as the pathogen surrogate for process validation must be identified. The objective of this study was to identify a surrogate for Listeria monocytogenes for UHP and PEF process validation. Potential surrogates tested include four Lactobacillus spp., a Pediococcus sp., and a Listeria innocua strain. These were compared with nine L. monocytogenes strains, with regard to sensitivity to UHP and PEF processing. For UHP treatment, the strains were suspended in citrate-phosphate buffer (pH 7.0 or 4.5), sweet whey, or acidified whey and pressure processed at 500 MPa for 1 min. For PEF treatment, the strains were suspended in NaCl solution, acid whey, or sweet whey and processed at 25 kV/cm. The lethality of UHP or PEF treatment varied considerably, depending on medium types and pH and the treated strain. Treating the tested microorganisms with UHP inactivated 0.3 to 6.9 log CFU/ml for L. monocytogenes strains and 0.0 to 4.7 log CFU/ml for the potential surrogates. When PEF was employed, populations of tested microorganisms decreased < 1.0 to 5.3 log CFU/ml. L. monocytogenes V7 and OSY-8578 were among the most resistant strains to UHP and PEF treatments, and thus are candidate target strains. Lactobacillus plantarum ATCC 8014 demonstrated similar or greater resistance compared with the target organisms; therefore, the bacterium is proposed as a surrogate of L. monocytogenes for both processes under the conditions specified in the food matrices tested in this study.     

Transfer of Listeria monocytogenes during mechanical slicing of turkey breast, bologna, and salami

A commercial delicatessen slicer was used as the vector for sequential quantitative transfer of Listeria monocytogenes (i) from an inoculated slicer blade (approximately 10(8), 10(5), or 10(3) CFU per blade) to 30 slices of uninoculated delicatessen turkey, bologna, and salami, (ii) from inoculated product (approximately 10(8) CFU/cm2) to the slicer, and (iii) from inoculated product (10(8), 10(5), or 10(3) CFU/cm2) to 30 slices of uninoculated product via the slicer blade. Cutting force and product composition also were assessed for their impact on L. monocytogenes transfer. Five product contact areas on the slicer, which were identified from residue of product bathed in Glow-Germ, were also sampled using a 1-ply composite tissue technique after inoculated product had been sliced. After being sliced with inoculated blades, each product slice was surface or pour plated on modified Oxford agar and/ or enriched in University of Vermont medium. Greater transfer (P < 0.05) occurred from inoculated turkey (10(8) CFU/cm2) to the five slicer contact areas from an application force of 4.5 kg as compared with 0 kg. On uninoculated product sliced with blades inoculated at 10(8) CFU per blade, L. monocytogenes populations decreased logarithmically to 10(2) CFU per slice after 30 slices. Findings for the inoculated slicer blade and product (10(5) CFU per blade or cm2) were similar; L. monocytogenes concentrations were 102 CFU per slice after 5 slices and enriched samples were generally negative for L. monocytogenes after 27 slices. For uninoculated product sliced with blades inoculated at 10(3) CFU per blade, the first 5 slices typically produced L. monocytogenes at approximately 10 CFU per slice by direct plating, and enrichments were negative for L. monocytogenes after 15 slices. The higher fat and lower moisture content of salami compared with turkey and bologna resulted in a visible fat layer on the blade that likely prolonged L. monocytogenes transfer. As a result of cross-contamination, those delicatessen-sliced meats that allow growth of L. monocytogenes during prolonged refrigerated storage likely pose an increased public health risk for certain consumers.     

Biofilm formation by acid-adapted and nonadapted Listeria monocytogenes in fresh beef decontamination washings and its subsequent inactivation with sanitizers

The antimicrobial effects of sodium hypochlorite (SH, 200 ppm, at an adjusted pH of 6.80 +/- 0.20 and at an unadjusted pH of 10.35 +/- 0.25), quaternary ammonium compound (pH 10.20 +/- 0.12, 200 ppm), and peroxyacetic acid (PAA, pH 3.45 +/- 0.20, 150 ppm) on previously acid-adapted or nonadapted Listeria monocytogenes inoculated (10(5) CFU/ml) into beef decontamination water washings were evaluated. The effects of the sanitizers on suspended cells (planktonic or deattached) and on cells attached to stainless steel coupons obtained from inoculated washings stored at 15 degrees C for up to 14 days were studied. Cells were exposed to sanitizers on days 2, 7, and 14. The pathogen had formed a biofilm of 5.3 log CFU/cm2 by day 2 of storage (which was reduced to 4.6 log CFU/cm2 by day 14), while the total microbial populations showed more extensive attachment (6.1 to 6.6 log CFU/cm2). The sanitizers were more effective in reducing populations of cells in suspension than in reducing populations of attached cells. Overall, there were no differences between previously acid-adapted and nonadapted L monocytogenes with regard to sensitivity to sanitizers. The total microbial biofilms were the most sensitive to all of the sanitizers on day 2, but their resistance increased during storage, and they were at their most resistant on day 14. Listeria monocytogenes displayed stronger resistance to the effects of the sanitizers on day 7 than on day 2 but had become sensitized to all sanitizers by day 14. SH at the adjusted pH (6.80) (ASH) was generally more effective in reducing bacterial populations than was SH at the unadjusted pH. PAA generally killed attached cells faster at 30 to 300 s of exposure than did the other sanitizers, except for ASH on day 2. PAA was more effective in killing attached cells than in killing cells treated in suspension, in contrast to the other sanitizers.     

Combination of high-intensity pulsed electric fields with natural antimicrobials to inactivate pathogenic microorganisms and extend the shelf-life of melon and watermelon juices

The effect of high-intensity pulsed electric field (HIPEF) combined with citric acid (0.5-2.0%, w/v) or cinnamon bark oil (0.05-0.30%, w/v) against populations of Escherichia coli O157:H7, Salmonella Enteritidis and Listeria monocytogenes in melon and watermelon juices were evaluated. Microbiological shelf-life and sensory attributes were also determined. Populations of E. coli O157:H7, S. Enteritidis and L. monocytogenes were reduced by more than 5.0log(10)CFU/ml in HIPEF-processed melon (35kV/cm for 1709 micros at 193Hz and 4 micros pulse duration) and watermelon (35kV/cm for 1682 micros at 193Hz and 4 micros pulse duration) juices containing 2.0% and 1.5% of citric acid, respectively, or 0.2% of cinnamon bark oil. In addition, these treatments were also able to inactivate mesophilic, psychrophilic and, molds and yeasts populations, leading to a shelf-life of more than 91 days in both juices stored at 5 degrees C. Hence, the microbiological quality and safety of these fruit juices by combining HIPEF and citric acid or cinnamon bark oil were ensured. However, the taste and odor in those HIPEF-treated melon and watermelon juices containing antimicrobials were significantly affected. Therefore, further studies are needed to decrease the impact on the sensory attributes by using antimicrobials.     

INHIBITION OF LISTERIA MONOCYTOGENES BY CARNOBACTERIUM PISCICOLA IN VACUUM-PACKAGED COOKED CHICKEN AT REFRIGERATION TEMPERATURES

Listeria monocytogenes inhibition by bacteriocinogenic Carnobacterium piscicola DX or nonbacteriocinogenic C. piscicola 2818 was examined in vacuum-packed minimally processed chicken cubes with gravy at 4, 8 and 15C. C. piscicola DX and C. piscicola 2818 at 10⁴ CFU/g were coinoculated individually with a pool of three Listeria monocytogenes strains at 10² CFU/g. At 4 and 8C , C. piscicola DX inhibited growth of L. monocytogenes by 3 logs, significantly (P < 0.05) more than did C. piscicola 2818. At 15C both C. piscicola strains inhibited L. monocytogenes by one log cycle. The pH of all inoculated systems decreased from an initial pH of 6.14 to final values ranging from 6.06 to 5.65 depending on the inocula used. Bacteriocin was detected in the systems coinoculated with C. piscicola DX. These studies demonstrate that lower temperatures and bacteriocin production enhanced L. monocytogenes inhibition by C. piscicola     

Reduction of Listeria monocytogenes and Escherichia coli O157:H7 numbers on vacuum-packaged fresh beef treated with nisin or nisin combined with EDTA.

Nisin or nisin combined with EDTA was used to treat fresh beef. Beef cubes (2.5 by 2.5 by 2.5 cm) that were inoculated with approximately 7 log CFU/ml of Listeria monocytogenes Scott A or Escherichia coli O157:H7 505 B were dipped in the following solutions: (i) H2O, (ii) HCl, (iii) nisin, (iv) EDTA, or (v) nisin combined with EDTA, respectively, for 10 min each, with an exception of one set of control beef samples without treatment. Beef samples were then drip-dried for 15 min, vacuum packaged, and stored at 4 degrees C for up to 30 days. The pH on beef after different treatments was not a key factor in preventing bacterial growth. Treatment with nisin or with nisin combined with EDTA reduced the population of L. monocytogenes by 2.01 and 0.99 log CFU/cm2 as compared to the control, respectively, under the conditions of vacuum package and storage at 4 degrees C for up to 30 days. However, the effect of nisin and nisin combined with EDTA against E. coli O157:H7 505 B was marginal at 1.02 log CFU/cm2 and 0.8 log CFU/cm2 reductions, respectively.     

Growth of Listeria monocytogenes in egg salad and pasta salad formulated with mayonnaise of various pH and stored at refrigerated and abuse temperatures

This study investigated and modeled the behavior of Listeria monocytogenes in egg salad and pasta salad as affected by mayonnaise pH (3.8, 4.2, 4.6, and 5.0) and storage temperature (4, 8, and 12 degrees C). At each storage temperature, L. monocytogenes was able to grow in both salads regardless of the mayonnaise pH. The lag-phase durations (LPD) of L. monocytogenes in egg salad ranged from 33 to 85, 15 to 50, and 0 to 19 h, and the growth rates (GR) ranged from 0.0187 to 0.0318, 0.0387 to 0.0512, and 0.0694 to 0.1003 log(10)cfu/h at 4, 8, and 12 degrees C, respectively. The LPD of L. monocytogenes in pasta salad ranged from 210 to 430, 49 to 131, and 21 to 103 h, and GR ranged from 0.0118 to 0.0350, 0.0153 to 0.0418, and 0.0453 to 0.0718 log(10)cfu/h at 4, 8, and 12 degrees C, respectively. The growth of L. monocytogenes was more rapid in egg salad than in pasta salad, indicating that a better growth environment for L. monocytogenes existed in egg salad. In both salads, the LPD decreased and the GR increased as the storage temperature increased. Mathematical models and response surface plots describing the LPD and GR of L. monocytogenes in both salads as affected by the mayonnaise pH and storage temperature were developed. The models confirmed that the growth of L. monocytogenes in egg salad and pasta salad was primarily promoted by higher storage temperatures and, secondarily, by higher mayonnaise pH. The conditions under which the models may be applied to estimate the growth of L. monocytogenes in both salads were identified.     

Application of reuterin produced by Lactobacillus reuteri 12002 for meat decontamination and preservation

Lactobacillus reuteri strain 12002 was used for reuterin production in the two-step fermentation process. A batch culture fermentation was used to produce a maximum biomass of L. reuteri. Then cells were harvested, resuspended in a glycerol-water solution, and anaerobically incubated to produce reuterin. The lyophilized supernatants (approximately 4000 activity units (AU) of reuterin per ml) were diluted in distilled water for decontamination and preservation trials. The MIC values of reuterin for Escherichia coli O157:H7 and Listeria monocytogenes were 4 and 8 AU/ml, respectively. In meat decontamination experiments, the surface of cooked pork was inoculated with either L. monocytogenes or E. coli O157:H7 at a level of approximately log10 5 CFU/cm2, incubated for 30 min at 7 degrees C, and decontaminated by exposure to reuterin (500 AU/ml). The bactericidal effect of reuterin was analyzed 15 s and 24 h after exposure at 7 degrees C. After 15 s of exposure to reuterin, viable numbers decreased by 0.45 and 0.3 log10 CFU/cm2 for E. coli O157:H7 and L. monocytogenes, respectively. After 24 h the numbers decreased by 2.7 log10 CFU/cm2 for E. coli O157:H7 and by 0.63 log10 CFU/cm2 for L. monocytogenes. In the same experiment, the combined effect of reuterin and lactic acid was also investigated. Adding lactic acid (5%, vol/vol) to reuterin significantly enhanced (P < or = 0.05) the efficacy of reuterin. No additional effect (P < or = 0.05) was found when ethanol (40%) was added to the mixture of reuterin and lactic acid. To evaluate the preservative effect of reuterin during meat storage, reuterin was added to raw ground pork contaminated with E. coli O157:H7 or L. monocytogenes. Reuterin at a concentration of 100 AU/g resulted in a 5.0-log10 reduction of the viability of E. coli O157:H7 after 1 day of storage at 7 degrees C. Reuterin at a concentration of 250 AU/g reduced the number of the viable cells of L. monocytogenes by log10 3.0 cycles after 1 week of storage at 7 degrees C.     

Effect of acid tolerance response (ATR) on attachment of Listeria monocytogenes Scott A to stainless steel under extended exposure to acid or/and salt stress and resistance of sessile cells to subsequent strong acid challenge

The aim of this study was to investigate the potential effect of adaptive stationary phase acid tolerance response (ATR) of Listeria monocytogenes Scott A cells on their attachment to stainless steel (SS) under low pH or/and high salt conditions and on the subsequent resistance of sessile cells to strong acid challenge. Nonadapted or acid-adapted stationary-phase L. monocytogenes cells were used to inoculate (ca. 10? CFU/ml) Brain Heart (BH) broth (pH 7.4, 0.5% w/v NaCl) in test tubes containing vertically placed SS coupons (used as abiotic substrates for bacterial attachment). Incubation was carried out at 16 °C for up to 15 days, without any nutrient refreshment. L. monocytogenes cells, prepared as described above, were also exposed to low pH (4.5; adjusted with HCl) or/and high salt (5.5% w/v NaCl) stresses, during attachment. On the 5th, 10th and 15th day of incubation, cells attached to SS coupons were detached (through bead vortexing) and enumerated (by agar plating). Results revealed that ATR significantly (p<0.05) affected bacterial attachment, when the latter took place under moderate acidic conditions (pH 4.5, 0.5 or 5.5% w/v NaCl), with the acid-adapted cells adhering slightly more than the nonadapted ones. Regardless of acidity/salinity conditions during attachment, ATR also enhanced the resistance of sessile cells to subsequent lethal acid challenge (exposure to pH 2 for 6 min; pH adjusted with either hydrochloric or lactic acid). The trend observed with viable count data agreed well with conductance measurements, used to indirectly quantify remaining attached bacteria (following the strong acid challenge) via their metabolic activity. To sum, this study demonstrates that acid adaptation of L. monocytogenes cells during their planktonic growth enhances their subsequent attachment to SS under extended exposure (at 16 °C for up to 15 days) to mild acidic conditions (pH 4.5), while it also improves the resistance of sessile cells to extreme acid treatment (pH 2). Therefore, the ATR of bacterial cells should be carefully considered when applying acidic decontamination strategies to eradicate L. monocytogenes attached to food processing equipment.     

Impact of the population of spoilage microflora on the growth of Listeria monocytogenes on frankfurters

Approximately 100 CFU/cm2 of a five-strain mixture of Listeria monocytogenes was coinoculated onto frankfurters with three different concentrations (10(2), 10(4), and 106 CFU/cm2) of an undefined spoilage microflora derived from commercial frankfurters. The frankfurters were vacuum packaged and stored at 10 degrees C for up to 48 days. The populations of L. monocytogenes, aerobic mesophilic bacteria, lactic acid bacteria, and Enterobacteriaceae were determined at various time intervals during storage. After 14 days, the population of L. monocytogenes was highest when grown with a spoilage microflora population of 10(2) CFU/cm2, and this trend continued until 48 days. Throughout the entire storage period, the populations of L. monocytogenes at any concentration of inoculated spoilage microflora rarely differed by more than 0.5 log CFU/cm2, and the maximum observed difference as 1.1 log CFU/cm2 at 40 days. The growth rate of L. monocytogenes was approximately the same at all concentrations of the inoculated spoilage microflora. These results suggest that the concentration of spoilage microflora present on the original processed meat may have a slight impact on the growth of L. monocytogenes in the package.     

脉冲电场强化天冬酰胺- 果糖体系的研究

应用脉冲电场强化天冬酰胺- 果糖模式美拉德反应,对反应后天冬酰胺- 果糖模式美拉德反应体系的pH 值、紫外特征吸收(A294nm)、褐变程度(A420nm)、反应过程中天冬酰胺和果糖含量以及抗氧化活性进行分析,同时采用高效液相色谱(HPLC)检测产物中丙烯酰胺和5- 羟甲基糠醛(HMF)的含量。结果表明：脉冲电场能显著促进天冬酰胺-果糖模式美拉德反应,体系褐变程度、抗氧化活性与反应时间及电场强度均呈正相关,反应产物中未发现丙烯酰胺和HMF。     

Influence of poultry carcass skin sample site on the effectiveness of trisodium phosphate against Listeria monocytogenes

The aim of this study was to determine the influence of skin sample site on the efficacy of trisodium phosphate (TSP) solutions in reducing Listeria monocytogenes populations on chicken carcasses during refrigerated storage. Chicken skin samples from the legs, the breasts, and the dorsal area inoculated with L. monocytogenes (10(8) CFU/ml) were dipped for 15 min in sterile tap water (control) or in 8, 10, or 12% TSP. L. monocytogenes counts and surface pH values were determined after 0, 1, 3, and 5 days of storage at 2 degrees C. For all sampling times and TSP concentrations, the reductions in L. monocytogenes numbers in breast skin were significantly larger (P < 0.05) than those in leg skin or dorsal skin. No significant differences were found in pH values as an effect of skin site. Our results suggest that skin sampling site is an important factor that needs to be considered when decontamination protocols are developed for poultry carcasses with the TSP treatment.     

Effect of acid adaptation on growth during storage at 10 degrees C and resistance to simulated gastric fluid of Listeria monocytogenes inoculated onto bologna formulated with or without antimicrobials

The fate of acid-adapted and nonadapted Listeria monocytogenes inoculated onto bologna slices (formulated with or without antimicrobials) was examined during storage and after exposure to in vitro gastric challenge. Bologna slices formulated with no antimicrobials (control), 3% sodium lactate (SL), or 1.8% SL plus 0.25% sodium diacetate (SD) were inoculated (2 log CFU/cm2) with a 10-strain composite of acid-adapted or nonadapted L. monocytogenes strains. Growth or survival of the two inocula on bologna was evaluated during vacuum-packaged storage (10 degrees C) for up to 36 days. Survival of previously acid-adapted or nonadapted L. monocytogenes on stored bologna exposed to simulated gastric fluid (adjusted to pH 1.0 with HCl) for 20, 40, and 60 min also was determined. As expected, inclusion of antimicrobials in the product formulation inhibited growth of L. monocytogenes during storage of vacuum-packaged bologna compared with growth on control samples. Acid adaptation of L. monocytogenes prior to product inoculation did not affect subsequent survival or growth on bologna or resistance to simulated gastric fluid (P > 0.05). Survival of L. monocytogenes exposed to simulated gastric fluid during storage increased with product age, growth phase of the cells, and possibly age of the cells, particularly for control samples (no antimicrobials), in which the pathogen grew uninhibited to approximately 6 log CFU/cm2 by day 8 of storage. Inhibition of L. monocytogenes growth on product formulated with antimicrobials was associated with only sporadic and small numbers of survivors following exposure of these samples to simulated gastric fluid, especially in samples stored longer. However, cell numbers in these treatment groups before the gastric challenge did not exceed 3.8 log CFU/cm2. Inhibition of growth on product with antimicrobials precluded detection of survivors resistant to the effects of simulated gastric fluid.