A predictive model to determine the effects of temperature,sodium pyrophosphate,and sodium chloride on thermal inactivation of starved Listeria monocytogenes in pork slurry

The effects and interactions of 27 combinations of heating temperature (57.5 to 62.5 degrees C), sodium pyrophosphate (SPP) level (0 to 0.5%, wt/vol), and salt (NaCl) level (0 to 6%, wt/vol) on the thermal inactivation of starved Listeria monocytogenes ATCC 19116 in pork slurry were investigated. A split-split plot experimental design was used to compare all 27 combinations. L. monocytogenes survivors were enumerated on tryptic soy agar supplemented with 0.6% yeast extract. The natural logarithm (loge) of the means of decimal reduction times (D-values) were modeled as a function of temperature, SPP level, and NaCl level. Increasing concentrations of SPP or NaCl protected starved L. monocytogenes from the destructive effect of heat. For example, D-values for the pathogen at 57.5 degrees C in pork slurry with 0, 3, and 6% NaCl were 2.79, 7.75, and 14.59 min, respectively. All three variables interacted to affect the thermal inactivation of L. monocytogenes. A mathematical model describing the combined effect of temperature, SPP level, and NaCl level on the thermal inactivation of starved L. monocytogenes was developed. There was strong correlation (R2 = 0.97) between loge D-values predicted by the model and those observed experimentally. The model can predict D-values for any combination of variables that falls within the range of those tested. This predictive model can be used to assist food processors in designing thermal processes that include an adequate margin of safety for the control of L. monocytogenes in processed meats.     

Predictive model for the combined effect of temperature,sodium lactate,and sodium diacetate on the heat resistance of Listeria monocytogenes in beef

The effects of heating temperature (60 to 73.9 degrees C), sodium lactate (NaL; 0.0 to 4.8% [wt/wt]), and/or sodium diacetate (SDA; 0.0 to 0.25% [wt/wt]) and of the interactions of these factors on the heat resistance of a five-strain mixture of Listeria monocytogenes in 75% lean ground beef were examined. Thermal death times for L. monocytogenes in filtered stomacher bags in a circulating water bath were determined. The recovery medium was tryptic soy agar supplemented with 0.6% yeast extract and 1% sodium pyruvate. Decimal reduction times (D-values) were calculated by fitting a survival model to the data with a curve-fitting program. The D-values were analyzed by second-order response surface regression for temperature, NaL level, and SDA level. The D-values observed for beef with no NaL or SDA at 60, 65, 71.1, and 73.9 degrees C were 4.67, 0.72, 0.17, and 0.04 min, respectively. The addition of 4.8% NaL to beef increased heat resistance at all temperatures, with D-values ranging from 14.3 min at 60 degrees C to 0.13 min at 73.9 degrees C. Sodium diacetate interacted with NaL, thereby reducing the protective effect of NaL and rendering L. monocytogenes in beef less resistant to heat. A mathematical model describing the combined effect of temperature, NaL level, and SDA level on the thermal inactivation of L. monocytogenes was developed. This model can predict D-values for any combination of temperature, NaL level, and SDA level that is within the range of those tested. This predictive model will have substantial practical importance to processors of cooked meat, allowing them to vary their thermal treatments of ready-to-eat meat products in a safe manner.     

PREDICTIVE MODEL FOR THE COMBINED EFFECT OF TEMPERATURE,pH, SODIUM CHLORIDE,AND SODIUM PYROPHOSPHATE ON THE HEAT RESISTANCE OF ESCHERICHIA COLI O157:H71

The effects and interactions of heating temperature (5–62.5C), pH (4 – 8), NaCl (0 – 6%, w/v), and sodium pyrophosphate (0 – 0.3%, w/v) on the heat resistance of a four strain mixture of Escherichia coli O157:H7 in beef gravy were examined. Thermal death times were determined using a submerged coil heating apparatus. The recovery medium was plate count agar supplemented with 1% sodium pyruvate. Decimal reduction times (D-values) were calculated by fitting a survival model to the data with a curve fitting program. The D-values were analyzed by second order response surface regression for temperature, pH, NaCl and sodium pyrophosphate levels. The four variables interacted to affect the inactivation of the pathogen. Thermal resistance of E. coli O157:H7 can be lowered by combining these intrinsic factors. A mathematical model describing the combined effect of temperature, pH, NaCl and sodium pyrophosphate levels on the thermal inactivation of E. coli O157:H7 was developed. The model can predict D-values for any combinations of temperature, pH, NaCl and sodium pyrophosphate that are within the range of those tested.     

Modeling the effect of storage atmosphere on growth-no growth interface of Listeria monocytogenes as a function of temperature, sodium lactate, sodium diacetate, and NaCl

The effect of aerobic and anaerobic conditions on growth initiation by a 10-strain composite of Listeria monocytogenes (10(4) CFU/ml) was evaluated in tryptic soy broth with 0.6% yeast extract (TSBYE) as a function of 220 combinations of pH (3.82 to 7.42), sodium lactate (SL) (0 to 10%, vol/vol), and sodium diacetate (SD) (0 to 0.5%, wt/vol) at 10 or 30 degrees C (a slightly abusive and the optimal growth temperature, both above the growth limiting range of 0 to 3 degrees C for L. monocytogenes) in 96-well microplates. In addition, four probability-of-growth models were developed to quantify the effect of 346 aerobic and 346 anaerobic combinations of temperature (4 to 30 degrees C), SL (0 to 6%, vol/vol), and SD (0 to 0.5%, wt/vol) in the presence of NaCl (0.5 or 2.5%, wt/vol) on the growth-no growth responses of the same L. monocytogenes strain composite, with a microplate reader. Growth responses were evaluated turbidimetrically (620 nm) every 5 days for a total of 40 days. Data were modeled with logistic regression to determine the growth-no growth interfaces. The minimum pH values at which growth of L. monocytogenes occurred were higher under anaerobic than under aerobic conditions, and this difference was more evident at 10 degrees C or at higher SL and SD concentrations. The MIC of SD decreased with increasing SL levels. Anaerobic storage reduced the levels of SL-SD, allowing the growth of L. monocytogenes compared with aerobic storage, especially at low temperatures. In the presence of 2.5% NaCl, the MICs for SD were lower than those obtained with 0.5% NaCl, especially at 4 and 10 degrees C, or in the presence of 5 to 6% SL. The developed models for anaerobic incubation showed good performance (80% successful predictions; i.e., in 40 of 50 comparisons) with independent data from studies on survival-growth of L. monocytogenes on meat products. The study provides quantitative data on the antimicrobial activity of SL (0 to 10%) and SD (0 to 0.5%), temperature (4 to 30 degrees C), and pH (3.82 to 7.42) and on the probability of growth of L. monocytogenes under anaerobic or aerobic conditions in the presence of 0.5 or 2.5% NaCl, and hence, addresses important needs for risk assessment activities.     

Effect of sodium lactate on thermal inactivation of Listeria monocytogenes and Salmonella in ground chicken thigh and leg meat

The effect of sodium lactate on thermal inactivation D- and z-values of Listeria monocytogenes and Salmonella was determined for chicken thigh and leg meat. At 55 to 70 degrees C, the D-value of L. monocytogenes in ground chicken thigh and leg meat with the addition of 4.8% sodium lactate (4.8 g sodium lactate per 100 g of meat) was 53 to 75% higher than that in the meat without sodium lactate. No significant difference was found for the D-values of Salmonella at 55 to 70 degrees C between the meat with and that without sodium lactate (4.8%. wt/wt). The z-values of both L. monocytogenes and Salmonella were not affected by sodium lactate (4.8%). The results from this study are useful for predicting thermal process lethality of L. montocytogenes and Salmonella in formulated chicken thigh and leg meat products.     

Acid and NaCl limits to growth of Listeria monocytogenes and influence of sequence of inimical acid and NaCl levels on inactivation kinetics

Variability in growth limits of Listeria monocytogenes in response to low pH (adjusted with HCl) or high salinity (NaCl) was evaluated for 127 strains in brain heart infusion broth at 25 degrees C. Over 95% of strains habituated at pH 5.0 grew subsequently at pH 4.2, while 25% were able to grow at pH 4.1. More than 85% of strains preadapted to growth at 8.5% NaCl (wt/vol) subsequently grew in the presence of 11.3% NaCl, while 25% were able to grow at 13% NaCl, and 4.7% grew in the presence of 13.9% NaCl. The results extend the generally accepted growth limits for L. monocytogenes in response to these hurdles. Two strains, one of which was relatively tolerant of both hurdles, and another that was less tolerant of both hurdles, were subjected to different sequences of lethal acid (pH 3.5) and NaCl (14%, wt/vol) stresses to determine whether survival was affected by growth limits, or by sequence of application of treatment. There was no significant difference in the inactivation kinetics of the two strains, but inactivation rates were affected by different treatments. For both strains, the inactivation rates, from fastest to slowest, resulted from: (i) lethal pH and then by lethal water activity, or lethal water activity and then by lethal pH; (ii) lethal pH and water activity applied simultaneously; (iii) lethal pH; and (iv) lethal water activity. The results demonstrated that the sequence of lethal stress application strongly influences L. monocytogenes inactivation, and that L. monocytogenes growth limits are not good predictors of survival in inimical environments.     

INFLUENCE OF THE INTRINSIC PROPERTIES OF FOOD ON THERMAL INACTTVATION OF SPORES OF NONPROTEOLYTIC CLOSTRIDIUM BOTULINUM: DEVELOPMENT OF A PREDICTIVE MODEL1

The effects and interactions of heating temperature (70–90C), pH (5–6.5), sodium chloride (0–3%), and sodium pyrophosphate (0–0.3%) on the heat resistance of a six strain mixture of spores of nonproteolytic Clostridium botulinum type B and type E in turkey were examined. Thermal death times were determined in submerged vials heated using a water bath. Heated spores were recovered on Reinforced Clostridial Medium (RCM) supplemented with lysozyme (10 μg/ml). Decimal reduction times (D-values) were calculated by fitting a survival model to the data with a curve fitting program. The D-values were analyzed by second order response surface regression for temperature, pH, salt (sodium chloride) and sodium pyrophosphate levels. The four variables interacted to effect the inactivation of spores. Confidence intervals (95%) predicted heat resistance of spores in turkey. The data suggest that the effect of reduced pH in increasing the inactivation was more pronounced at high temperatures and may provide an adequate degree of protection from nonproteolytic C. botulinum spores in minimally processed foods, particularly if employed in conjunction with combinations of salt and sodium pyrophosphate.     

A predictive model to describe the effects of temperature, sodium lactate, and sodium diacetate on the inactivation of a serotype 4b strain of Listeria monocytogenes in a frankfurter slurry

A modified Gompertz equation was used to model the effects of temperature (55, 60, and 65 degrees C), sodium lactate (0, 2.4, and 4.8%), and sodium diacetate (0, 0.125, and 0.25%) on inactivation of Listeria monocytogenes strain MFS 102 (serotype 4b) in frankfurter slurry. The effects of these factors were determined on the shouldering region (parameter A), maximum death rate (parameter B), and tailing region (parameter C) of microbial inactivation curves. Increased temperature or sodium diacetate concentrations increased the death rate, whereas increased sodium lactate concentrations decreased heat resistance. Complex two-way interactive effects were also observed. As both temperature and sodium lactate increased, the death rate decreased; however, as temperature and sodium diacetate increased, the death rate increased. The effect of the interaction between sodium lactate and sodium diacetate on the maximum death rate varied with temperature. Increases in both acidulants at temperatures above 56.7 degrees C decreased the death rate, whereas at temperatures below 56.7 degrees C, increases in both acidulants increased the death rate. To test for significant differences between treatments, D-values were calculated and compared. This comparison revealed that, in general, sodium lactate increased heat resistance and sodium diacetate decreased heat resistance of L. monocytogenes. This information is important for reducing and minimizing contamination during postprocessing thermal treatments.     

Thermal resistance of Salmonella enterica serotypes, Listeria monocytogenes, and Staphylococcus aureus in high solids liquid egg mixes

Decimal reduction times (D-values) were determined for Salmonella enterica serotypes, Listeria monocytogenes, and Staphylococcus aureus in two high solids egg mixes designated A and B (water activity [a(w)] = 0.76 and 0.82; solids = 53.12 and 52.63%; pH = 5.09 and 5.29; viscosity = 183 and 119 centipoise/s, respectively) using a low-volume (0.06 ml) sealed glass capillary tube procedure. For Salmonella, D-values ranged from 0.035 (70 degrees C) to 0.193 min (64 degrees C) in product A and from 0.048 to 0.193 min in product B. For Listeria, D-values ranged from 0.133 (70 degrees C) to 0.440 min (64 degrees C) in product A and from 0.074 to 0.364 min in product B. For Staphylococcus, D-values ranged from 0.332 (70 degrees C) to 1.304 min (64 degrees C) in product A and from 0.428 to 1.768 min in product B. For Listeria, the D-values of all heating temperatures were significantly higher (P < 0.01) in product A than in product B. The similar trend was also observed for Salmonella and Staphylococcus but only at 66 degrees C for Salmonella and 64 degrees C for Staphylococcus. Greater temperature dependence was observed for Salmonella inactivation in the low a(w) and low pH product (A), while the product (B) with the higher a(w) and pH had greater temperature dependence for Listeria. Compared across both egg mixes and all heating temperatures, the Staphylococcus strains were from 6.2 to 11.7 times more heat resistant than S. enterica serotypes and from 2.2 to 7.5 times more heat resistant than L. monocytogenes.     

Modelling the effect of pH,sodium chloride and sodium pyrophosphate on the thermal resistance of Escherichia coli O157:H7 in ground beef

The objective of this study was to assess the combined effects of temperature, pH, sodium chloride (NaCl), and sodium pyrophosphate (SPP) on the heat resistance of Escherichia coli O157:H7 in minced beef meat. A fractional factorial design consisted of four internal temperatures (55.0, 57.5, 60.0 and 62.5 °C), five concentrations of NaCl (0.0, 1.5, 3.0, 4.5 and 6.0 wt/wt.%) and SPP (0.0, 0.1, 0.15, 0.2 and 0.3 wt/wt.%), and five levels of pH (4.0, 5.0, 6.0, 7.0 and 8.0). The 38 variable combinations were replicated twice to provide a total of 76 survivor curves, which were modelled by a modified three-parameter Weibull function as primary model. The polynomial secondary models, developed to estimate the time to achieve a 3-log and a 5-log reduction, enabled the estimation of critical pH, NaCl and SPP concentrations, which are values at which the thermo-tolerance of E. coli O157:H7 reaches it maximum. The addition up to a certain critical concentration of NaCl (~ 2.7–4.7%) or SPP (~ 0.16%) acts independently to increase the heat resistance of E. coli O157:H7. Beyond such critical concentrations, the thermo-resistance of E. coli O157:H7 will progressively diminish. A similar pattern was found for pH with a critical value between 6.0 and 6.7, depending upon temperature and NaCl concentration. A mixed-effects omnibus regression model further revealed that the acidity of the matrix and NaCl concentration had a greater impact on the inactivation kinetics of E. coli O157:H7 in minced beef than SPP, and both are responsible for the concavity/convexity of the curves. When pH, SPP or NaCl concentration is far above or below from its critical value, the temperatures needed to reduce E. coli O157:H7 up to a certain log level are much lower than those required when any other environmental condition is at its critical value. Meat processors can use the model to design lethality treatments in order to achieve specific log reductions of E. coli O157:H7 in ready-to-eat beef products.     

Effects and interactions of sodium lactate, sodium diacetate, and pediocin on the thermal inactivation of starved Listeria monocytogenes on bologna

The effects and interactions of temperature (56.3-60 °C), sodium lactate (SL; 0-4.8%), sodium diacetate (SDA; 0-2.5%), and pediocin (0-10,000 AU) on starved Listeria monocytogenes (10⁷ CFU/g) on bologna were investigated. Bologna slices containing SL and SDA in the formulation were dipped in pediocin, surface inoculated, and treated at various temperatures using combinations of parameters determined by central composite design. D-values were calculated. The observed D-values ranged from 2.8 min at 60 °C to 24.61 min at 56.3 °C. Injury ranged from 9.1 to 76% under various conditions. The observed D-values were analyzed using second order response surface regression for temperature, SL, SDA, and pediocin, and a predictive model was developed. Predicted D-values were calculated and ranged from 3.7 to 19 min for various combinations of parameters. Temperature alone reduced the predicted D-values from 33.96 min at 56.3 °C to 11.51 min at 60 °C. Addition of SL showed a protective effect. Other combination treatments either reduced or increased D-values depending on temperature. The combination of SL and SDA was effective at lower temperatures, however, higher levels of SDA at higher temperatures made the organism more heat resistant. Pediocin (up to 5000 AU) with increasing temperature and SDA reduced D-values. Depending on temperature and concentration, the interactions between various additives can affect thermal inactivation of L. monocytogenes on bologna. Starvation rendered L. monocytogenes more susceptible to heat and additives.     

Sodium lactate,sodium diacetate and pediocin: Effects and interactions on the thermal inactivation of Listeria monocytogenes on bologna

The effects and interactions of temperature (56.3–60 °C), sodium lactate (SL; 0–4.8%), sodium diacetate (SD; 0–0.25%) and pediocin (0–10,000 AU) on Listeria monocytogenes on bologna were studied and a predictive inactivation model was developed. Bologna was manufactured with different SL/SD concentrations in the formulation, dipped in pediocin solution and treated at different temperatures using combinations of parameters determined by central composite design. D-values were calculated and analyzed using second order response regression. Predicted D-values were also calculated. The observed D-values for L. monocytogenes on bologna ranged from 2.10 to 35.59 min. Temperature alone decreased predicted D-values from 99.02 min at 56.3 °C to 44.71 min at 60.0 °C. Adding SL decreased D-values (85.43–22.71 min) further; however, heat and SD combined was the most effective for reducing L. monocytogenes on bologna. An SD level of 0.25% at 58.2 °C had the overall lowest predicted D-value (15.95 min). Combination treatments increased or decreased D-values, depending on the temperature. Pediocin (2500 and 5000 AU) and heat decreased D-values, but exhibited a protective effect at higher concentrations (≥7500 AU). The results showed that interactions between additives in formulations can vary at different temperatures/concentrations, thereby affecting thermal inactivation of foodborne pathogens in meat products. Hence, food processors should modify food formulations carefully, and verify with adequate testing so that product safety is not compromised.     

Heat and acid tolerance of Listeria monocytogenes after exposure to single and multiple sublethal stresses

The majority of published studies on the adaptive heat or acid tolerance response of Listeria monocytogenes have been performed with a single strain exposed to a single adaptation treatment; however, in food ecosystems, microorganisms commonly exist as multi-species communities and encounter multiple stresses, which may result in "stress hardening". Therefore, the present study evaluated the adaptive responses to heat (52, 57 and 63 degrees C) or lactic acid (pH 3.5) of a 10-strain composite of L. monocytogenes meat and human isolates at stationary phase, following exposure to combinations of osmotic (10% NaCl), acidic (pH 5.0 with HCl) and thermal (T; 46 degrees C) stresses, sequentially or simultaneously within 1.5h, in tryptic soy broth with 0.6% yeast extract (TSBYE). All treatments induced adaptive responses on L. monocytogenes at 57 degrees C, while no such cross-protection was observed at 52 and 63 degrees C. Survivor curves at 57 degrees C appeared convex with profound shoulders determined by a Weibull model. The highest thermotolerance was observed after combined exposure to acid and heat shock (pH-T), followed by exposure to osmotic shock, and by the combination of osmotic with heat shock (NaCl-T). Regarding acid tolerance, prior exposure to low pH, pH-T, or a combination of NaCl, pH and T resulted in a marked increase of resistance to pH 3.5, showing concave inactivation curves with tails at higher levels of survivors (log(10)CFU ml(-1)) than the control cultures. The sequence of exposure to sublethal stresses did not affect the thermotolerance of L. monocytogenes, whereas simultaneous exposure to most multiple stresses (e.g., NaCl-pH-T, NaCl-T and NaCl-pH) resulted in higher survivors of L. monocytogenes at pH 3.5 than exposure to the same stresses sequentially. The results indicate that combinations and sequences of sublethal hurdles may affect L. monocytogenes acid and heat tolerance, especially in acidic environments with mild heating or in low moisture environments.     

Radiation-heat synergism for inactivation of Alicyclobacillus acidoterrestris spores in citrus juice

The objective of this study was to investigate the influence of electron-beam and gamma-ray irradiation and temperature (85 to 95 degrees C) on Alicyclobacillus acidoterrestris GD3B strain (NCIMB 13137) spores by calculating and comparing the decimal reduction dose or time (D-values). The survival rate of A. acidoterrestris spores decreased exponentially with irradiation doses of an electron beam or gamma ray. D-values determined for electron-beam and gamma-ray irradiated spores on filter paper ranged from 1.02 to 1.10 kGy. On the other hand, the thermal sterilization effect showed a single exponential decrease within 1.5-log decreases in cell numbers (D85 degrees C = 70.5 min, D90 degrees C = 16.1 min, and D95 degrees C = 5.19 min and z-value [change in temperature required to change the D-value] was 8.83 degrees C), and prolonged heating produced an increase of 10 to 13 times that of the thermal resistance. However, within all time ranges studied (5 to 360 min), a linear decrease in the D-value was observed with an increase in the temperature. A combination of two different methods, irradiation before heating, was appropriate for reducing the duration of the heat treatment required to achieve the inactivation of conidia. Moreover, a necessary radiation dosage for complete inactivation of A. acidoterrestris spores that contaminated dextrin was examined. Dextrin is often used in the juice industry as an augmentor, and it is known to be sometimes contaminated by these spores. The D-values of the spores in dextrin for electron-beam and gamma-ray irradiations were 1.72 and 1.79 kGy, respectively. The doses required for elimination of the spores could be lowered by using irradiation in combination with heat sterilization. When dextrin powder contaminated with 10(4) CFU/g of A. acidoterrestris was preirradiated at 1.0 kGy of electron beam, the citrus juice containing dextrin at a concentration of 10% (wt/vol) was completely sterilized by heating for 20 min at 95 degrees C.     

Short communication: Salt tolerance of Lactococcus lactis R-604 as influenced by mild stresses from ethanol,heat, hydrogen peroxide,and UV light

Lactococcus lactis is a culture widely used in salt-containing dairy products. Salt hinders bacterial growth, but exposure to environmental stress may protect cells against subsequent stress, including salt. The objective of this study was to evaluate the salt tolerance of L. lactis R-604 after exposure to various stresses. The culture was subjected to 10% (vol/vol) ethanol for 30 min, mild heat at 52°C for 30 min, 15 mM hydrogen peroxide for 30 min, or UV light (254 nm) for 5 min and compared with a control. Starting with 5 log cfu/mL for all treatments, growth was determined in M17 broth with 5 NaCl concentrations (0, 1, 3, 5, and 7% wt/vol). Plating was conducted daily for 5 d. Salt tolerance was enhanced with mild heat exposure before growth in M17 broth with 5% (wt/vol) NaCl on d 3, 4, and 5, and with exposure to hydrogen peroxide and ethanol stresses before growth in M17 broth with 5% (wt/vol) NaCl on d 4 and 5. Exposure of this culture to mild heat, hydrogen peroxide, or ethanol before growth in M17 broth containing 5% (wt/vol) salt can enhance its survival, which could be beneficial when using it in salt-containing dairy products.     

Studies on the pathogenesis and survival of different culture forms of Listeria monocytogenes to pulsed UV-light irradiation after exposure to mild-food processing stresses

The effects of mild conventional food-processing conditions on Listeria monocytogenes survival to pulsed UV (PUV) irradiation and virulence-associated characteristics were investigated. Specifically, this study describes the inability of 10 strains representative of 3 different culture forms or morphotypes of L. monocytogenes to adapt to normally lethal levels of PUV-irradiation after exposure to sub-lethal concentrations of salt (7.5% (w/v) NaCl for 1 h), acid (pH 5.5 for 1 h), heating (48 °C for 1 h) or PUV (UV dose 0.08 μJ/cm(2)). Findings showed that the order of increasing sensitivity of L. monocytogenes of non-adapted and stressed morphotypes to low pH (pH 3.5 for 5 h, adjusted with lactic), high salt (17.5% w/v NaCl for 5 h), heating (60 °C for 1 h) and PUV-irradiation (100 pulses at 7.2 J and 12.8 J, equivalent to UV doses of 2.7 and 8.4 μJ/cm(2) respectively) was typical wild-type smooth (S/WT), atypical filamentous rough (FR) and atypical multiple-cell-chain (MCR) variants. Exposure of L. monocytogenes cells to sub-lethal acid, salt or heating conditions resulted in similar or increased susceptibility to PUV treatments. Only prior exposure to mild heat stressing significantly enhanced invasion of Caco-2 cells, whereas subjection of L. monocytogenes cells to combined sub-lethal salt, acid and heating conditions produced the greatest reduction in invasiveness. Implications of these findings are discussed. This constitutes the first study to show that pre-exposure to mild conventional food-processing stresses enhances sensitivity of different culture morphotypes of L. monocytogenes to PUV, which is growing in popularity as an alternative or complementary approach for decontamination in the food environment.     

Development of thermal surrogate microorganisms in ground beef for in-plant critical control point validation studies

In search of a suitable surrogate microorganism for in-plant critical control point validation, we compared the rates of thermal inactivation of three bacteria, Enterococcus faecium B2354, Pediococcus parvulus HP, and Pediococcus acidilactici LP, to those of Listeria monocytogenes and Salmonella. Ground beef samples containing 4 and 12% fat were inoculated with E. faecium, L. monocytogenes, and Salmonella Senftenberg 775W and heated at 58, 62, 65, or 68 degrees C. The decimal reduction times (D-values) for E. faecium B2354 in 4 and 12% fat ground beef were 4.4 to 17.7 and 3.6 to 14.6 times greater, respectively, than those for L. monocytogenes or Salmonella Senftenberg 775W at all temperatures tested, with the greatest differences in D-values occurring at 58 and 62 degrees C. Higher fat content protected bacteria from thermal inactivation in general, especially at temperatures lower than 68 degrees C. The heat resistance in a broth medium at 62degrees C of two food-grade bacteria, P. parvulus HP and P. acidilactici LP, was compared with that of the three strains under study. The D-values of P. parvulus HP and P. acidilactici LP were lower than those of E. faecium B2354 but 4.1 and 2.5 times greater, respectively, than those of Salmonella Senftenberg 775W, the most resistant pathogen. These results indicate that thermal treatments of ground beef at 58 to 68 degrees C that kill E. faecium B2354 will also kill Salmonella and L. monocytogenes, and the two Pediococcus isolates may serve as alternate surrogates for validation studies when a less heat-resistant surrogate is desired. However, additional studies in ground beef are needed with the Pediococcus strains in the desired temperature range intended for validation purposes.     

Thermal injury and recovery of Salmonella enterica serovar enteritidis in ground chicken with temperature, pH, and sodium chloride as controlling factors

Cells of Salmonella enterica serovar Enteritidis were grown at 25 and 35 degrees C, heat injured (55, 60, and 62.5 degrees C), and recovered in tryptic soy broth (TSB) at various NaCl concentrations (2.0 and 3.5%) and pH levels (5.5 and 6.5). To assess the interactions of growth temperature, heating temperature, NaCl concentration and pH on the thermal injury and recovery of Salmonella Enteritidis in ground chicken, a randomized design with each experimental combination was used. When a logistic equation for nonlinear survival curves was used, D-values of cells of Salmonella Enteritidis grown at 25 degrees C were 7.60, 5.73, and 4.81 min at 55, 60, and 62.5 degrees C, respectively. For cells grown at 35 degrees C, the D-values were 12.38, 7.45, and 5.70 min at 55, 60, and 62.5 degrees C. The influence of tryptic soy agar and double modified lysine agar (DMLIA) on the recovery of heat-injured cells was determined. Recovery was significantly reduced on DMLIA at increased pH levels and NaCl concentrations. Higher numbers of cells were recovered in TSB with 2.0% NaCl than in TSB with 3.5% NaCl. It was observed that the rate of recovery of heat-injured cells was similar at each pH. Therefore, a pH range of 5.5 to 6.5 does not have a major inhibitory effect on the recovery of Salmonella Enteritidis.     

A predictive model for heat inactivation of Listeria monocytogenes biofilm on stainless steel

Heat treatment of potential biofilm-forming sites is sometimes used for control of Listeria monocytogenes in food processing plants. However, little information is available on the heat treatment required to kill L. monocytogenes present in biofilms. The purpose of this study was to develop a predictive model for the heat inactivation of L. monocytogenes in monoculture biofilms (strains Scott A and 3990) and in biofilms with competing bacteria (Pseudomonas sp. and Pantoea agglomerans) formed on stainless steel in the presence of food-derived soil. Biofilms were produced on stainless steel coupons with diluted tryptic soy broth incubated for 48 h at 25 degrees C. Duplicate biofilm samples were heat treated for 1, 3, 5, and 15 min at 70, 72, 75, 77, and 80 degrees C and tested for survivors using enrichment culture. The experiment was repeated six times. A predictive model was developed using logistic regression analysis of the fraction negative data. Plots showing the probability of L. monocytogenes inactivation in biofilms after heat treatment were generated from the predictive equation. The predictive model revealed that hot water sanitation of stainless steel can be effective for inactivating L. monocytogenes in a biofilm on stainless steel if time and temperature are controlled. For example, to obtain a 75% probability of total inactivation of L. monocytogenes 3990 biofilm, a heat treatment of 80 degrees C for 11.7 min is required. The model provides processors with a risk management tool that provides predicted probabilities of L. monocytogenes inactivation and allows a choice of three heat resistance assumptions. The predictive model was validated using a five-strain cocktail of L. monocytogenes in the presence of food soil.     

A predictive model for the influence of food components on survival of Listeria monocytogenes LM 54004 under high hydrostatic pressure and mild heat conditions

The combination of high hydrostatic pressure with mild temperature was explored to achieve a predictive model of microbial inactivation in food matrix processing. The pressure processing conditions were fixed at 448 MPa for 11 min at the treatment temperature of 41 degrees C, which have been determined as the optimum processing conditions considering six log-cycle reductions of Listeria monocytogenes. Based on the results, response surface methodology (RSM) was performed in the present work, the influence of food components like soybean protein (0-5.00%), sucrose (0.25-13.25%), bean oil (0-10.00%), and pH (4-10) of the food matrix on survival of L. monocytogenes by high pressure and mild heat was studied, and a quadratic predictive model for the influence of food components and pH of food matrix on L. monocytogenes reduction by high pressure and mild heat was built with RSM accurately. The experimental results showed that the efficiency of L. monocytogenes reduction in milk buffer and food matrix designed in the present work, under the HPP treatment process parameters described above, were different. The soybean protein (P=0.0086), sucrose (P<0.0001), and pH (P=0.0136) significantly affected reduction of L. monocytogenes, but the effect of bean oil on reduction of L. monocytogenes was not significant (P=0.1028). The predictive model is significant since the level of significance was P<0.0001 and the calculated F value (11.53) is much greater than the tabulated F value (F(0.01 (14, 5))=9.77). Moreover, the adequacy of the predictive model equation for predicting the level of L. monocytogenes reduction was verified effectively by the validation data.