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The objective of the present study was to analyze the combined effect of heat treatment (55 to 62.5°C) and citral (0 to 3%) on the heat resistance of Escherichia coli O104:H4 inoculated into ground beef. Inoculated meat packages were immersed in a circulating water bath stabilized at 55, 57.5, 60, or 62.5°C for various times. The surviving microbial cells were counted after culture on tryptic soy agar. A factorial design (4 × 4) was used to analyze the effects and interaction of heat treatment and citral. Heat and citral promoted E. coli O104:H4 thermal inactivation, suggesting a synergistic effect. At 55°C, the incorporation of citral at 1, 2, and 3% decreased D-values (control = 42.75 min) by 85, 89, and 91%, respectively (P0.05). A citral concentration-dependent effect (P0.05) also was noted at other evaluated temperatures. These findings could be of value to the food industry for designing a safe thermal process for inactivating E. coli O104:H4 in ground beef under similar thermal inactivation conditions.

The heat resistance of three heat-resistant strains of Salmonella was determined in whole liquid egg (WLE). Inoculated samples in glass capillary tubes were completely immersed in a circulating water bath and held at 56, 58, 60, 62, and 64°C for predetermined lengths of time. The recovery medium was tryptic soy agar with 0.1% sodium pyruvate and 50 ppm of nalidixic acid. Survival data were fitted using log-linear, log-linear with shoulder, and Weibull models using GInaFiT version 1.7. Based on the R2 and mean square error, the log-linear with shoulder and Weibull models consistently produced a better fit to Salmonella survival curves obtained at these temperatures. Contaminated WLE must be heated at 56, 60, and 64°C for at least 33.2, 2.7, and 0.31 min, respectively, to achieve a 4-log reduction of Salmonella; 39.0, 3.1, and 0.34 min, respectively, for a 5-log reduction; and 45.0, 3.5, and 0.39 min, respectively, for a 6-log reduction. The z-values calculated from the D-values were 3.67 and 4.18°C for the log-linear with shoulder and Weibull models, respectively. Thermal death times presented in this study will be beneficial for WLE distributors and regulatory agencies when designing pasteurization processes to effectively eliminate Salmonella in WLE, thereby ensuring the microbiological safety of the product.

Decimal reduction time ( D-value) was calculated for six non-O157 Shiga toxin-producing Escherichia coli (STEC) in a laboratory medium and ground beef. For the laboratory medium, an overnight culture of each strain of STEC was divided into 10-mL sample bags and heated in a water bath for a specific time on the basis of the temperatures. Survival curves were generated by plotting the surviving bacterial population against time, and a linear-log primary model was used to estimate the D-values from survival curves. The z-values (the temperature raised to reduce the D-value by one-tenth) were calculated by plotting the log D-values against temperature. Similarly, for ground beef, six fat contents, 5, 10, 15, 20, 25, and 30% of ground beef were formulated for this study. Inoculated meat was divided into 5-g pouches and submerged in a water bath set at specific temperatures (55, 60, 65, 68, and 71.1°C). The average D-value for these strains in a laboratory medium was 17.96 min at 55°C, which reduced significantly ( P0.05) to 1.58 min at 60°C, and then further reduced ( P0.05) to 0.46 min at 65°C. In ground beef, a negative correlation ( P0.05) between fat content of ground beef and D-values was observed at 55°C. However, at temperatures greater than 60°C, there was no impact ( P0.05) of fat content of ground beef on the thermal resistance of non-O157 STECs. Irrespective of the fat content of ground beef, the D-values ranged from 15.93 to 11.69, 1.15 to 1.12, and 0.14 to 0.09 min and 0.05 at 55, 60, 65, and 68°C, respectively. The data generated from this study can be helpful for the meat industry to develop predictive models for thermal inactivation of non-O157 STECs in ground beef with varying fat content.

Kinetic growth data for Bacillus cereus grown from spores were collected in cooked beans under several isothermal conditions (10 to 49°C). Samples were inoculated with approximately 2 log CFU/g heat-shocked (80°C for 10 min) spores and stored at isothermal temperatures. B. cereus populations were determined at appropriate intervals by plating on mannitol-egg yolk-polymyxin agar and incubating at 30°C for 24 h. Data were fitted into Baranyi, Huang, modified Gompertz, and three-phase linear primary growth models. All four models were fitted to the experimental growth data collected at 13 to 46°C. Performances of these models were evaluated based on accuracy and bias factors, the coefficient of determination ( R2), and the root mean square error. Based on these criteria, the Baranyi model best described the growth data, followed by the Huang, modified Gompertz, and three-phase linear models. The maximum growth rates of each primary model were fitted as a function of temperature using the modified Ratkowsky model. The high R2 values (0.95 to 0.98) indicate that the modified Ratkowsky model can be used to describe the effect of temperature on the growth rates for all four primary models. The acceptable prediction zone (APZ) approach also was used for validation of the model with observed data collected during single and two-step dynamic cooling temperature protocols. When the predictions using the Baranyi model were compared with the observed data using the APZ analysis, all 24 observations for the exponential single rate cooling were within the APZ, which was set between -0.5 and 1 log CFU/g; 26 of 28 predictions for the two-step cooling profiles also were within the APZ limits. The developed dynamic model can be used to predict potential B. cereus growth from spores in beans under various temperature conditions or during extended chilling of cooked beans.

The heat resistance of three heat-resistant strains of

The inactivation of E. coli O157:H7 in ground beef patties cooked in a skillet was investigated. Ground beef patties inoculated with a mixture of five strains of E. coli O157:H7 were cooked in a Farberware skillet set at a temperature of 275°F (137°C). Eight type K thermocouples connected to a data logger were used to record the temperatures at eight points within the patty. The cooking times studied ranged from 2.25 min to 4 min. Tryptic soy agar plates overlaid with sorbitol MacConkey agar were used for recovery of E. coli O157:H7. Heating of ground beef patties to an internal temperature endpoint of 155°F (68.3°C) resulted in 4-log cycle reductions of the organism. The results of this investigation conducted under conditions simulating those that occur in the retail food industry provide a basis for ensuring safety against E. coli O157:H7 in ground beef patties.

The heat resistance of nonproteolytic type B and type E Clostridium botulinum spores in phosphate buffer and turkey slurry was determined from 70 to 90°C. Thermal-death times were determined in vials heated using a water bath. Recovery of heat-injured spores was on reinforced clostridial medium (RCM) and tryptic soy agar (TSA) with and without added lysozyme (10 μg/ml). Decimal-reduction times (D-values) were determined by fitting a survival model to the data using a curve-fitting program. The apparent or measured heat resistance was maximum with RCM supplemented with lysozyme. The D-values at 80°C for type E spores in buffer ranged from 1.03 min for strain Whitefish to 4.51 min for strain Saratoga. The D-value for the most heat-resistant nonproteolytic type B strain KAP B5 in buffer was 4.31 min at 80°C. The z-values in buffer for all strains were very similar, ranging from 8.35 to 10.08°C.Turkey slurry offered protection to the spores with a concomitant increase in heat resistance. The D-values in turkey slurry ranged from 51.89 min at 70°C to 1.18min at 85°C for type E strain Alaska (z = 9.90°C) and from 32.53 min at 75°C to 0.80 min at 90°C for nonproteolytic type B strain KAP B5 (z = 9.43°C). Thermal-death-time values from this study will assist food processors to design thermal processes that ensure safety against nonproteolytic C. botulinum in cook/chill foods.

Demand for minimally processed refrigerated foods with reduced salt levels has stimulated renewed interest in the potential for survival and growth of psychrotrophic, nonproteolytic Clostridium botulinum type B spores. As part of a project to better define food-processing requirements, the heat resistance (75 to 90°C) of nonproteolytic C. botulinum type B spores was assessed in turkey containing 1 to 3% (wt/vol) salt (sodium chloride). Heated spores were recovered both on reinforced clostridial medium (RCM) with lysozyme and on RCM having the same salt levels as the heating menstruum. When the recovery medium contained no salt, D-values in turkey slurry containing 1% salt were 42.1, 17.1, 7.8, and 1.1 min at 75, 80, 85, and 90°C, respectively. Increasing levels (2 and 3%, wt/vol) of salt in the turkey slurry reduced the heat resistance as evidenced by reduced spore D-values. Also, apparent or measured heat resistance was decreased with increasing salt concentration in the heating menstruum and the recovery medium. The z-values in turkey slurry for all treatments were similar, ranging from 8.47 to 10.08°C. These data will assist food processors to design thermal processes that ensure safety against nonproteolytic C. botulinum type B spores in cook/chill foods while minimizing quality losses.

A polymerase chain reaction (PCR) procedure was developed for direct detection of Clostridium perfringens strains with potential for food poisoning in raw beef samples. An oligonucleotide primer pair was used to amplify a 364 base pair sequence internal to the C. perfringens enterotoxin gene. One milliliter portions of the meat homogenates were inoculated into cooked meat medium (CMM) or reduced Fluid Thioglycollate (FTG) medium and incubated at 37°C. Portions sampled at 2, 4, 6, 8 and 24 h of enrichment were assayed for detection of the enterotoxin sequence by PCR. Amplification of the 364 bp sequence could be detected in 6 h by agarose gel electrophoresis and as early as 2 h by hybridization to a 150 bp digoxigenin (DIG)-labeled probe. To increase the sensitivity of the detection assay a commercial chromosomal deoxyribonucleic acid (DNA) extraction assay was compared with a nested PCR approach. Both methods allowed detection of less than 1 log10 colony forming units (CFU)/g of C. perfringens strains harboring the enterotoxin gene, with no interference with the background microflora present in the raw ground beef.

The ability of Clostridium perfringens spores to germinate and grow was studied to determine a safe cooling rate for cooked beef. Beef samples were inoculated with a cocktail of three strains of heat-shocked C. perfringens spores (NCTC 8238, NCTC 8239 and ATCC 10288), vacuum-packaged, and cooked in a stirred water bath to an internal temperature of 60°C in 1 h. Then, samples were cooled through the temperature range of 54.4°C to 7.2°C at rates varying from 6 to 18 h. Samples were removed at various times during cooling to determine if the spores had germinated and multiplied. The samples were plated on tryptose-sulfite-cycloserine agar and incubated anaerobically at 37°C for 48 h. Minimal growth was observed with cooling periods of up to 15 h. However, with the time to achieve 7.2°C extended to 18 h, C. perfringens spores germinated and grew from an inoculum of approximately 1.5 log10 to about 6.0 log10 CFU/g. This study indicated that pasteurized cooked beef must be cooled to 72°C in 15 h or less to prevent C. perfringens foodborne disease outbreaks.

The objective of this study was to assess the ability of Bacillus cereus spores to germinate and grow in order to determine a safe cooling rate for cooked rice, beans, and pasta, rice-chicken (4:1), rice-chicken-vegetables (3:1:1), rice-beef (4:1), and rice-beef-vegetables (3:1:1). Samples were inoculated with a cocktail of four strains of heat-shocked (80°C for 10 min) B. cereus spores (NCTC 11143, 935A/74, Brad 1, and Mac 1) to obtain a final spore concentration of approximately 2 log CFU/g. Thereafter, samples were exponentially cooled through the temperature range of 54.5 to 7.2°C in 6, 9, 12, 15, 18, and 21 h. At the end of the cooling period, samples were removed and plated on mannitol egg yolk polymyxin agar. The plates were incubated at 30°C for 24 h. The net B. cereus growth from spores in beans was

Heat-resistant spores of Clostridium perfringens may germinate and multiply in cooked meat and poultry products when the rate and extent of cooling does not occur in a timely manner. Therefore, six cooling models (PMP 7.0 broth model; PMIP uncured beef, chicken, and pork models; Smith-Schaffner version 3; and UK IFR ComBase Perfringens Predictor) were evaluated for relative performance in predicting growth of C. perfringens under dynamic temperature conditions encountered during cooling of cooked, uncured meat and poultry products. The predicted growth responses from the models were extensively compared with those observed in food. Data from 188 time-temperature cooling profiles (176 for single-rate exponential cooling and 12 for dual-rate exponential cooling) were collected from 17 independent sources (16 peer-reviewed publications and one report) for model evaluation. Data were obtained for a variety of cooked products, including meat and poultry slurries, ground meat and poultry products with and witho...

The surfaces of ready-to-eat meats are susceptible to postprocessing contamination by Listeria monocytogenes. This study quantified the lag-phase durations (LPD) and growth rates (GR) of L. monocytogenes on the surfaces of cooked ham as affected by sorbate solutions of different concentrations and pH levels. Slices of cooked ham inoculated with a four-strain mixture of L. monocytogenes (ca. 10(3) CFU/g) were surface treated with sorbate solutions of 0 to 4% (wt/vol) at pH 4.0 to 6.5, vacuum packaged, and stored at 4 to 12 °C for up to 45 days. The LPD and GR of L. monocytogenes were used to develop response surface models. The models estimated that the LPD of L. monocytogenes in samples treated with solutions of pH 4.0 to 5.5 (no sorbate) were 0 to 11 days and the GR were 0.25 to 0.36 log CFU/day, respectively, at 4 °C. With the treatments of 2 and 4% (wt/vol) sorbate solutions, the LPD were estimated to be extended to 2 to 26 days and 34 to45 days, and the GR were reduced to 0.15 to 0.30 and 0 to 0.19 log CFU/day, respectively. At 4 °C, increasing sorbate concentrations by 1% (wt/vol) to 2, 3, and 4% (wt/vol) at pH 5.5 to 4.0 led to an extension of LPD by 2 to 11, 10 to 19, and 18 to 27 days, whereas the GR were reduced by 0.037 to 0.055, 0.048 to 0.066, and 0.060 to 0.078 log CFU/day, respectively. Sorbate also extended the LPD and reduced the GR of L. monocytogenes at 8 and 12 °C. Results indicated that sorbate concentration and pH level were significant factors affecting the LPD and GR of L. monocytogenes and that the combination of sorbate and low pH has potential for use as a surface treatment to control L. monocytogenes on meat surfaces.

The interactive effects of heating temperature (55 to 65°C), sodium chloride (NaCl; 0 to 2%), and green tea 60% polyphenol extract (GTPE; 0 to 3%) on the heat resistance of a five-strain mixture of Listeria monocytogenes in ground turkey were determined. Thermal death times were quantified in bags that were submerged in a circulating water bath set at 55, 57, 60, 63, and 65°C. The recovery medium was tryptic soy agar supplemented with 0.6% yeast extract and 1% sodium pyruvate. D-values were analyzed by second-order response surface regression for temperature, NaCl, and GTPE. The data indicated that all three factors interacted to affect the inactivation of the pathogen. The D-values for turkey with no NaCl or GTPE at 55, 57, 60, 63, and 65°C were 36.3, 20.8, 13.2, 4.1, and 2.9 min, respectively. Although NaCl exhibited a concentration-dependent protective effect against heat lethality on L. monocytogenes in turkey, addition of GTPE rendered the pathogen more sensitive to the lethal effect of heat. GTPE levels up to 1.5% interacted with NaCl and reduced the protective effect of NaCl on heat resistance of the pathogen. Food processors can use the predictive model to design an appropriate heat treatment that would inactivate L. monocytogenes in cooked turkey products without adversely affecting the quality of the product.

The risk of non-O157 Shiga toxin–producing Escherichia coli strains has become a growing public health concern. Several studies characterized the behavior of E. coli O157:H7; however, no reports on the influence of multiple factors on E. coli O104:H4 are available. This study examined the effects and interactions of temperature (7 to 46°C), pH (4.5 to 8.5), and water activity (aw; 0.95 to 0.99) on the growth kinetics of E. coli O104:H4 and developed predictive models to estimate its growth potential in foods. Growth kinetics studies for each of the 23 variable combinations from a central composite design were performed. Growth data were used to obtain the lag phase duration (LPD), exponential growth rate, generation time, and maximum population density (MPD). These growth parameters as a function of temperature, pH, and aw as controlling factors were analyzed to generate second-order response surface models. The results indicate that the observed MPD was dependent on the pH, aw, and temperature of the gro...

Isothermal inactivation studies are commonly used to quantify thermal inactivation kinetics of bacteria. Meta-analyses and comparisons utilizing results from multiple sources have revealed large variations in reported thermal resistance parameters for Salmonella, even when in similar food materials. Different laboratory or regression methodologies likely are the source of methodology-specific artifacts influencing the estimated parameters; however, such effects have not been quantified. The objective of this study was to evaluate the effects of laboratory and regression methodologies on thermal inactivation data generation, interpretation, modeling, and inherent error, based on data generated in two independent laboratories. The overall experimental design consisted of a cross-laboratory comparison using two independent laboratories (Michigan State University and U.S. Department of Agriculture, Agricultural Research Service, Eastern Regional Research Center [ERRC] laboratories), both conducting isothermal Salmonella inactivation studies (55, 60, 62°C) in ground beef, and each using two methodologies reported in prior studies. Two primary models (log-linear and Weibull) with one secondary model (Bigelow) were fitted to the resultant data using three regression methodologies (two two-step regressions and a one-step regression). Results indicated that laboratory methodology impacted the estimated D60°C- and z-values (α = 0.05), with the ERRC methodology yielding parameter estimates ∼25% larger than the Michigan State University methodology, regardless of the laboratory. Regression methodology also impacted the model and parameter error estimates. Two-step regressions yielded root mean square error values on average 40% larger than the one-step regressions. The Akaike Information Criterion indicated the Weibull as the more correct model in most cases; however, caution should be used to confirm model robustness in application to real-world data. Overall, the results suggested that laboratory and regression methodologies have a large influence on resultant data and the subsequent estimation of thermal resistance parameters.

D- and z-values for Listeria monocytogenes were obtained for two Mexican meat entrées: pork meat marinated in tomatillo (green tomato) sauce (PTS) and beef marinated in a red chili sauce (BRCS), with addition of 0, 200, and 800 ppm of grapefruit seed extract (GSE). Meat samples inoculated with L. monocytogenes were packaged in sterile bags, immersed in a water bath, and held at 55, 57.5, 60, and 62.5°C for different periods of time. Depending upon the temperature, D-values at 0 ppm of GSE ranged from 26.19 to 2.03 min in BRCS and 26.41 to 0.8 min in PTS. Adding 800 ppm of GSE to BRCS thermally treated at 55 and 62.5°C significantly decreased inactivation time by 35%. A reduction in time of 25.9, 10.6, and 40.1% at 55, 57.5, and 60°C, respectively, was observed in PTS with 800 ppm of GSE. The z-values of L. monocytogenes were not significantly affected by GSE addition; average z-values were 7.25 and 5.09°C for BRCS and PTS, respectively. Estimated thermal lethality for a 7-D log reduction of L. monocytogenes under commercial-size sous-vide conditions at a reference temperature of 55°C was reached at 78 and 71 min for BRCS without and with 800 ppm of GSE, respectively. For PTS, 7-D reduction was attained at 69 and 61 min without and with addition of 800 ppm of GSE, respectively. Supplementing both Mexican meat entrées (BRCS and PTS) with 800 ppm of GSE rendered L. monocytogenes cells more sensitive to the lethal effect of heat. The results of this study will assist the retail food industry in designing acceptance limits on critical control points pertaining to cooking regimes to effectively eliminate L. monocytogenes in BRCS and PTS sous-vide processed Mexican meat entrées.

We investigated the heat resistance of an eight-strain cocktail of Salmonella serovars in chicken supplemented with trans cinnamaldehyde (0 to 1.0%, wt/wt) and carvacrol (0 to 1.0%, wt/wt). Inoculated meat was packaged in bags that were completely immersed in a circulating water bath and held at 55 to 71°C for predetermined lengths of time. The recovery medium was tryptic soy agar supplemented with 0.6% yeast extract and 1% sodium pyruvate. D-values in chicken, determined by linear regression, were 17.45, 2.89, 0.75, and 0.29 min at 55, 60, 65, and 71°C, respectively (z = 9.02°C). Using a survival model for nonlinear survival curves, D-values in chicken ranged from 13.52 min (D(1), major population) and 51.99 min (D(2), heat-resistant subpopulation) at 55°C to 0.15 min (D(1)) and 1.49 min (D(2)) at 71°C. When the Salmonella cocktail was in chicken supplemented with 0.1 to 1.0% trans-cinnamaldehyde or carvacrol, D-values calculated by both approaches were consistently less at all temperatures. This observation suggests that the addition of natural antimicrobials to chicken renders Salmonella serovars more sensitive to the lethal effect of heat. Thermal death times from this study will be beneficial to the food industry in designing hazard analysis and critical control point plans to effectively eliminate Salmonella contamination in chicken products used in this study.

Interest in chitosan, a biodegradable, nontoxic, non-antigenic, and biocompatible biopolymer isolated from shellfish, arises from the fact that chitosans are reported to exhibit numerous health-related beneficial effects, including strong antimicrobial and antioxidative activities in foods. The extraordinary interest in the chemistry and application in agriculture, horticulture, environmental science, industry, microbiology, and medicine is attested by about 17,000 citations on this subject in the Scopus database. A special need exists to develop a better understanding of the role of chitosans in ameliorating foodborne illness. To contribute to this effort, this overview surveys and interprets our present knowledge of the chemistry and antimicrobial activities of chitosan in solution, as powders, and in edible films and coating against foodborne pathogens, spoilage bacteria, and pathogenic viruses and fungi in several food categories. These include produce, fruit juices, eggs and dairy, cereal, meat, and seafood products. Also covered are antimicrobial activities of chemically modified and nanochitosans, therapeutic properties, and possible mechanisms of the antimicrobial, antioxidative, and metal chelating effects. Further research is suggested in each of these categories. The widely scattered data on the multifaceted aspects of chitosan microbiology, summarized in the text and in 10 tables and 8 representative figures, suggest that low-molecular-weight chitosans at a pH below 6.0 presents optimal conditions for achieving desirable antimicrobial and antioxidative-preservative effects in liquid and solid foods. We are very hopeful that the described findings will be a valuable record and resource for further progress to improve microbial food safety and food quality.

Inhibition of Clostridium perfringens spore germination and outgrowth in ground turkey roast containing minimal ingredients (salt and sugar), by buffered vinegar (MOstatin V) and a blend (buffered) of lemon juice concentrate and vinegar (MOstatin LV) was evaluated. Ground turkey roast was formulated to contain sea salt (1.5%), turbinado sugar (0.5%), and various concentrations of MOstatin V (0.75, 1.25, or 2.5%) or MOstatin LV (1.5, 2.5, or 3.5%), along with a control (without MOstatins). The product was inoculated with a three-strain spore cocktail of C. perfringens to obtain initial spore levels of ca. 2.0 to 0.5 log CFU/g. Inoculated products were vacuum packaged, heat shocked for 20 min at 75 degrees C, and cooled exponentially from 54.4 to 4.0 degrees C in 6.5, 9, 12, 15, 18, or 21 h. In control samples without MOstatin V or MOstatin LV, C. perfringens populations reached 2.98, 4.50, 5.78, 7.05, 7.88, and 8.19 log CFU/g (corresponding increases of 0.51, 2.29, 3.51, 4.79, 5.55, and 5.93 log CFU/g) in 6.5, 9, 12, 15, 18, and 21 h of chilling, respectively. MOstatin V (2.5%) and MOstatin LV (3.5%) were effective in inhibiting C. perfringens spore germination and outgrowth in ground turkey roast to

We investigated the heat resistance of a four-strain mixture of Escherichia coli O157:H7 in raw ground beef in both the absence and presence of white and green tea powders and an apple skin extract. Inoculated meat was cooked using the sous-vide technique, i.e., the meat was packaged in sterile bags and completely immersed in a circulating water bath at low temperature for a period of time. The bags were cooked for 1 h to an internal temperature of 55, 58, 60, or 62.5 degrees C, and then held from 240 min at 55 degrees C to 10 min at 62.5 degrees C. The surviving bacteria were enumerated by spiral plating onto tryptic soy agar overlaid with sorbitol-MacConkey agar. Inactivation kinetics of the pathogens deviated from first-order kinetics. D-values (time, in minutes, required for the bacteria to decrease by 90%) in the control beef ranged from 67.79 min at 55 degrees C to 2.01 min at 62.5 degrees C. D-values determined by a logistic model ranged from 36.22 (D1, the D-value of a major population of surviving cells) and 112.79 (D2, the D-value of a minor subpopulation) at 55 degrees C to 1.39 (D1) and 3.00 (D2) at 62.5 degrees C. A significant increase (P0.05) in the sensitivity of the bacteria to heat was observed with the addition of 3% added antimicrobials. D-value reductions of 62 to 74% were observed with apple powder and 18 to 58% with tea powders. Thermal death times from this study will assist the retail food industry to design cooking regimes that ensure the safety of beef contaminated with E. coli O157:H7.

Irradiated ground beef samples (ca. 3-g portions with ca. 25% fat) inoculated with Yersina pestis strain KIM5 (ca. 6.7 log CFU/g) were heated in a circulating water bath stabilized at 48.9, 50, 52.5, 55, 57.5, or 60 degrees C (120, 122, 126.5, 131, 135.5, and 140 degrees F, respectively). Average D-values were 192.17, 34.38, 17.11, 3.87, 1.32, and 0.56 min, respectively, with a corresponding z-value of 4.67 degrees C (8.41 degrees F). In related experiments, irradiated ground beef patties (ca. 95 g per patty with ca. 25% fat) were inoculated with Y. pestis strains KIMS or CDC-A1122 (ca. 6.0 log CFU/g) and cooked on an open-flame gas grill or on a clam-shell type electric grill to internal target temperatures of 48.9, 60, and 71.1 degrees C (120, 140, and 160 degrees F, respectively). For patties cooked on the gas grill, strain KIM5 populations decreased from ca. 6.24 to 4.32, 3.51, and < or = 0.7 log CFU/g at 48.9, 60, and 71.1 degrees C, respectively, and strain CDC-A1122 populations decreased to 3.46 log CFU/g at 48.9 degrees C and to < or = 0.7 log CFU/g at both 60 and 71.1 degrees C. For patties cooked on the clam-shell grill, strain KIM5 populations decreased from ca. 5.96 to 2.53 log CFU/g at 48.9 degrees C and to < or = 0.7 log CFU/g at 60 or 71.1 degrees C, and strain CDC-A1122 populations decreased from ca. 5.98 to < or = 0.7 log CFU/g at all three cooking temperatures. These data confirm that cooking ground beef on an open-flame gas grill or on a clam-shell type electric grill to the temperatures and times recommended by the U.S. Department of Agriculture and the U.S. Food and Drug Administration Food Code, appreciably lessens the likelihood, severity, and/or magnitude of consumer illness if the ground beef were purposefully contaminated even with relatively high levels of Y. pestis.

Heat-resistant spores of Clostridium perfringens may germinate and multiply in cooked meat and poultry products when the rate and extent of cooling does not occur in a timely manner. Therefore, six cooling models (PMP 7.0 broth model; PMIP uncured beef, chicken, and pork models; Smith-Schaffner version 3; and UK IFR ComBase Perfringens Predictor) were evaluated for relative performance in predicting growth of C. perfringens under dynamic temperature conditions encountered during cooling of cooked, uncured meat and poultry products. The predicted growth responses from the models were extensively compared with those observed in food. Data from 188 time-temperature cooling profiles (176 for single-rate exponential cooling and 12 for dual-rate exponential cooling) were collected from 17 independent sources (16 peer-reviewed publications and one report) for model evaluation. Data were obtained for a variety of cooked products, including meat and poultry slurries, ground meat and poultry products with and without added ingredients (e.g., potato starch, sodium triphosphate, and potassium tetrapyrophosphate), and processed products such as ham and roast beef. Performance of the models was evaluated using three sets of criteria, and accuracy was defined within a 1- to 2-log range. The percentages of accurate, fail-safe, or fail-dangerous predictions for each cooling model differed depending on which criterion was used to evaluate the data set. Nevertheless, the combined percentages of accurate and fail-safe predictions based on the three performance criteria were 34.66 to 42.61% for the PMP 7.0 beef broth model, 100% for the PMIP cooling models for uncured beef, uncured pork and uncured chicken, 80.11 to 93.18% for the Smith-Schaffner cooling model, and 74.43 to 85.23% for the UK IFR ComBase Perfringens Predictor model during single-rate exponential chilling. Except for the PMP 7.0 broth model, the other five cooling models (PMIP, Smith-Schaffner, and UK IFR ComBase) are useful and reliable tools that food processors and regulatory agencies can use to evaluate the safety of cooked or heat-treated uncured meat and poultry products exposed to cooling deviations or to develop customized cooling schedules.

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.

Inhibition of Clostridium perfringens spore germination and outgrowth by carvacrol, cinnamaldehyde, thymol, and oregano oil was evaluated during abusive chilling of cooked ground beef (75% lean) obtained from a local grocery store. Test substances were mixed into thawed ground beef at concentrations of 0.1, 0.5, 1.0, or 2.0% (wt/wt) along with a heat-activated three-strain C. perfringens spore cocktail to obtain final spore concentrations of ca. 2.8 log spores per g. Aliquots (5 g) of the ground beef mixtures were vacuum-packaged and then cooked in a water bath, the temperature of which was raised to 60 degrees C in 1 h. The products were cooled from 54.4 to 7.2 degrees C in 12, 15, 18, or 21 h, resulting in 3.18, 4.64, 4.76, and 5.04 log CFU/ g increases, respectively, in C. perfringens populations. Incorporation of test compounds (> or = 0.1%) into the beef completely inhibited C. perfringens spore germination and outgrowth (P < or = 0.05) during exponential cooling of the cooked beef in 12 h. Longer chilling times (15, 18, and 21 h) required greater concentrations to inhibit spore germination and outgrowth. Cinnamaldehyde was significantly (P < 0.05) more effective (< 1.0 log CFU/g growth) at a lower concentration (0.5%) at the most abusive chilling rate evaluated (21 h) than the other compounds. Incorporation of lower levels of these test compounds with other antimicrobials used in meat product formulations may reduce the potential risk of C. perfringens germination and outgrowth during abusive cooling regimes.

Ionizing radiation can be effective in controlling the growth of food spoilage and foodborne pathogenic bacteria. This study reports on an investigation of the effectiveness of irradiation treatment to eliminate Listeria monocytogenes on laboratory-inoculated broccoli, cabbage, tomatoes, and mung bean sprouts. Irradiation of broccoli and mung bean sprouts at 1.0 kGy resulted in reductions of approximately 4.88 and 4.57 log CFU/g, respectively, of a five-strain cocktail of L. monocytogenes. Reductions of approximately 5.25 and 4.14 log CFU/g were found with cabbage and tomato, respectively, at a similar dose. The appearance, color, texture, taste, and overall acceptability did not undergo significant changes after 7 days of postirradiation storage at 4 degrees C, in comparison with control samples. Therefore, low-dose ionizing radiation treatment could be an effective method for eliminating L. monocytogenes on fresh and fresh-cut produce.

This study evaluated the effect of organic acids and spices, alone or combined, on Clostridium perfringens growth in cooked ground beef during alternative cooling procedures. Ground beef was inoculated with a three-strain cocktail of C. perfringens (ATCC 10388, NCTC 8238, and NCTC 8239) at 2 log spores per g and prepared following an industrial recipe (10% water, 1.5% sodium chloride, and 0.5% sodium triphosphate [wt/wt]). Treatments consisted of the base meat plus combinations of commercial solutions of sodium lactate or sodium citrate (0 or 2%, wt/wt) with chili, garlic and herbs, curry, oregano, or clove in commercial powder form (0 or 1%, wt/wt). Untreated meat was used as a control. Vacuum-packaged samples of each treatment were cooked (75 degrees C for 20 min) and cooled from 54.4 to 7.2 degrees C in 15, 18, or 21 h. Spore counts were estimated after inoculation, cooking, and cooling. All treatments containing sodium citrate reduced the population of C. perfringens about 0.38 to 1.14 log units during each of the three cooling procedures. No sodium citrate and spice treatment combinations showed antagonisms or synergisms. Regardless of the cooling time, the control ground beef or treatments with any of the five spices alone supported C. perfringens growth above the U.S. Department of Agriculture stabilization guidelines of 1 log unit. Except for the 21-h cooling period, addition of sodium lactate prevented C. perfringens growth over 1 log unit. Depending on the cooling time and spice, some combinations of sodium lactate and spice kept C. perfringens growth below 1 log unit.

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.

Inhibition of the germination and outgrowth of Clostridium perfringens by buffered sodium citrate (Ional) and buffered sodium citrate supplemented with sodium diacetate (Ional Plus) during the abusive chilling of roast beef and injected pork was evaluated. Beef top rounds or pork loins were injected with a brine containing NaCl, potato starch, and potassium tetrapyrophosphate to yield final in-product concentrations of 0.85, 0.25, and 0.20%, respectively. Products were ground and mixed with Ional or Ional Plus at 0, 0.5, 1.0, and 2.0%. Each product was mixed with a three-strain C. perfringens spore cocktail to obtain final spore concentrations of ca. 2.5 log10 spores per g. Chilling of roast beef from 54.4 to 7.2 degrees C resulted in C. perfringens population increases of 1.51 and 5.27 log10 CFU/g for 18- and 21-h exponential chill rates, respectively, while chilling of injected pork resulted in increases of 3.70 and 4.41 log10 CFU/g. The incorporation of Ional into the roast beef formulation resulted in C. perfringens population reductions of 0.98, 1.87, and 2.47 log10 CFU/g with 0.5, 1.0, and 2.0% Ional, respectively, over 18 h of chilling, while > or = 1.0% Ional Plus was required to achieve similar reductions (reductions of 0.91 and 2.07 log10 CFU/g were obtained with 1.0 and 2.0% Ional Plus, respectively). An Ional or Ional Plus concentration of > or = 1.0% was required to reduce C. perfringens populations in roast beef or injected pork chilled from 54.4 to 7.2 degrees C in 21 h. Cooling times for roast beef or injected pork products after heat processing can be extended to 21 h through the incorporation of > or = 1.0% Ional or Ional Plus into the formulation to reduce the potential risk of C. perfringens germination and outgrowth.

The risk of non-O157 Shiga toxin-producing Escherichia coli strains has become a growing public health concern. Several studies characterized the behavior of E. coli O157:H7; however, no reports on the influence of multiple factors on E. coli O104:H4 are available. This study examined the effects and interactions of temperature (7 to 46°C), pH (4.5 to 8.5), and water activity (aw ; 0.95 to 0.99) on the growth kinetics of E. coli O104:H4 and developed predictive models to estimate its growth potential in foods. Growth kinetics studies for each of the 23 variable combinations from a central composite design were performed. Growth data were used to obtain the lag phase duration (LPD), exponential growth rate, generation time, and maximum population density (MPD). These growth parameters as a function of temperature, pH, and aw as controlling factors were analyzed to generate second-order response surface models. The results indicate that the observed MPD was dependent on the pH, aw, and temperature of the growth medium. Increasing temperature resulted in a concomitant decrease in LPD. Regression analysis suggests that temperature, pH, and aw significantly affect the LPD, exponential growth rate, generation time, and MPD of E. coli O104:H4. A comparison between the observed values and those of E. coli O157:H7 predictions obtained by using the U. S. Department of Agriculture Pathogen Modeling Program indicated that E. coli O104:H4 grows faster than E. coli O157:H7. The developed models were validated with alfalfa and broccoli sprouts. These models will provide risk assessors and food safety managers a rapid means of estimating the likelihood that the pathogen, if present, would grow in response to the interaction of the three variables assessed.

Vegetative cells of Clostridium perfringens enterotoxigenic strains NCTC 8679, NCTC 8238. and H6 were grown at 37 degrees C followed by a 60-min exposure to 28 degrees C or 46 degrees C. D10-values, as a measure of thermal resistance at 60 degrees C, were significantly lower for 28 degrees C exposures as compared with cultures given 37 and 46 degrees C exposures. Following refrigeration at 4 degrees C for 24 h, D10-values for the 37 and 46 degrees C samples could not be differentiated from 28 degrees C samples. Western immunoblot analyses of lysates from heat-adapted cells also detected the increased expression of proteins reacting with antiserum directed against the molecular chaperonins from Escherichia coli; GroEL, DnaJ, and the small acid soluble protein from Bacillus subtilis, SspC. Differential scanning calorimetry (DSC) identified thermal transitions corresponding to ribosomal protein denaturations at 72.1 +/- 0.5 degrees C. Any cellular heat adaptations in the DSC profiles were lost following refrigeration for several days to simulate minimally processed food storage conditions. Further analyses of high-speed pellets from crude cell extract fractions using two-dimensional gel electrophoresis detected the differential gene expression of at least four major proteins in heat-adapted vegetative cells of C. perfringens. N-terminal amino acid analyses identified two of the proteins as glyceraldehyde 3-phosphate dehydrogenase and rubrerythrin. Both appear to have roles in this anaerobe under stressful conditions.

The effects and interactions of heating temperature (55 to 65 degrees C), pH (4 to 8), salt (NaCl; 0 to 6%, wt/vol), and sodium pyrophosphate (SPP; 0 to 0.3%, wt/vol) on the heat inactivation of a four-strain mixture of Listeria monocytogenes in beef gravy were examined. A factorial experimental design comparing 48 combinations of heating temperature, salt concentration, pH value, and SPP content was used. Heating was carried out using a submerged-coil heating apparatus. The recovery medium was plate count 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, pH, NaCl, and SPP levels. Whereas increasing the NaCl concentration protected L. monocytogenes against the lethal effect of heat, high SPP concentrations increased heat sensitivity. Also, low pH values increased heat sensitivity of L. monocytogenes. The four variables interacted to affect the inactivation of the pathogen. Thermal resistance of L. monocytogenes can be lowered by combining these intrinsic factors. A predictive model that described the combined effect of temperature, pH, NaCl, and SPP levels on thermal resistance of L. monocytogenes was developed. The model can predict D-values for any combination of temperature, pH, NaCl, and SPP that are within the range of those tested. Using this predictive model, food processors should be able to design adequate thermal regimes to eliminate L. monocytogenes in thermally processed foods.

The ability of 16 foodborne pathogens, representative of 5 different species, to grow during cooling of previously sterilized cooked beef was studied to determine a safe cooling rate. Auto-claved ground beef samples (3 g) were inoculated with heat-shocked spores of Bacillus cereus (strain BH 86) or Clostridium botulinum (nonproteolytic type B strains CBW 25, 17B, and KAP B5 and type E strains Whitefish, Saratoga, and Alaska) or vegetative cells of Listeria monocytogenes (strains HO-VJ-S, V-7, and Scott A), Staphylococcus aureus (strains 196E, B121, and B 124), or Salmonella serotypes ( S. dublin , S. enteritidis , and S. typhimurium ), vacuum-packaged, and cooked in a stirred water bath to an internal temperature of 60°C in I h. In some experiments combinations of C. botulinum and B. cereus spores or S. aureus and salmonellae vegetative cells were used. Heated samples were cooled through the temperature range of 54.4 to 7.2°C at rates varying from 6 to 21 h. Samples were removed at various times during cooling to determine if growth of the pathogens had occurred. No growth was observed with cooling periods of up to 21h. This study with the model meat system (3 g autoclaved ground beef inoculated with selected pathogens and then pasteurized) indicated that cooling from 52.4 to 7.2°C in up to 21 h would not pose a food safety hazard from growth of these pathogens.