A predictive growth model for Clostridium botulinum during cooling of cooked uncured ground beef.

A dynamic model to predict the germination and outgrowth of Clostridium botulinum spores in cooked ground beef was presented. Raw ground beef was inoculated with a ten-strain C. botulinum spore cocktail to achieve approximately 2 log spores/g. The inoculated ground beef was vacuum packaged, cooked to 71 °C to heat shock the spores, cooled to below 10 °C, and incubated isothermally at temperatures from 10 to 46 °C. C. botulinum growth was quantified and fitted into the primary Baranyi Model. Secondary models were fitted to maximum specific growth rate and lag phase duration using Modified Ratkowsky equation (R2 0.96) and hyperbolic function (R2 0.94), respectively. Similar experiments were also performed under non-isothermal (cooling) conditions. Acceptable zone prediction (APZ) analysis was conducted on growth data collected over 3 linear cooling regimes from the current study. The model performance (prediction errors) for all 22 validation data points collected in the current work were within the APZ limits (-1.0 to +0.5 log CFU/g). Additionally, two other growth data sets of C. botulinum reported in the literature were also subjected to the APZ analysis. In these validations, 20/22 and 10/14 predictions fell within the APZ limits. The model presented in this work can be employed to predict C. botulinum spore germination and growth in cooked uncured beef under non-isothermal conditions. The beef industry processors and food service organizations can utilize this predictive microbial model for cooling deviations and temperature abused situations and in developing customized process schedules for cooked, uncured beef products.

Occurrence of Listeria monocytogenes in Ready-to-Eat Meat and Poultry Product Verification Testing Samples from U.S. Department of Agriculture-Regulated Producing Establishments, 2005 through 2017.

ABSTRACT: Ready-to-eat (RTE) meat and poultry product samples collected between 2005 and 2017 from RTE-producing establishments for the U.S. Department of Agriculture, Food Safety and Inspection Service (FSIS) ALLRTE/RTEPROD_RAND (random) and RTE001/RTEPROD_RISK (risk-based) sampling projects were tested for Listeria monocytogenes (Lm). Data for 45,897 ALLRTE/RTEPROD_RAND samples collected from 3,607 distinct establishments and 112,347 RTE001/RTEPROD_RISK samples collected from 3,283 distinct establishments were analyzed for the presence of Lm. These data were also analyzed based upon the percentages of establishments with positive samples, annual production volume, sanitation control alternatives, geographic location, and season or month of sample collection. Results revealed low occurrence of Lm-positive samples from the random and risk-based sampling projects, with 152 (0.33%) positive samples for ALLRTE/RTEPROD_RAND and 403 (0.36%) positive samples for RTE001/RTEPROD_RISK. The percentage of positive samples significantly decreased over time, from about 0.7% in 2005 and 2006 to about 0.2% in 2017 (P < 0.05). From 2005 to 2017, 3.9% of establishments sampled under the ALLRTE/RTEPROD_RAND sampling project had at least one Lm-positive sample. Similarly, 10.0% of establishments sampled under the RTE001/RTEPROD_RISK sampling project had at least one positive sample. Samples positive for Lm were found in all geographic regions in all months. Thus, in 13 years of RTE product sampling in FSIS-regulated establishments (2005 through 2017), <0.4% of samples were positive for Lm in both risk-based and random sampling projects. The low prevalence of Lm in these products suggests that the combination of FSIS policies and industry practices may be effective for controlling Lm contamination. Information obtained from these sampling projects is relevant to the ongoing prevention of foodborne Lm illnesses from RTE meat and poultry products.

Predictive Model for Growth of Bacillus cereus at Temperatures Applicable to Cooling of Cooked Pasta.

A model was developed to predict the growth of Bacillus cereus from spores during cooling of cooked pasta. Cooked pasta was inoculated with a cocktail of four strains of heat-shocked (80 °C/10 min) B. cereus spores to obtain a final spore concentration of approximately 2 log CFU/g. Thereafter, growth was determined at isothermal temperatures starting at 10 °C and every three degrees up to 49 °C. Samples were removed periodically and plated on mannitol egg yolk polymyxin agar. The plates were incubated for 24 hr at 30 °C. Baranyi, Huang, and modified Gompertz primary growth models were used to fit growth data. The modified Ratkowsky secondary model was used to fit growth rates determined by the primary growth models with respect to temperature. All three primary models fitted the growth data well. The modified Ratkowsky secondary model adequately fit growth rates generated by the three primary models (R2 values ranging from 0.96 to 0.98). After acceptable prediction zone (APZ) validation and goodness of fit statistical analyses, it was determined that the Baranyi primary growth model was best suited for these data. For both single-rate exponential cooling and biphasic linear cooling model validation, all Baranyi model predictions (n = 24 and 28, respectively) fell within the APZ (-1.0 to 0.5 log CFU/g). The model will assist institutional food service settings to determine the safety of cooked pasta subjected to longer cooling times or stored at improper temperatures. PRACTICAL APPLICATION: Predictive model can be used to estimate extent of microbial growth during cooling of cooked pasta and in designing HACCP program and setting of critical control levels. Retail food industry would need fewer challenge studies to validate the safety of their products. The model will provide regulatory agencies and food industry with an objective means of assessing the microbial risk and ensuring that the public is not at risk of acquiring food poisoning.

Predictive model for growth of Bacillus cereus during cooling of cooked rice.

Bacillus cereus is frequently implicated in foodborne outbreaks associated with the consumption of cooked rice. The main contributing factors leading to outbreaks is rice cooked in large quantities and subsequently, inadequately chilled or stored at room temperatures for a prolonged period of time prior to consumption. Bacillus cereus growth in cooked rice inoculated with approximately 2 log CFU/g of heat-shocked (80 °C/10 min) spores at several isothermal conditions (between 10 and 49 °C) was quantified. B. cereus populations were determined by plating on mannitol egg yolk polymyxin agar and incubating at 30 °C for 24 h. Primary growth models, namely Baranyi, Huang, modified Gompertz, and logistic models were fitted to growth data. Specific growth rates from all four primary models were used to fit the modified Ratkowsky square-root model with respect to temperature. All four primary models were well fitted by the modified Ratkowsky model (R2 values from 0.90-0.99). Based on the goodness of fit secondary model statistics (R2, SSE, RMSE), the Baranyi model performed the best and was chosen for tertiary modeling. Acceptable prediction zone (APZ) analysis was performed for validation of the Baranyi model predictions during single rate exponential and biphasic linear cooling temperature profiles. For single rate cooling, 23 of the 24 predictions fell within the APZ (-1.0 to 0.5 log CFU/g). For biphasic linear cooling, 26 of the 28 predictions fell within the APZ. The developed dynamic model can be used to predict potential B. cereus growth from spores in cooked rice during chilling and thus, support the disposition of product subject to cooling deviations.

Occurrence of Salmonella in Ready-to-Eat Meat and Poultry Product Samples from U.S. Department of Agriculture-Regulated Producing Establishments. I. Results from the ALLRTE and RTE001 Random and Risk-Based Sampling Projects, from 2005 to 2012.

Ready-to-eat (RTE) meat and poultry product samples collected from RTE-producing establishments for the ALLRTE (random) and RTE001 (risk-based) sampling projects of the Food Safety and Inspection Service (FSIS) were tested for both Salmonella and Listeria monocytogenes. The FSIS analyzed Salmonella results for RTE meat and poultry product samples collected for the two sampling projects from 2005 to 2012. Data for 24,385 ALLRTE samples collected from 3,023 establishments and 66,653 RTE001 samples collected from 2,784 establishments were evaluated for the percentages of Salmonella-positive samples, product types of positive samples, and Salmonella serotypes. There also were descriptive summaries with respect to establishment hazard analysis and critical control point (HACCP) size, production volumes, L. monocytogenes control alternatives, geographic location, and season or month of sample collection. Results showed low occurrences of Salmonella-positive samples from the ALLRTE and RTE001 sampling projects, with 14 positive samples (0.06%) for ALLRTE and 33 positive samples (0.05%) for RTE001. Percentages of establishments with at least one Salmonella-positive sample averaged 0.46% for ALLRTE and 1.11% for RTE001. Three product types-sausage products, pork barbecue, and head cheese-accounted for 62% of all positive samples. There were 27 distinct serotypes from 48 Salmonella isolates, with serotypes Infantis and Typhimurium being the most common (5 isolates each). All but one of the Salmonella-positive samples were obtained from establishments with HACCP sizes of small or very small. More than half of the positive samples were obtained from establishments using L. monocytogenes control alternative 3 (sanitation only, highest-risk category). Positive Salmonella samples were found in all geographic regions at all times of the year. Information obtained from these sampling projects is relevant to the prevention of foodborne Salmonella illnesses from RTE meat and poultry products.

Occurrence of Salmonella in Ready-to-Eat Meat and Poultry Product Samples from U.S. Department of Agriculture-Regulated Producing Establishments. II. Salmonella in Ready-to-Eat Pork Barbecue Products, from 2005 to 2012.

Ready-to-eat (RTE) meat and poultry product samples from the random ALLRTE and risk-based RTE001 sampling projects of the Food Safety and Inspection Service (FSIS) were tested for both Listeria monocytogenes and Salmonella. In the course of analyzing Salmonella data for calendar years 2005 to 2012, it was observed that 8 (17.0%) of 47 positive samples were from pork barbecue. The eight Salmonella-positive samples, from seven establishments in a single state, were from 1,085 pork barbecue samples tested nationwide (0.74% positive) and from 296 samples tested from that one state (2.7% positive). The seven establishments represented 30.4% of 23 federal establishments in that state that had pork barbecue samples tested for Salmonella. A follow-up sample from intensified verification testing at one of the seven establishments also was positive for Salmonella. Upon further examination, contamination appeared to be influenced by regional differences in production methods. Notably, the style of pork barbecue that tested positive for Salmonella used a vinegar- and pepper-based sauce in which the ingredients were mixed without cooking. All the establishments with Salmonella-positive samples followed the practice of first cooking the pork and then adding the barbecue sauce ingredients (vinegar, pepper, other spices, etc.) after cooking (postlethality exposure). In addition to the sauce ingredients, other possible sources of contamination included employee hygiene and food handling practices and cross-contamination from other Salmonella-contaminated products and from commonly used equipment. Based on these findings, the FSIS issued guidelines recommending changes in production methods that would minimize or eliminate pork barbecue as a potential source of foodborne Salmonella infections.

Influence of Cooling Rate on Growth of Bacillus cereus from Spore Inocula in Cooked Rice, Beans, Pasta, and Combination Products Containing Meat or Poultry.

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 <1 log after 9 h of cooling, but the pathogen grew faster in rice and pasta. In combination products, the net growth was as follows: 3.05, 3.89, and 4.91 log CFU/g in rice-chicken; 3.49, 4.28, and 4.96 log CFU/g in rice-beef; 3.50, 4.20, and 5.32 CFU/g in rice-chicken-mixed vegetables; and 3.68, 4.44, and 5.25 CFU/g in rice-beef-mixed vegetables after 15, 18, and 21 h of cooling, respectively. This study suggests safe cooling rates for cooling cooked rice, beans, pasta, rice-chicken, rice-chicken-vegetables, rice-beef, and rice-beef-vegetables to guard against the hazards associated with B. cereus.

Fate of Shiga toxin-producing O157:H7 and non-O157:H7 Escherichia coli cells within refrigerated, frozen, or frozen then thawed ground beef patties cooked on a commercial open-flame gas or a clamshell electric grill.

Both high-fat and low-fat ground beef (percent lean:fat = ca. 70:30 and 93:7, respectively) were inoculated with a 6-strain cocktail of non-O157:H7 Shiga toxin-producing Escherichia coli (STEC) or a five-strain cocktail of E. coli O157:H7 (ca. 7.0 log CFU/g). Patties were pressed (ca. 2.54 cm thick, ca. 300 g each) and then refrigerated (4°C, 18 to 24 h), or frozen (-18°C, 3 weeks), or frozen (-18°C, 3 weeks) and then thawed (4°C for 18 h or 21°C for 10 h) before being cooked on commercial gas or electric grills to internal temperatures of 60 to 76.6°C. For E. coli O157:H7, regardless of grill type or fat level, cooking refrigerated patties to 71.1 or 76.6°C decreased E. coli O157:H7 numbers from an initial level of ca. 7.0 log CFU/g to a final level of ≤1.0 log CFU/g, whereas decreases to ca. 1.1 to 3.1 log CFU/g were observed when refrigerated patties were cooked to 60.0 or 65.5°C. For patties that were frozen or freeze-thawed and cooked to 71.1 or 76.6°C, E. coli O157:H7 numbers decreased to ca. 1.7 or ≤0.7 log CFU/g. Likewise, pathogen numbers decreased to ca. 0.7 to 3.7 log CFU/g in patties that were frozen or freeze-thawed and cooked to 60.0 or 65.5°C. For STEC, regardless of grill type or fat level, cooking refrigerated patties to 71.1 or 76.6°C decreased pathogen numbers from ca. 7.0 to ≤0.7 log CFU/g, whereas decreases to ca. 0.7 to 3.6 log CFU/g were observed when refrigerated patties were cooked to 60.0 or 65.5°C. For patties that were frozen or freeze-thawed and cooked to 71.1 or 76.6°C, STEC numbers decreased to a final level of ca. 1.5 to ≤0.7 log CFU/g. Likewise, pathogen numbers decreased from ca. 7.0 to ca. 0.8 to 4.3 log CFU/g in patties that were frozen or freeze-thawed and cooked to 60.0 or 65.5°C. Thus, cooking ground beef patties that were refrigerated, frozen, or freeze-thawed to internal temperatures of 71.1 and 76.6°C was effective for eliminating ca. 5.1 to 7.0 log CFU of E. coli O157:H7 and STEC per g.

Growth potential of Clostridium perfringens from spores in acidified beef, pork, and poultry products during chilling.

The ability of Clostridium perfringens to germinate and grow in acidified ground beef as well as in 10 commercially prepared acidified beef, pork, and poultry products was assessed. The pH of ground beef was adjusted with organic vinegar to achieve various pH values between 5.0 and 5.6; the pH of the commercial products ranged from 4.74 to 6.35. Products were inoculated with a three-strain cocktail of C. perfringens spores to achieve ca. 2-log (low) or 4-log (high) inoculum levels, vacuum packaged, and cooled exponentially from 54.4 to 7.2°C for 6, 9, 12, 15, 18, or 21 h to simulate abusive cooling; the U.S. Department of Agriculture, Food Safety and Inspection Service (USDA-FSIS) recommends a cooling time of 6.5 h. Total germinated C. perfringens populations were determined after plating on tryptose-sulfite-cycloserine agar and incubating the plates anaerobically at 37°C for 48 h. In addition, C. perfringens growth from spores was assessed at an isothermal temperature of 44°C. Growth from spores was inhibited in ground beef with a pH of 5.5 or below, even during extended cooling from 54.4 to 7.2°C in 21 h. In ground beef with a pH of 5.6, the growth was >1 log after 18 h of cooling from 54.4 to 7.2°C. However, 15 h of cooling controlled the growth to <1 log, regardless of the inoculum level. In addition, no growth was observed in any product with a pH ranging from 4.74 to 5.17, both during exponential abusive cooling periods of up to 21 h and during storage for 21 h at 44°C. While <1-log growth of C. perfringens from spores was observed in the pH 5.63 product cooled exponentially from 54.4 to 7.2°C in 15 h or less, the pH 6.35 product supported growth, even after 6 h of cooling from 54.4 to 7.2°C. These challenge tests demonstrate that adjustment of ground beef to pH of 5.5 or less and of barbeque products to pH of 5.63 or less inhibits C. perfringens spore germination and outgrowth during extended cooling periods from 54.4 to 7.2°C up to 15 h. Therefore, safe cooling periods for products with homogeneous, lower pHs can be substantially longer.

Predictive thermal inactivation model for effects of temperature, sodium lactate, NaCl, and sodium pyrophosphate on Salmonella serotypes in ground beef.

Analyses of survival data of a mixture of Salmonella spp. at fixed temperatures between 55 degrees C (131 degrees F) and 71.1 degrees C (160 degrees F) in ground beef matrices containing concentrations of salt between 0 and 4.5%, concentrations of sodium pyrophosphate (SPP) between 0 and 0.5%, and concentrations of sodium lactate (NaL) between 0 and 4.5% indicated that heat resistance of Salmonella increases with increasing levels of SPP and salt, except that, for salt, for larger lethalities close to 6.5, the effect of salt was evident only at low temperatures (<64 degrees C). NaL did not seem to affect the heat resistance of Salmonella as much as the effects induced by the other variables studied. An omnibus model for predicting the lethality for given times and temperatures for ground beef matrices within the range studied was developed that reflects the convex survival curves that were observed. However, the standard errors of the predicted lethalities from this models are large, so consequently, a model, specific for predicting the times needed to obtained a lethality of 6.5 log(10), was developed, using estimated results of times derived from the individual survival curves. For the latter model, the coefficient of variation (CV) of predicted times range from about 6 to 25%. For example, at 60 degrees C, when increasing the concentration of salt from 0 to 4.5%, and assuming that the concentration of SPP is 0%, the time to reach a 6.5-log(10) relative reduction is predicted to increase from 20 min (CV = 11%) to 48 min (CV = 15%), a 2.4 factor (CV = 19%). At 71.1 degrees C (160 degrees F) the model predicts that more than 0.5 min is needed to achieve a 6.5-log(10) relative reduction.