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.

Effects of spore purity on the wet heat resistance of Clostridium perfringens, Bacillus cereus and Bacillus subtilis spores.

Heat resistance of spores of Clostridium perfringens 8238 (Hobbs Serotype 2), Bacillus cereus NCTC 11143 (4810/72), and Bacillus subtilis PS533, an isogenic derivative of strain PS832 (a 168 strain) was determined in ground beef at 95 °C. Spore purification was by centrifugation and washing with sterile distilled water (dH2O), followed by sonication and then Histodenz centrifugation for B. subtilis and C. perfringens, and centrifugation and washing with sterile dH2O followed by Histodenz centrifugation for B. cereus. Bags containing inoculated beef samples were submerged in a temperature-controlled water bath and held at 95 °C for predetermined lengths of time. Surviving spore populations were enumerated by plating on mannitol egg yolk polymyxin agar (MYP) agar plates for B. cereus and B. subtilis, and on tryptose-sulfite-cycloserine agar (TSC) agar plates for C. perfringens. Survivor curves were fitted to linear, linear with tail, and Weibull models using the USDA Integrated Pathogen Modeling Program (IPMP) 2013 software. The Weibull model provided a relatively better fit to the data since the root mean square error (RMSE), mean square error (MSE), sum of squared errors (SSE), and Akaike information criterion (AIC) values were lower than the values obtained using the linear or the linear with tail models. Additionally, the Weibull model accurately predicted the observed D-values at 95 °C for the three spore-formers since the accuracy factor (Af) values ranged from 1.03 to 1.08 and the bias factor (Bf) values were either 1.00 or 1.01. Times at 95 °C to achieve a 3-log reduction decreased from 206 min for C. perfringens spores purified with water washes alone to 191 min with water washes followed by sonication and Histodenz centrifugation, from 7.9 min for B. cereus spores purified with water washes alone to 1.4 min with water washes followed by Histodenz centrifugation, and from 20.6 min for B. subtilis spores purified with water washes alone to 6.7 min for water washes followed by sonication and Histodenz centrifugation. Thermal-death-time values reported in this study will assist food processors to design thermal processes to guard against bacterial spores in cooked foods. In addition, clearly spore purity is an additional factor in spore wet heat resistance, although the cause of this effect is not clear.

Optimizing the Effects of Nisin and NaCl to Thermal Inactivate Listeria monocytogenes in Ground Beef with Chipotle Sauce During Sous-vide Processing.

Mild cooking thermal treatments, like sous-vide, can compromise ground meat entrees such as meatballs with chipotle sauce, especially when salt levels are reduced during its preparation. Listeria monocytogenes is a thermoresistant pathogen that can be in ready-to-eat food. On the other hand, nisin, due to its thermal stability, can be a good alternative to aid on the thermal inactivation of L. monocytogenes and ensure meat safety. The objective was to optimize the amount of nisin and salt concentrations to thermally inactivate L. monocytogenes during the sous-vide cooking of ground beef marinated in chipotle sauce, and to generate a predictive model. A four-strain cocktail was prepared and inoculated in ground beef in combination (3:2) with chipotle sauce added with nisin (0-150 IU) and salt (0-2%). After that, meat samples were sous-vide cooked at different temperatures, nisin, and salt concentrations, established by a central composite design. Depending on the levels of these factors, D-values ranged from 49.71 to 0.27 min. A predictive model (p < 0.05) was obtained by response surface, which described that D-values variation was explained by the linear effects of the three factors, the interaction between nisin and temperature, and the quadratic effects of salt and temperature. It was also observed that nisin presented a bactericidal effect while salt presented a protective effect during the thermal inactivation of L. monocytogenes. Adding 120 IU of nisin and 0.4% of salt to the meat product at 63°C temperature can help to ensure food safety by making L. monocytogenes cells more sensitive to the lethal effect of heat. The model developed in this study can be used by food processors for planning and designing effective levels of salt and nisin to thermally inactivate L. monocytogenes in ground beef products marinated with chipotle sauce to ensure their safety.

Effect of Citral on the Thermal Inactivation of Escherichia coli O104:H4 in Ground Beef.

ABSTRACT: 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 (P < 0.05). A citral concentration-dependent effect (P < 0.05) 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.

Predictive model for growth of Clostridium botulinum from spores during cooling of cooked ground chicken.

Cooking temperature of poultry meat is typically inadequate to inactivate the heat resistant spores of Clostridium botulinum. The purpose of this study is to develop a predictive model for C. botulinum during cooling of cooked ground chicken. Cooked chicken was inoculated with a cocktail of five strains of proteolytic C. botulinum type A and five strains of proteolytic C. botulinum type B to yield a final spore concentration of approximately 2 log CFU/g. The growth of C. botulinum was determined at constant temperatures from 10 to 46 °C. Dynamic temperature experiments were performed with continued cooling from 54.4 to 4.4 °C or 7.2 °C in mono- or bi-phasic cooling profiles, respectively. The Baranyi primary model was used to fit growth data and the modified Ratkowsky secondary model was used to fit growth rates with respect to temperature. The primary models fitted the growth data well (R2 values ranging from 0.811 to 0.988). The R2 and root mean square error (RMSE) of the modified Ratkowsky secondary model were 0.95 and 0.06, respectively. Out of 11 prediction error values calculated in this study, ten were within the limit of acceptable prediction zone (-1.0 to 0.5), indicating a good fit of the model. The predictive model will assist institutional food service operations in determining the safety of cooked ground chicken subjected to different cooling periods.

Predictive model for growth of Clostridium perfringens during cooling of cooked pork supplemented with sodium chloride and sodium pyrophosphate.

A dynamic model was developed to predict growth of Clostridium perfringens in cooked ground pork supplemented with salt (0-3% wt/wt) and sodium pyrophosphate (0-0.3% wt/wt) under varying temperatures. C. perfringens (NCTC 8238, NCTC 8239, and NCTC 10240) spores were heat shocked, cooled, and inoculated into ground pork. Isothermal bacterial growth was quantified with variable salt and phosphate concentrations at temperatures ranging from 15 to 51 °C. The primary Baranyi model was fitted to all C. perfringens growth profiles and gave a satisfactory fit (R2 ≥ 0.85). A quadratic polynomial secondary model was developed (P < 0.0001) to predict the maximum specific growth rate as a function of temperature, salt, and phosphate concentrations (R2 = 0.93). A dynamic model was developed and validated using growth data retrieved from 7 published studies. Thirty three out of 44 predictions were within the acceptable prediction zone (-0.5 ≤ prediction error ≤ 1.0). The developed predictive model can be used to minimize the risk of C. perfringens in pork products supplemented with additives during cooling.

The effect of lauric arginate on the thermal inactivation of starved Listeria monocytogenes in sous-vide cooked ground beef.

The aim of this study was to examine the efficacy of lauric arginate (LAE, 1000 ppm - 3000 ppm) as an assisting tool to reduce starved Listeria monocytogenes population in ground beef following sous-vide processing at different temperatures (55-62.5 °C). Ground beef mixed with LAE was vacuum sealed and a laboratory water bath was used for sous-vide cooking. Loglinear and Weibull models were fit to the survival microbial population and the D and Z-values were determined at 55-62.5 °C. Calculated D-values ranged from 33.62 to 3.22 min at temperature 55-62.5 °C. LAE at higher concentration is an effective antimicrobial to increase the inactivation of the pathogen in sous-vide cooking. With the addition of LAE, D-values at 55 and 62.5 °C determined by the Loglinear model decreased from 31.86 to 2.28 min (LAE 1000 ppm) and 16.71 to 0.56 min (LAE 3000 ppm), respectively; whereas the D-values at 55 to 62.5 °C determined by the Weibull model were 44.26 and 2.09 min (LAE 1000 ppm) and 22.71 and 1.60 min (LAE 3000 ppm), respectively. This study shows that sous-vide processing of ground beef supplemented with higher concentration of LAE effectively inactivates L. monocytogenes and thus, helps increase the microbiological safety and product quality.

Effect of grape seed extract on heat resistance of Clostridium perfringens vegetative cells in sous vide processed ground beef.

The heat resistance (57.5-65 °C) of a three-strain cocktail of Clostridium perfringens vegetative cells in sous vide processed ground beef supplemented with 0-3% grape seed extract (GSE) was quantified. The surviving cell population was enumerated on tryptose-sulfite-cycloserine agar. The decimal reduction (D)-values in beef that included no GSE were 67.11, 17.15, 4.02, and 1.62 min at 57.5, 60, 62.5, and 65 °C, respectively. Addition of 1.0% GSE resulted in concomitant decrease in heat resistance as evidenced by reduced bacterial D-values. The D-values in beef with added 1.0% GSE were 62.89, 13.70, 3.47 and 1.46 min at 57.5, 60, 62.5, and 65 °C, respectively. The heat resistance was further decreased when the GSE concentration in beef was increased to 2 or 3%. The z-values in beef with or without GSE were similar, ranging from 4.41 to 4.56 °C. The results of this study would be beneficial to the retail and institutional food service establishments in estimating re-heating time and temperature for sous vide processed ground beef to ensure safety against C. perfringens.

Cross-Laboratory Comparative Study of the Impact of Experimental and Regression Methodologies on Salmonella Thermal Inactivation Parameters in Ground Beef.

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.