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.

One-step analysis of growth kinetics of mesophilic Bacillus cereus in liquid egg yolk during treatment with phospholipase A2: Model development and validation.

Liquid egg yolk (LEY) is often treated with phospholipase A2 (PLA2) to improve its emulsifying capacity and thermal stability. However, this process may allow certain pathogens to grow. The objective of this study was to evaluate the growth kinetics of mesophilic Bacillus cereus in LEY during PLA2 treatment. Samples, inoculated with B. cereus vegetative cells, were incubated isothermally at different temperatures between 9 and 50 °C to observe the bacterial growth and survival. Under the observation conditions, bacterial growth occurred between 15 and 48 °C, but not at 9 and 50 °C. The growth curves were analyzed using the USDA IPMP-Global Fit, with the no-lag phase model as the primary model in combination with either the cardinal temperatures model (CTM) or the Huang square-root model (HSRM) as the secondary model. While similar maximum growth temperatures (Tmax) were determined (48.4 °C for HSRM and 48.1 °C for CTM), the minimum growth temperature (Tmin) of the HSRM more accurately described the lower limit (9.26 °C), in contrast to 6.51 °C for CTM, suggesting that the combination of the no-lag phase model and HSRM was more suitable to describe the growth of mesophilic B. cereus in LEY. The root mean square error (RMSE) of model validation and development was <0.5 log CFU/g, indicating the combination of the no-lag phase model and HSRM could predict the growth of mesophilic B. cereus in LEY during PLA2 treatment. The results of this study may allow the food industry to choose a suitable temperature for PLA2 treatment of LEY to prevent the growth of mesophilic B. cereus.

Predictive modeling of thermal inactivation of non-O157 Shiga toxin-producing Escherichia coli in ground beef with varying fat contents.

A mathematical model to predict the thermal inactivation of non-O157 Shiga toxin-producing Escherichia coli (STEC) in ground beef was developed, with temperature and fat content of ground beef as controlling factors. Survival curves for a cocktail of non-O157 STEC strains in ground beef at four temperatures (55, 60, 65, and 68 °C) and six fat levels (5, 10, 15, 20, 25, and 30%) were generated. Nine primary models-log-linear, log-linear with tail, biphasic, sigmoidal, four-factor sigmoidal, Baranyi, Weibull, mixed Weibull, and Gompertz-were tested for fitting the survival curves. Primary modeling analysis showed the Weibull model had the highest accuracy factor and Akaike's weight, making it the best-fitting model. The parameters of the Weibull model were estimated using a nonlinear mixed, and response surface modeling was used to develop a second-order polynomial regression to estimate the impact of fat in ground beef and cooking temperature on the heat resistance of non-O157 STEC strains. The secondary model was successfully validated by comparing predicted lethality (log10 CFU/g) with the observed values for ground beef containing 10 and 27% fat at 58 and 62 °C. Process lethality obtained from experimental data was within the prediction interval of the predictive model. The developed model will assist the food industry in estimating the appropriate time and temperature required for cooking ground beef to provide adequate protection against STEC contaminants.

Molecular modeling, simulation and docking of Rv1250 protein from Mycobacterium tuberculosis.

Computational prediction and protein structure modeling have come to the aid of various biological problems in determining the structure of proteins. These technologies have revolutionized the biological world of research, allowing scientists and researchers to gain insights into their biological questions and design experimental research much more efficiently. Pathogenic Mycobacterium spp. is known to stay alive within the macrophages of its host. Mycobacterium tuberculosis is an acid-fast bacterium that is the most common cause of tuberculosis and is considered to be the main cause of resistance of tuberculosis as a leading health issue. The genome of Mycobacterium tuberculosis contains more than 4,000 genes, of which the majority are of unknown function. An attempt has been made to computationally model and dock one of its proteins, Rv1250 (MTV006.22), which is considered as an apparent drug-transporter, integral membrane protein, and member of major facilitator superfamily (MFS). The most widely used techniques, i.e., homology modeling, molecular docking, and molecular dynamics (MD) simulation in the field of structural bioinformatics, have been used in the present work to study the behavior of Rv1250 protein from M. tuberculosis. The structure of unknown TB protein, i.e., Rv1250 was retrived using homology modeling with the help of I-TASSER server. Further, one of the sites responsible for infection was identified and docking was done by using the specific Isoniazid ligand which is an inhibitor of this protein. Finally, the stability of protein model and analysis of stable and static interaction between protein and ligand molecular dynamic simulation was performed at 100 ns The designing of novel Rv1250 enzyme inhibitors is likely achievable with the use of proposed predicted model, which could be helpful in preventing the pathogenesis caused by M. tuberculosis. Finally, the MD simulation was done to evaluate the stability of the ligand for the specific protein.

Effect of Disulfide Bonds on the Thermal Stability of Pediocin: In-silico Screening Using Molecular Dynamics Simulation.

The thermal stability properties of pediocin at 310, 313, 323, 333, 343, and 348 K (37, 40, 50, 60, 70, and 75°C, respectively) are reported in this study. A theoretical approach, such as the molecular dynamics method, was used to analyze the structure. Molecular dynamics simulation confirms the stability of molecules with Cys. Furthermore, this study reveals that Cys residues play an essential role in structure stability at high temperatures. To understand the structural basis for the stability of pediocin, a detailed in-silico analysis using molecular dynamics simulations to explore the thermal stability profiles of the compounds was conducted. This study shows that thermal effects fundamentally alter the functionally crucial secondary structure of pediocin. However, as previously reported, pediocin's activity was strictly conserved due to the disulfide bond between Cys residues. These findings reveal, for the first time, the dominant factor behind the thermodynamic stability of pediocin.

Modeling the Fate of Listeria monocytogenes and Salmonella enterica on Fresh Whole and Chopped Wood Ear and Enoki Mushrooms.

Two recent foodborne illness outbreaks linked to specialty mushrooms have occurred in the United States, both representing novel pathogen-commodity pairings. Listeria monocytogenes and Salmonella enterica were linked to enoki and wood ear mushrooms, respectively. The aim of this study was therefore to examine the survival of both L. monocytogenes and S. enterica on raw whole and chopped enoki and wood ear mushrooms during storage at different temperatures. Fresh mushrooms were either left whole or chopped and subsequently inoculated with a cocktail of either S. enterica or rifampicin-resistant L. monocytogenes, resulting in an initial inoculation level of 3 log CFU/g. Mushroom samples were stored at 5, 10, or 25°C for up to 7 d. During storage, the population levels of S. enterica or L. monocytogenes on the mushrooms were enumerated. The primary Baranyi model was used to estimate the growth rates of both pathogens and the secondary Ratkowsky square root model was used to model the relationship between growth rates and temperature. Both L. monocytogenes and S. enterica survived on both mushroom types and preparations at all temperatures. No proliferation of either pathogen was observed on mushrooms stored at 5°C. At 10°C, moderate growth was observed for both pathogens on enoki mushrooms and for L. monocytogenes on wood ear mushrooms; no growth was observed for S. enterica on wood ear mushrooms. At 25°C, both pathogens proliferated on both mushroom types with growth rates ranging from 0.43 to 3.27 log CFU/g/d, resulting in 1 log CFU/g increases in only 0.31 d (7.44 h) to 2.32 d. Secondary models were generated for L. monocytogenes on whole wood ear mushrooms and S. enterica on whole enoki mushrooms with goodness-of-fit parameters of r2 = 0.9855/RMSE = 0.0479 and r2 = 0.9882/RMSE = 0.1417, respectively. Results from this study can aid in understanding the dynamics of L. monocytogenes and S. enterica on two types of specialty mushrooms.

Ready-to-Eat Egg Products Formulated with Nisin and Organic Acids to Control Listeria monocytogenes.

Formulating ready-to-eat (RTE) products with growth inhibitors minimizes the risk of listeriosis. In part I, RTE egg products formulated with 6.25 ppm nisin were evaluated to control Listeria monocytogenes. Individual experimental units were surface inoculated with 2.5-log CFU/g of L. monocytogenes, packaged in pouches with a headspace gas of 20:80 CO2:NO2, and stored at 4.4°C for 8 weeks. Formulations with finished product pH of 6.29 ± 0.07 limited growth to <2-log for 4 weeks. Products at pH values of 7.42 ± 0.12 and 7.84 ± 0.11 were not different (p > 0.05) from the control without nisin at pH 7.34 ± 0.13, all supported 4-log growth by 4 weeks. In part II, a nisin bioassay test was performed to evaluate the stability of nisin in eggs as affected by the product pH (6.00 ± 0.03, 7.00 ± 0.00, 7.50 ± 0.03, and 8.00 ± 0.02) and cooking to an internal temperature of 73.9 or 85°C for 90 s. The nisin activity loss increased as the product pH or the cooking temperature increased (p < 0.05). Part III evaluated the effectiveness of 6.25 ppm nisin in combination with either an acetate-based antimicrobial used at 1.0% (w/w) in egg formulation (A1.0), propionate at 0.3% (P0.3), acetate-diacetate at 1.0% (AD1.0), acetate-diacetate at 0.6% (AD0.6), and lactate at 2.0% (L2.0) as a positive control. These formulations had a finished product pH, moisture, and salt contents of 5.97 ± 0.21, 72.4 ± 0.9%, and 0.67 ± 0.05%, respectively. L. monocytogenes did not grow in formulations A1.0 and AD1.0, whereas L2.0 and P0.3 supported 2-log growth by weeks 6 and 15, respectively and AD0.6 supported <1-log growth over 20 weeks at 4.4°C. Evaluation of uninoculated control units in parts I and III showed no changes (p > 0.05) in the CO2 and O2 headspace gas composition, generally no detection or growth of background microbes, and no changes (p > 0.05) in the pH of the formulations during storage, all assuring absence of uncontrolled interferences for the growth of L. monocytogenes.

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.

Antibiotic Resistance Influences Growth Rates and Cross-Tolerance to Lactic Acid in Escherichia coli O157:H7 H1730.

Escherichia coli O157:H7-contaminated beef has been implicated in numerous foodborne outbreaks. Contamination occurs despite the use of antimicrobial interventions such as lactic acid (LA). In addition, resistance to antibiotics such as ampicillin and streptomycin among isolates has been frequently reported. The influence of antibiotic resistance (ABR) on growth rates and cross-tolerance of lettuce isolate E. coli O157:H7 H1730 to LA was evaluated. Antibiotic-resistant strain variants were generated by conferring resistance to either ampicillin (ampC) or streptomycin (strepC) or both ampicillin and streptomycin (ampC strepC) through incremental exposure to the antibiotics. Ampicillin resistance was also conferred by plasmid transformation to generate the ampP and ampP strepC strains. The minimum inhibitory concentration of LA on all the strains evaluated was 0.375% v/v. The lag phase duration of all strains except E. coli O157:H7 ampP strepC increased with increasing concentration of LA. The ampP strepC and ampC strains were most tolerant to 5% LA with declines in the cell population of 2.86 and 2.56 log CFU/mL, respectively (p < 0.05). The ampP strepC strain was the most tolerant when evaluated by the live/dead viability assay. The addition of the efflux pump inhibitor, carbonyl cyanide m-chlorophenylhydrazone, with 2.5% LA resulted in a significant increase in sensitivity in the no resistance (NR) wild-type and ampC strains, resulting in 6.62 and 6.65 log CFU/mL reduction, respectively, while the highly tolerant ampP strepC strain had a 2.90 log CFU/mL decrease. Tolerance to LA was significantly influenced by both the ABR profile of the strain and LA concentration. The results from this study indicate that E. coli O157:H7 strains with certain ABR profiles might be more tolerant to LA.

Advances in multi-omics based quantitative microbial risk assessment in the dairy sector: A semi-systematic review.

With the increasing consumption of packaged and ready-to-eat food products, the risk of foodborne illness has drastically increased and so has the dire need for proper management. The conventional Microbial Risk Assessment (MRA) investigations require prior knowledge of process flow, exposure, and hazard assessment throughout the supply chain. These data are often generated using conventional microbiological approaches based either on shelf-life studies or specific spoilage organisms (SSOs), frequently overlooking crucial information such as antimicrobial resistance (AMR), biofilm formation, virulence factors and other physiological variations coupled with bio-chemical characteristics of food matrix. Additionally, the microbial risks in food are diverse and heterogenous, that might be an outcome of growth and activity of multiple microbial populations rather than a single species contamination. The uncertainty on the microbial source, time as well as point of entry into the food supply chain poses a constraint to the efficiency of preventive approaches and conventional MRA. In the last few decades, significant breakthroughs in molecular methods and continuously progressing bioinformatics tools have opened up a new horizon for risk analysis-based approaches in food safety. Real time polymerase chain reaction (qPCR) and kit-based assays provide better accuracy and precision with shorter processing time. Despite these improvements, the effect of complex food matrix on growth environment and recovery of pathogen is a persistent problem for risk assessors. The dairy industry is highly impacted by spoilage and pathogenic microorganisms. Therefore, this review discusses the evolution and recent advances in MRAmethodologies equipped with predictive interventions and "multi-omics" approach for robust MRA specifically targeting dairy products. It also highlights the limiting gap area and the opportunity for improvement in this field to ensure precision food safety.

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.

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.

Inactivation of Salmonella and Shiga toxin-producing Escherichia coli (STEC) from the surface of alfalfa seeds and sprouts by combined antimicrobial treatments using ozone and electrolyzed water.

Individual chemical and non-chemical treatments have failed to disinfect alfalfa seeds and sprouts from pathogens thoroughly. This study investigated the disinfection of alfalfa seeds and sprouts using a procedure combining ozone with acidic (pH 3.0) electrolyzed water (AEW). Inoculated alfalfa seeds with a cocktail of 3 strains Salmonella and 3 strains of STEC were treated sequentially with aqueous ozone followed by AEW. Treatment started by immersing the samples into ozonated water (5 mg/L ozone) for 15 or 20 min with persistent oxygen feeding pressurized with 10 psi. The samples then were immersed in 1 L of AEW for 15 min. Salmonella and STEC were significantly (P < 0.05) reduced by 3.6 and 2.9 log CFU/g on seeds respectively, and by 3.1 and 3.0 log CFU/g reduction on sprouts. Significant differences (P < 0.05) were found in the magnitude of the log reduction between Salmonella and STEC on seeds and between seeds and sprouts. Using combined treatments showed no significant changes in the quality, including shelf life, weight, and color in sprouts as compared to controls. The findings suggest that the combination of ozone and AEW is effective in inactivation of Salmonella and STEC on alfalfa seeds and sprouts with no adverse effects on sprouts quality.

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.

Development of sodium chlorite and glucono delta-lactone incorporated PLA film for microbial inactivation on fresh tomato.

Chlorine dioxide (ClO2) is an effective disinfectant used in the sanitization of fresh produce. Glucono delta-lactone (GDL), widely used as an acidifier during food processing, can be partially hydrolyzed to become a weak acid-gluconic acid under chemical equilibrium upon dissolution in water. This study focused on the development of a novel polylactic acid (PLA) film which incorporated with sodium chlorite (NaClO2) and GDL for ClO2(g) generation. The effects of PLA amount, NaClO2 + GDL/PLA ratio, NaClO2/GDL ratio, temperature and relative humidity on the release profiles of ClO2(g) were elucidated. The storage test indicated that film efficacy was well maintained after 4 weeks of storage under ambient conditions. The microbial inactivation results revealed that ClO2(g) generated from the films reduced populations of surface-inoculated Salmonella and Escherichia coli O157:H7 from ca. 5 log CFU/tomato to undetectable level (<1 log CFU/tomato) within 2 and 4 h respectively and the complete elimination in populations of both bacterial species was maintained throughout the 14-day storage period at both 10 and 22 °C. The sensory properties of treated tomatoes were evaluated and exhibited no significant difference (p > 0.05) compared to controls except for appearance on day 14 under 22 °C storage.

Thermal Inactivation Kinetics of Three Heat-Resistant Salmonella Strains in Whole Liquid Egg.

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.

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.