Thermal resistance of Listeria monocytogenes, Escherichia coli O157:H7 and Salmonella in animal fat - Kinetic analysis and mathematical modeling.

The objective of this study was to evaluate the effect of fat on thermal resistance of L. monocytogenes, E. coli O157:H7, and Salmonella spp. A 4-strain cocktail of each microorganism was inoculated to beef tallow and heated isothermally at temperatures between 55 and 80℃. All survival curves did not follow the 1st-order inactivation kinetics but conformed to a two-stage linear pattern. The first stage was markedly less heat-resistant than the second, as manifested by significantly lower D values. The z values of E. coli O157 H7 and Salmonella spp. were 11.8 °C and 12.3 °C in the first stage (z1) but increased to 23.7 °C and 20.8 °C in the second stage (z2), respectively. For L. monocytogenes, while the z values were similar for both stages (z1 = 19.6 °C and z2 = 18.5 °C), the second stage D values are 3.6-5.9 times of those in the first stage. One-step analysis was used to fit the nonlinear curves to the Weibull model, yielding < 1 exponents for the model (0.495, 0.362, and 0.282, respectively, for L. monocytogenes, E. coli O157:H7, and Salmonella spp.), suggesting gradually increased thermal resistance during heating. The experimental results showed that these microorganisms could resist heating for longer time and at higher temperatures in tallow than they do in regular meats containing lower levels of fat. The kinetic models can be used to develop thermal processes to properly inactivate pathogens contaminated in the fat portions of meat products or other high fat products.

Numerical simulation of heat transfer during meat ball cooking and microbial food safety enhancement.

This study was conducted to apply the finite volume method (FVM) to solve the partial differential equation (PDE) governing the heat transfer process during meat cooking with convective surface conditions. For a one-dimensional, round-shaped food, such as meat balls, the domain may be divided into shells of equal thickness, with energy balance established for each adjacent shell using in the finite difference scheme (FDS) to construct a set of finite difference equations, which were then solved simultaneously using the FORTRAN language and the IVPAG subroutine of the International Mathematics and Statistics Library. The FDS is flexible for temperature-dependent physical properties of foods, such as thermal conductivity (k), specific heat (Cp ), thermal diffusivity (α), and boundary conditions, for example, surface heat transfer coefficient (h), to predict the dynamic temperature profiles in beef and chicken meat balls cooked in an oven. Once the FVM model was established and validated, it was used to simulate the dynamic temperature profiles during cooking, which were then used in combination with the general method to evaluate the thermal lethality of Shiga toxin-producing Escherichia coli and Salmonella spp. using D and z values in ground meats during cooking. The method can be applied to design cooking processes that effectively inactivate foodborne pathogens while maintaining the quality of cooked meats and evaluate the adequacy of a cooking process. PRACTICAL APPLICATION: The temperature dependences of thermal conductivity (k) and thermal diffusivity (α) of raw ground beef and ground chicken meats were measured. These thermal properties were then used in numerical simulation to predict the dynamic heating temperature profile and thermal lethality of ground beef and chicken meat balls. The numerical simulation method may be used to optimize and evaluate thermal processes and ensure the inactivation of pathogens in meat products during cooking.

Shelf-life boundaries of Listeria monocytogenes in cold smoked salmon during refrigerated storage and temperature abuse.

Cold smoked salmon (CSS) is a high-value ready-to-eat product, but it generally has a short shelf-life even under refrigeration and can support the growth of Listeria monocytogenes. Therefore, the objective of this study was to examine the growth and survival of L. monocytogenes in CSS during refrigerated storage and temperature abuse. The growth and survival data of L. monocytogenes (116 records, 465 data points) were retrieved from ComBase (https://www.combase.cc). All records contained storage time and temperature, but other information (aw, pH, and salt) was not fully documented. Each data point, normalized with the initial population to calculate relative growth (RG, log CFU/g), was used to classify the probability of growth. Eighty percent (80%) of the data were randomly sampled for examining the effect of storage time and temperature on growth of L. monocytogenes, while the remaining 20% were set aside for model validation. Logistic regression was used to develop a model for classifying L. monocytogenes growth according to 7 different control thresholds (CT), ranging from 0 to 3 log CFU/g in RG. A probability threshold was set to judge if the bacterial growth has exceeded a CT. The validation showed > 89% of true negative rate for not exceeding the control thresholds. A dynamic method was then developed and demonstrated to predict the growth probabilities under fluctuating temperature conditions. The result of this study suggested that storage time and temperature could be used to predict the growth of L. monocytogenes in CSS and to control listeriosis using a risk-based strategy. It can be used by the retailers and consumers to determine if a packaged product is safe to consume based on its time and temperature history.

Draft Genomic Sequence for Klebsiella pneumoniae 060517CS3-g, a Bacterium Harboring Multidrug Resistance Genes, Isolated from Retail Ground Chicken Meat.

Klebsiella pneumoniae is an important foodborne pathogen that can cause human infections. Here, we report the draft genomic sequence for K. pneumoniae 060517CS3-g, isolated from retail ground chicken meat, which has several antibiotic resistance genes, multiple plasmids, and genes that may result in its hypervirulence based on the sequence data.

Synergistic effect of high hydrostatic pressure, allyl isothiocyanate, and acetic acid on the inactivation and survival of pathogenic Escherichia coli in ground chicken.

Meat and poultry are prone to contamination with foodborne pathogens sourced from the livestock or introduced from the processing environments. In this study, for retention of meat quality while assuring microbial food safety, mild levels of high hydrostatic pressure were hurdled with food-grade additives (i.e., allyl isothiocyanate [AITC] and acetic acid [AA], functioned as antimicrobials) to inactivate pathogenic Escherichia coli in ground chicken. The reductions of Shiga toxin-producing E. coli (STEC) O157:H7 and uropathogenic E. coli (UPEC) were described as a function of high hydrostatic pressure (200-350 MPa), process-holding time (10-25 min), AITC concentration (0.05-0.20% w/w), and AA concentration (0.10--0.30% w/w) using a full factorial design. The antimicrobials had little influence on bacterial inactivation without high pressure. Without the antimicrobials, a high-pressure treatment at 300 MPa and 4°C for 15 min reduced E. coli O157:H7 and UPEC by 1.52 and 2.52 log, respectively. A 5-log reduction was achieved when AITC and AA were combined with high pressure, indicating a synergistic effect. The survivors were further reduced to below the detection limit of 1 log CFU/g after subsequent storage tests at 4 and 10°C for 10 days. The STEC O157:H7 was found slightly more resistant than UPEC in our test matrix. The developed models showed good fits with experimental data (R2  > 0.95 for linear models; Pr > F (<0.0001) for dimensionless nonlinear models); which may help processors find/optimize the processing parameters to achieve target foodborne pathogens reduction for food safety requirement. PRACTICAL APPLICATION: Models were developed to predict the inactivation of pathogenic Escherichia coli in ground chicken by high-pressure processing (HPP) in combination with natural antimicrobial compounds. These models can be used to estimate/determine the HPP operation parameters and antimicrobial usage levels (i.e., allyl isothiocyanate and acetic acid) needed to achieve a specific microbial log reduction within the selected factor ranges. The operation parameters and clean-label ingredients are of interest in the food industry, which may benefit from the application of the models in achieving microbial safety, process optimization, and operation cost reduction.

Modeling the effect of simultaneous use of allyl isothiocyanate and cinnamaldehyde on high hydrostatic pressure inactivation of Uropathogenic and Shiga toxin-producing Escherichia coli in ground chicken.

BACKGROUND: A combination of high-pressure processing (HPP) and antimicrobials is a well-known approach for enhancing the microbiological safety of foods. However, few studies have applied multiple antimicrobials simultaneously with HPP, which could be an additional hurdle for microbial inactivation. The present study applied a full factorial design to investigate the impact of HPP (225-325 MPa; 10-20 min), allyl isothiocyanate (AITC) (0.3-0.9 g kg-1 ) and trans-cinnamaldehyde (tCinn) (1.0-2.0 g kg-1 ) on the inactivation of Shiga toxin-producing Escherichia coli (STEC) O157:H7 and uropathogenic E. coli (UPEC) in ground chicken meat. RESULTS: The regulatory requirement of 5-log reduction was achieved at 305 MPa, 18 min, 0.8 g kg-1 AITC and 1.7 g kg-1 tCinn for STEC O157:H7 and at 293 MPa, 16 min, 0.6 g kg-1 AITC and 1.6 g kg-1 tCinn for UPEC, as specified by response surface analysis and verified via experiments. The surviving population was eliminated by post-treatment storage of 9 days at 10 °C. The developed linear regression models showed r2 > 0.9 for the E. coli inactivation. The developed dimensionless non-linear regression models covered a factorial range slightly wider than the original experimental limit, with probability Pr > F (< 0.0001). CONCLUSION: Simultaneous use of AITC and tCinn reduced not only the necessary concentration of each compound, but also the intensity of high-pressure treatments, at the same time achieving a similar level of microbial inactivation. STEC O157:H7 was found to be more resistant than UPEC to the HPP-AITC-tCinn stress. The developed models may be applied in commercial application to enhance the microbiological safety of ground chicken meat. Published 2020. This article is a U.S. Government work and is in the public domain in the USA.

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.

Effect of temperature on the growth of Staphylococcus aureus in ready-to-eat cooked rice with pork floss.

Cooked rice with pork floss (CRPF) wrapped in dried seaweed is one of the most popular ready-to-eat (RTE) foods in many Asian countries, particularly in Taiwan. The products are susceptible to Staphylococcus aureus contamination and temperature abuse during manufacturing, distribution, and storage. The objective of this study was to examine the effect of temperature on its growth in RTE CRPF for use in risk assessment and prevention of staphylococcal food poisoning (SFP). Inoculated CRPF samples were stored at 4, 12, 18, 25, and 35°C, and the change in the populations of S. aureus during storage were analyzed using three primary models to determine specific growth rate (µmax), lag-phase duration (λ), and maximum population density (ymax). The Ratkowsky square-root and Huang square-root (HSR) models were used as the secondary models to describe the effect of temperature on µmax, and a linear and an exponential regression models were used to describe the effect of temperature on λ and ymax, respectively. The model performance was evaluated by the root mean square error (RMSE), bias factor (Bf), and accuracy factor (Af) when appropriate. Results showed that three primary models were suitable for describing the growth curves, with RMSE ≤ 0.3 (log MPN/g). Using µmax obtained from the Huang model, the minimum growth temperature (Tmin) estimated by the HSR model was 7.0°C, well in agreement with the reported Tmin. The combination of primary and secondary models for predicting S. aureus growth was validated by additional growth curves at 30°C, which showed that the RMSE was 0.6 (log MPN/g). Therefore, the developed models were acceptable for predicting the growth of S. aureus in CRPF under likely temperature abuse conditions and can be applied to assess the risk of S. aureus in CRPF and design temperature controls to reduce the risk of SFP.

Survival Evaluation of Salmonella and Listeria monocytogenes on Selective and Nonselective Media in Ground Chicken Meat Subjected to High Hydrostatic Pressure and Carvacrol.

High pressure processing (HPP) and treatment with the essential oil extract carvacrol had synergistic inactivation effects on Salmonella and Listeria monocytogenes in fresh ground chicken meat. Seven days after HPP treatment at 350 MPa for 10 min, Salmonella treated with 0.75% carvacrol was reduced to below the detection limit (1 log CFU/g) at 4°C and was reduced by ca. 6 log CFU at 10°C. L. monocytogenes was more sensitive to these imposed stressors, remaining below the detection limit during storage at both 4 and 10°C after HPP treatment at 350 MPa for 10 min following treatment with 0.45% carvacrol. However, pressure-injured bacterial cells may recover and lead to an overestimation of process lethality when a selective medium is used without proper justification. For HPP-stressed Salmonella, a 1- to 2-log difference was found between viable counts on xylose lysine Tergitol 4 agar and aerobic plate counts, but no significant difference was found for HPP-stressed L. monocytogenes between polymyxin-acriflavine-lithium chloride-ceftazidime-esculin-mannitol (PALCAM) agar and aerobic plate counts. HPP-induced bacterial injury and its recovery have been investigated by comparing selective and nonselective agar plate counts; however, few investigations have addressed this issue in the presence of essential oil extracts, taking into account the effect of high pressure and natural antimicrobial compounds (e.g., carvacrol) on bacterial survival in various growth media. Use of selective media may overestimate the efficacy of bacterial inactivation in food processing evaluation and validation studies, and the effects of various media should be systematically investigated.

Inactivation of extraintestinal pathogenic E. coli clinical and food isolates suspended in ground chicken meat by gamma radiation.

Extraintestinal pathogenic Escherichia coli are common contaminants in retail poultry and involved inflammatory bowel disease, urinary tract infections and meningitis in both animals and humans. They cause significantly more illnesses and deaths in humans than Shiga toxin-producing E. coli (STEC). Ionizing radiation is used commercially for improving the safety and shelf-life of foods. In this study we inoculated ground chicken meat with 25 individual isolates of clinical uropathogenic E. coli (UPEC) and newborn meningitis causing E. coli (NMEC), isolates from retail chicken meat (CM), as well as retail chicken-skin isolates identified in our laboratory (CS). We then determined their gamma radiation inactivation kinetics (D10-value). The mean D10-value for all isolates (n = 25) was 0.30 kGy. The mean D10-value for the UPEC, NMEC, CM, and CS isolates were 0.25, 0.29, 0.29, and 0.39 kGy, respectively. The mean D10-value for the clinical isolates was 0.27 kGy vs. 0.34 kGy for the non-clinical isolates. There was no correlation between presence of virulence factors, antibiotic resistance, and radiation resistance. ExPEC were similar to that of STEC which were previously evaluated in our laboratory. The radiation doses needed to kill STEC poultry meat should also kill ExPEC.

Draft Genomic Sequence of Escherichia coli Sequence Type 131, Isolated from Retail Chicken Skin.

Escherichia coli sequence type 131 (ST131) is a foodborne pathogen increasingly associated with urinary tract infections. We report here the draft genomic sequence of ST131 B7S75, isolated from retail chicken skin, including information about its virulence factors and antibiotic resistance.

Modeling the Survival of Escherichia coli O157:H7 Under Hydrostatic Pressure, Process Temperature, Time and Allyl Isothiocyanate Stresses in Ground Chicken Meat.

Shiga toxin-producing Escherichia coli O157:H7 (STEC) is a common contaminant in meat and poultry. We investigated the use of non-thermal high pressure processing (HPP), with or without allyl isothiocyanate (AITC) essential oil, to kill STEC in ground chicken meat. Temperature was found an important factor affecting the inactivation of STEC in addition to pressure and process time. A full factorial experiment design (4 factors × 2 levels) was used to facilitate and evaluate the effect of pressure (250-350 MPa), operation temperature (-15-4°C), AITC concentration (0.05-0.15%, w/w), and pressure-holding time (10-20 min) on the inactivation of STEC. A linear model (a polynomial equation) was developed to predict/describe those four parameters' impact on E. coli O157:H7 survival (R2 = 0.90), as well as a dimensionless non-linear model. Both types of models were validated with data obtained from separate experimental points. The dimensionless model also demonstrated that it may predict the lethality (defined as the log CFU/g reduction of STEC before and after treatment) reasonably well with some factors set slightly outside the design ranges (e.g., a wider application than the linear model). The results provide important information regarding STEC survival as affected by HPP (e.g., pressure, time and temperature) and AITC. With the addition of AITC, the hydrostatic pressure may be lowered to the 250-350 MPa level. Regulatory agencies and food industry may use those models for STEC risk assessment in ground chicken meat. A storage test (at 4 and 10°C, 10 days) after HPP+AITC treatment indicated that AITC may continue depressing or killing the pressure-damaged cells.

Draft Genomic Sequencing of Six Potential Extraintestinal Pathogenic Escherichia coli Isolates from Retail Chicken Meat.

Potential extraintestinal pathogenic Escherichia coli strains DP254, WH333, WH398, F356, FEX675, and FEX725 were isolated from retail chicken meat products. Here, we report the draft genome sequences for these six E. coli isolates, which are currently being used in food safety research.

Draft Genome Sequences of Five Neonatal Meningitis-Causing Escherichia coli Isolates (SP-4, SP-5, SP-13, SP-46, and SP-65).

Neonatal meningitis-causing Escherichia coli isolates (SP-4, SP-5, SP-13, SP-46, and SP-65) were recovered between 1989 and 1997 from infants in the Netherlands. Here, we report the draft genome sequences of these five E. coli isolates, which are currently being used to validate food safety processing technologies.

Draft Genome Sequences of Four Uropathogenic Escherichia coli Serotype O4:H5 Isolates (ATCC 700414, 700415, 700416, and 700417).

Uropathogenic Escherichia coli serotype O4:H5 isolates (ATCC 700414, 700415, 700416, and 700417) were recovered from women with first-time urinary tract infections. Here, we report the draft genome sequences for these four E. coli isolates, which are currently being used to validate food safety processing technologies.

Lethality Prediction for Escherichia Coli O157:H7 and Uropathogenic E. coli in Ground Chicken Treated with High Pressure Processing and Trans-Cinnamaldehyde.

Pathogenic Escherichia coli, intestinal (O157:H7) as well as extraintestinal types (for example, Uropathogenic E. coli [UPEC]) are commonly found in many foods including raw chicken meat. The resistance of E. coli O157:H7 to UPEC in chicken meat under the stresses of high hydrostatic Pressure (HHP, also known as HPP-high pressure processing) and trans-cinnamaldehyde (an essential oil) was investigated and compared. UPEC was found slightly less resistant than O157:H7 in our test parameter ranges. With the addition of trans-cinnamaldehyde as an antimicrobial to meat, HPP lethality enhanced both O157:H7 and UPEC inactivation. To facilitate the predictive model development, a central composite design (CCD) was used to assess the 3-parameter effects, that is, pressure (300 to 400 MPa), trans-cinnamaldehyde dose (0.2 to 0.5%, w/w), and pressure-holding time (15 to 25 min), on the inactivation of E. coli O157:H7 and UPEC in ground chicken. Linear models were developed to estimate the lethality of E. coli O157:H7 (R2 = 0.86) and UPEC (R2 = 0.85), as well as dimensionless nonlinear models. All models were validated with data obtained from separated CCD combinations. Because linear models of O157:H7 and UPEC had similar R2 and the significant lethality difference of CCD points was only 9 in 20; all data were combined to generate models to include both O157:H7 and UPEC. The results provide useful information/tool to predict how pathogenic E. coli may survive HPP in the presence of trans-cinnamaldehyde and to achieve a great than 5 log CFU/g reduction in chicken meat. The models may be used for process optimization, product development and to assist the microbial risk assessment. PRACTICAL APPLICATION: The study provided an effective means to reduce the high hydrostatic pressure level with incorporation of antimicrobial compound to achieve a 5-log reduction of pathogenic E. coli without damaging the raw meat quality. The developed models may be used to predict the high pressure processing lethality (and process optimization), product development (ingredient selection), and to assist the microbial risk assessment.

LMOf2365_0442 Encoding for a Fructose Specific PTS Permease IIA May Be Required for Virulence in L. monocytogenes Strain F2365.

Listeria monocytogenes is a foodborne pathogen that causes listeriosis, which is a major public health concern due to the high fatality rate. LMOf2365_0442, 0443, and 0444 encode for fructose-specific EIIABC components of phosphotransferase transport system (PTS) permease that is responsible for sugar transport. In previous studies, in-frame deletion mutants of a putative fructose-specific PTS permease (LMOf2365_0442, 0443, and 0444) were constructed and analyzed. However, the virulence potential of these deletion mutants has not been studied. In this study, two in vitro methods were used to analyze the virulence potential of these L. monocytogenes deletion mutants. First, invasion assays were used to measure the invasion efficiencies to host cells using the human HT-29 cell line. Second, plaque forming assays were used to measure cell-to-cell spread in host cells. Our results showed that the deletion mutant ΔLMOf2365_0442 had reduced invasion and cell-to-cell spread efficiencies in human cell line compared to the parental strain LMOf2365, indicating that LMOf2365_0442 encoding for a fructose specific PTS permease IIA may be required for virulence in L. monocytogenes strain F2365. In addition, the gene expression levels of 15 virulence and stress-related genes were analyzed in the stationary phase cells of the deletion mutants using RT-PCR assays. Virulence-related gene expression levels were elevated in the deletion mutants ΔLMOf2365_0442-0444 compared to the wild type parental strain LMOf2365, indicating the down-regulation of virulence genes by this PTS permease in L. monocytogenes. Finally, stress-related gene clpC expression levels were also increased in all of the deletion mutants, suggesting the involvement of this PTS permease in stress response. Furthermore, these deletion mutants displayed the same pressure tolerance and the same capacity for biofilm formation compared to the wild-type parental strain LMOf2365. In summary, our findings suggest that the LMOf2365_0442 gene can be used as a potential target to develop inhibitors for new therapeutic and pathogen control strategies for public health.

Effects of heat treatment on the antioxidative and anti-inflammatory properties of orange by-products.

This study investigated the changes in the functional components, antioxidative activities, antibacterial activities, anti-inflammatory activities of orange (Citrus sinensis (L.) Osbeck) by-products (OBP) on heat treatment at 50 and 100 °C (hereafter denoted 50D and 100D extracts, respectively). Optimal heating conditions were also investigated. The total phenolic content, flavonoid content and antioxidative activities of OBP extracts significantly increased on heat treatment. The lag time of Cu2+-induced oxidation of human LDL was increased by 2.61, 8.61 and 8.76-fold with the addition of 0.6, 0.8 and 1.0 mg ml-1 100D extracts, respectively. The 100D extracts may significantly inhibit the growth of E. coli O157, Salmonella typhimurium and Listeria monocytogenes. 1 µg mL-1 of 100D extract may suppress the TNF-α-induced ICAM-1 protein expression. The optimal heating time for OBP was 26 h at 100 °C, which resulted in the highest antioxidant activities.

Inactivation of Salmonella spp., pathogenic Escherichia coli, Staphylococcus spp., or Listeria monocytogenes in chicken purge or skin using a 405-nm LED array.

Raw poultry are sometimes contaminated with foodborne pathogens, which can lead to illness in humans. In recent years research has focused on a variety of light technologies to decontaminate food and food contact surfaces during meat and poultry processing. In this study we evaluated the ability of 405-nm light generated from an LED array to inactivate multi-isolate cocktails of either Salmonella spp., pathogenic Escherichia coli, Staphylococcus spp., or Listeria monocytogenes suspended in chicken purge or skin. When exposed to 180 J/cm2 405-nm light at two separate light intensities (300 mW/cm2/s or 150 mW/cm2/s) the maximum pathogen reduction on chicken skin was ca. 0.4 log. When the pathogens were suspended in chicken purge the maximum log reductions ranged from 0.23 to 0.68 log (180 J/cm2; 150 mW/cm2/s) versus 0.69 to 1.01 log (180 J/cm2; 300 mW/cm2/s). Log reductions of each pathogen, when they were subjected to heat shock prior to 405-nm light treatment, were reduced, indicating that thermal effects accounted for much of the bacterial inactivation.