Salmonella and Campylobacter are major foodborne pathogens that cause outbreaks associated with contaminated chicken liver. Proper cooking is necessary to avoid the risk of illness to consumers. This study tested the thermal inactivation of a 4-strain Salmonella cocktail and a 3-strain Campylobacter cocktail in chicken livers separately at temperatures ranging from 55.0 to 62.5°C. Inoculated livers were sealed in aluminum cells and immersed in a water bath. The decimal reduction time (D-values) of Salmonella in chicken livers were 9.01, 2.36, 0.82, and 0.23 min at 55.0, 57.5, 60.0, and 62.5°C, respectively. The D-values of Campylobacter ranged from 2.22 min at 55.0°C to 0.19 min at 60.0°C. Salmonella and Campylobacter had similar z-values in chicken livers of 4.8 and 4.6°C, respectively. Chicken livers can be heated to internal temperatures of 70.0 to 73.9°C for at least 1.6 to 0.2 s to achieve a 7-log reduction of Salmonella. Validation tests demonstrated that heating chicken livers to internal temperatures of 70.0 to 73.9°C for 2 to 0 s resulted in a reduction of Salmonella exceeding 7 logs. Collectively, these data show that Salmonella exhibits higher heat resistance than Campylobacter in chicken livers. Therefore, Salmonella could be considered as the target pathogen when designing thermal treatments or cooking instructions for liver products. These findings will aid in designing effective thermal processing for both industrial and home cooking to eliminate Salmonella and Campylobacter, ensuring consumer safety when consuming chicken liver products.

The objective of this study was to identify a suitable surrogate for E. coli O157:H7 strain 19685/91 and O113:H21 strain TS18/08, by assessing their thermal resistance at temperatures of 60 °C, 65 °C, and 72 °C in strawberry nectar. The influence of the matrix and the research methodology on the decimal reduction time (D-value) was investigated. Thermal kinetics and safety assessment demonstrated that E. coli ATCC 8739 is a suitable surrogate. The study demonstrated that the presence of fruit particles in the nectar increased thermal resistance of the tested strains. Variations in D-values were observed depending on the research method employed, with D-values in glass capillaries were up to 6.6 times lower compared to larger sample volumes. Encapsulation of E. coli ATCC 8739 exhibited high efficiency of 90.25 ± 0.26% and maintained stable viable counts after 26 days of storage in strawberry nectar at 4 °C. There were no significant differences in thermal resistance between surrogates directly inoculated into strawberry nectar and those encapsulated in alginate beads. Additionally, the encapsulated strains did not migrate outside the beads. Therefore, encapsulated E. coli ATCC 8739 in alginate beads can be effectively utilized in industrial settings to validate thermal treatments as a reliable and safe method.

Lytic bacteriophages are promising biocontrol agents against pathogenic bacteria for food and therapeutic applications. Investigating the feasibility of combining phage and physical lethal agents, such as heat, as an effective hurdle combination could lead to beneficial applications. The current research was initiated to compare the thermal inactivation kinetics of a lytic phage (Escherichia phage OSYSP) and its host (Shiga toxin-producing Escherichia coli O157:H7 EDL933), considering they have different critical thermal targets in their structures. To provide a basis for comparison, thermal inactivation kinetics were determined on suspensions of these agents in buffered peptone water using a thermally controlled circulating water bath. Results showed that the bacteriophage virions have a remarkable heat resistance (p < 0.05) compared to their host cells. The D-values of the populations of phage (PFU/mL) and EDL933 strain (CFU/mL) were 166.7 and 7.3 min at 55°C, compared to 44.4 and 0.3 min at 60°C, respectively. Additionally, D-values were significantly (p < 0.05) more influenced by temperature changes in the case of E. coli O157:H7 EDL933 (z-value 3.7°C) compared to that for phage OSYSP (z-value 7.7°C). When the phage suspension was heat-treated in a thermal cycler instead of a water bath, no significant differences between the two treatment procedures (p > 0.05) in estimating virus D- and z-values were observed. Based on these findings, it may be feasible to combine phage OSYSP with mild heat during processing of food to selectively inactivate E. coli O157:H7 EDL933 and subsequently maintain product safety during storage by the surviving phage population; however, the feasibility of this application needs to be investigated. Additionally, the relatively heat-resistant phage OSYSP could qualify as a biological indicator to validate thermal treatments of minimally processed foods in which E. coli O157:H7 EDL933 is the pathogen-of-concern.

Preharvest agricultural water has been recognized as one of the routes of contamination for foodborne pathogens during fruit and vegetable production. Several strategies have been proposed to reduce the risk of pathogens, including preharvest water chemigation, but literature is lacking with regards to microbiological inactivation of common bacterial foodborne pathogens associated with fresh produce contamination, Salmonella enterica, Shiga-toxigenic Escherichia coli (STEC), and Listeria monocytogenes, in surface irrigation water after exposure to chlorine and peracetic acid (PAA). Surface water supplied by a local irrigation district was collected over the summer of 2019. Water was autoclaved, divided into 100 mL samples, and inoculated with a cocktail of five Salmonella, STEC, or Listeria monocytogenes strains or a single strain non-pathogenic E. coli. Samples were then treated with 3, 5, or 7 ppm of free chlorine or PAA, and surviving populations were evaluated using a time-kill assay. A first-order kinetic model was used to fit the inactivation data and obtain the D-values. A secondary model was used to explain the changes due to the type of water, treatment, and microorganism. At a concentration of 3 ppm, the observed and predicted D-values of free chlorine treatments were higher than that of PAA treatments for ground and surface water. Results indicated that PAA was more effective inactivating bacteria than sodium hypochlorite at concentrations of 3 and 5 ppm for both water sources (surface and ground). However, at 7 ppm, the effectiveness of PAA and sodium hypochlorite showed no statistically significant difference for both surface and groundwater. Findings will provide information regarding efficacy of chemical sanitizers like chlorine and PAA for inactivation of Salmonella, Listeria, and STEC in surface water from which treatments can be derived. Ultimately benefitting growers in the selection of an appropriate method for in-field treatment of irrigation water if deemed necessary.

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.

Chicken is a suspected reservoir of uropathogenic Escherichia coli (UPEC), resulting in foodborne urinary tract infections (UTIs). Sous-vide ready-to-eat (RTE) food products may be associated with microbial hazards due to the low-temperature long-time (LTLT) process. However, little is known regarding the survival of UPEC during sous-vide cooking. The aim of this study was to evaluate the heat resistance of UPEC in chicken breast during sous-vide processing and establish predictive inactivation models. Chicken breast samples were inoculated with a four-strain cocktail of UPEC, including reference strains from UTI patients and chicken isolates. The inoculated samples, with or without 3% NaCl solution for marination, were vacuum sealed in bags, immersed in a temperature-controlled water bath, and cooked at 50 °C, 55 °C, 60 °C, and 63 °C. The change in survival of populations of UPEC was fitted with the linear and Weibull inactivation models to obtain the survival curves at different temperatures; the D- and z-values were also calculated. The goodness-of-fit was evaluated using the root mean square error (RMSE), sum of squared errors (SSE), adjusted R2, and Akaike information criterion (AIC). The results showed that the linear model with tail was better than the Weibull model in terms of fitting performance. With the addition of salt marinade, D-values at 50 °C, 55 °C, 60 °C, and 63 °C determined by the linear model with tail decreased from 299.78 to 166.93 min, 16,60 to 13.87 min, 4.06 to 3.05 min, and 1.05 to 0.87 min, respectively, compared with the controls. The z-values of control and salt-marinated samples were 6.14 °C and 5.89 °C, respectively. The model developed for predicting UPEC survival under sous-vide cooking was validated using an additional survival curve at 58 °C. The validation results showed that the RMSE was 0.122 and 0.133 log CFU/g, and the proportion of relative error was 0.875 and 0.750 in the acceptable prediction zones for the control and salt-marinated samples, respectively. In conclusion, the heat resistance of an emerging foodborne pathogen, UPEC, in sous-vide processed chicken breast was revealed for the first time. Our results showed that salt marinade (3% NaCl) increases the heat sensitivity of UPEC during the sous-vide processing. The developed survival functions based on the linear model with tail can be applied to control the thermal lethality of UPEC.

Adequate surrogate identification is critical for validating in-plant thermal process controls for Salmonella inactivation in different food matrices. This study compared the thermal inactivation parameters (D- and z-values) and evaluated the heat resistance of Enterococcus faecium (8459) as a surrogate for a 5-serovar Salmonella cocktail in cornmeal. The cornmeal was spray inoculated with the respective bacteria to achieve ~9 log CFU/g population and set to the desired moisture contents (16, 22, and 28% w.b.). The inoculated cornmeal was then heat-treated at pre-determined temperatures (60, 64, and 68 °C) in sealed aluminum thermal-death-time disks in hot water baths for pre-determined time intervals. Injury-recovery media [brain heart infusion (BHI) agar overlaid with xylose lysine deoxycholate (XLD) agar for Salmonella or BHI agar overlaid m-enterococcus agar for E. faecium] were used for microbial enumeration to account for thermally injured bacterial cells. The D-values of Salmonella in cornmeal at 16, 22, and 28% moisture content were 37.5, 8.4, and 2.4 min at 60 °C, 19.9, 3.5, and 1.1 min at 64 °C, and 10.1, 1.4, and 0.5 min at 68 °C, respectively. The D-values of E. faecium in cornmeal at 16, 22, and 28% moisture content were 140.4, 18.9, and 3.3 min at 60 °C, 78.4, 7.1, and 1.6 min at 64 °C, and 37.3, 2.8, and 0.8 min at 68 °C, respectively. The z-values of E. faecium and Salmonella in cornmeal at 16, 22, and 28% moisture content were 13.9, 9.7, and 12.5 °C, and 14.0, 10.4, and 11.7 °C, respectively. These results indicated similar or higher thermal resistance (D-values) and equivalent thermal sensitivity (z-values) of E. faecium compared to Salmonella at different moisture contents and respective temperatures (P ≤ 0.05). Therefore, E. faecium could be used as a surrogate for Salmonella during thermal process validation of cornmeal processing.

The objective of this work is to match available phylogenetic information for Bacillus cereus strains with published thermal resistance parameters (D90°C, z) and to use this information to develop refined inactivation models for B. cereus sensu lato. To do so, the thermal resistance parameters were retrieved for 57 strains of B. cereus that could be assigned to a phylogenetic group. This information was used to build specific distributions for D90°C and z for the different phylogenetic groups of B. cereus to build refined thermal inactivation models for B. cereus. For validation purposes, thermal parameters were also retrieved for additional strains of unknown groups, but which had been classified as psychrotrophic or mesophilic. Monte Carlo simulations were first performed assuming that the model parameters D90°C and z are independent. However, based on the observation that combinations of very high D90°C and high z-values were not reported, an alternative Monte Carlo simulation set was explored for the phylogenetic Groups with very high z-values (i.e.i.e. Groups IV and VI). With both simulation sets, the predicted lower and upper limits of the D-values are close to the lowest and highest D-values reported in two previous meta-analysis studies. However, a better correspondence between the predicted and observed limits is obtained when using the alternative simulation set.

Hummus is a popular dip in the Middle East region prepared by mixing the boiled chickpeas with tahini and other ingredients, and because its consumption has increased world-wide some notoriety has developed following an increase in the incidence of hummus-related illness outbreaks and recalls. The objectives of the current research were (i) to study the efficiency of low dose gamma irradiation to inhibit Escherichia coli O157:H7, Salmonella enterica, and Listeria monocytogenes in hummus, and (ii) to assess the effect of environmental stresses namely cold, heat, and desiccation on the resistance of these pathogens to gamma irradiation. The samples of hummus were prepared and then individually inoculated with approximately 7.0 log CFU/g of unstressed or cold-, heat-, or desiccated-stressed cocktail cultures of each of E. coli O157:H7, S. enterica, and L. monocytogenes. The inoculated samples were then exposed to gamma irradiation at doses of 0.1 to 0.6 kGy. The numbers of unstressed E. coli O157:H7, S. enterica, and L. monocytogenes were decreased by 0.6-3.9, 0.7-2.9, and 1.0-3.0 log CFU/g, respectively, by irradiation treatment. The resistance of E. coli O157:H7 to gamma irradiation was not affected by desiccation, heat, and cold stresses. However, the pre-exposure of S. enterica and L. monocytogenes cells to these stresses reduced their resistance toward gamma irradiation. PRACTICAL APPLICATION: Gamma irradiation is a non-thermal treatment that can be used in food processing to ensure food safety and quality. The current study proved that low levels (≤0.6 kGy) of gamma irradiation can effectively decrease the risk of unstressed and cold-, heat-, or desiccation-stressed Salmonella enterica, Listeria monocytogenes, or Escherichia coli O157:H7 in hummus.

Bacillus cereus, a foodborne pathogen, is capable of forming spores and biofilms as methods to withstand environmental stresses. These bacterial structures are an issue for food safety as they aid the bacteria survive heat sterilisation processes of foods and food contact surfaces. This study was conducted to investigate the role of the biofilm structure in providing an extra layer of protection to spores against heat treatments. For this, heat resistance of B. cereus spores in intact biofilms was compared to that of planktonic spores in vitro and in a Cheonggukjang jjigae food model. Using methods developed in this study to measure the wet and dry heat resistance of spores in intact biofilms, it was found that B. cereus spores have significantly higher heat resistances when present in biofilms rather than as planktonic spores, and that dry heat is less effective than wet heat at killing spores in biofilms. In further detail, for wet heat treatments, spores in biofilms of the strain isolated from Cheonggukjang (Korean fermented whole soybean), B. cereus CH3, had generally higher wet heat resistances than the reference strain, B. cereus ATCC 10987, both in vitro and in the Cheonggukjang jjigae food model. However, the spores in biofilms of the two strains showed similar heat resistance to dry heat, with some exceptions, when biofilms were formed in vitro or in Cheonggukjang jjigae broth. Meanwhile, B. cereus ATCC 10987 spores in biofilms had higher or similar wet heat resistances in vitro compared to in Cheonggukjang jjigae broth. Wet heat resistances of B. cereus CH3 spores in biofilms were all statistically similar regardless of biofilm formation media (brain heart infusion and Cheonggukjang jjigae broths). For dry heat, spores in biofilms of both B. cereus strains were more heat resistant when biofilms were formed in the Cheonggukjang jjigae food model rather than in vitro. Altogether, heat resistances of spores in biofilms formed in vitro and in the food environment were found to be different depending on the tested B. cereus strain, but higher than planktonic spores in any case. This is the first study examining the heat resistance of B. cereus spores in intact biofilms matrices attached to the surface, both in vitro and in a food model. Therefore, this research is valuable to understand the protective effects of biofilms formed in food environments and to reduce the food safety risks associated with B. cereus.