Emergent methods for inactivation of Cronobacter sakazakii in foods: A systematic review and meta-analysis.

Cronobacter sakazakii is a potentially pathogenic bacterium that is resistant to osmotic stress and low aw, and capable of persisting in a desiccated state in powdered infant milks. It is widespread in the environment and present in various products. Despite the low incidence of cases, its high mortality rates of 40 to 80 % amongst neonates make it a microorganism of public health interest. This current study performed a comparative assessment between current reduction methods applied for C. sakazakii in various food matrices, indicating tendencies and relevant parameters for process optimization. A systematic review and meta-analysis were conducted, qualitatively identifying the main methods of inactivation and control, and quantitatively evaluating the effect of treatment factors on the reduction response. Hierarchical clustering dendrograms led to conclusions on the efficiency of each treatment. Review of recent research trend identified a focus on the potential use of alternative treatments, with most studies related to non-thermal methods and dairy products. Using random-effects meta-analysis, a summary effect-size of 4-log was estimated; however, thermal methods and treatments on dairy matrices displayed wider dispersions - of τ2 = 8.1, compared with τ2 = 4.5 for vegetal matrices and τ2 = 4.0 for biofilms. Meta-analytical models indicated that factors such as chemical concentration, energy applied, and treatment time had a more significant impact on reduction than the increase in temperature. Non-thermal treatments, synergically associated with heat, and treatments on dairy matrices were found to be the most efficient.

Evaluation of air impingement for dry-cleaning nonfat dry milk residues on a stainless-steel surface.

The use of air jet impingement to remove residues from surfaces in food manufacturing operations offers an alternative to the use of water and liquid cleaning agents. During this investigation, air impingement was used to remove nonfat dry milk (NFDM) residues from a stainless-steel surface. The influence of the water activity (aw ) of the residue, the time after the residue reached an equilibrium water activity, and the thickness of residue at the time of removal from the surface have been investigated. All three factors had a significant effect on the time for removal. An increase in the water activity, the time at equilibrium, the sample thickness, or a combination of all three resulted in an increase in the time required to remove the deposits. Visible changes in the structure of deposits were observed as NFDM samples equilibrated to water activities above 0.43. NFDM residues with water activities less than 0.33 were removed within 1 s of using air impingement regardless of wall shear stress. When the water activities were greater than 0.50, the thickness of deposit was greater than 1 mm, and the time after reaching an equilibrium water activity was over 7 days, more than 5 min of air impingement with wall shear stress over 9.48 Pa was required to remove the residue. The results from these experiments indicated that air impingement has the potential to provide effective cleaning in manufacturing facilities for low-moisture foods. PRACTICAL APPLICATION: The introduction of water in low-moisture food environments is often undesirable due to the possibility of pathogenic microorganism growth. The normal cleaning operations in the food industry use water as a cleaning agent. This study evaluates the application of air impingement technology as a dry-cleaning method.

Effectiveness of Ultra-High Irradiance Blue-Light-Emitting Diodes to Control Salmonella Contamination Adhered to Dry Stainless Steel Surfaces.

Controlling Salmonella contamination in dry food processing environments represents a significant challenge due to their tolerance to desiccation stress and enhanced thermal resistance. Blue light is emerging as a safer alternative to UV irradiation for surface decontamination. In the present study, the antimicrobial efficacy of ultra-high irradiance (UHI) blue light, generated by light-emitting diodes (LEDs) at wavelengths of 405 nm (841.6 mW/cm2) and 460 nm (614.9 mW/cm2), was evaluated against a five-serovar cocktail of Salmonella enterica dry cells on clean and soiled stainless steel (SS) surfaces. Inoculated coupons were subjected to blue light irradiation treatments at equivalent energy doses ranging from 221 to 1106 J/cm2. Wheat flour was used as a model food soil system. To determine the bactericidal mechanisms of blue light, the intracellular concentration of reactive oxygen species (ROS) in Salmonella cells and the temperature changes on SS surfaces were also measured. The treatment energy dose had a significant effect on Salmonella inactivation levels. On clean SS surfaces, the reduction in Salmonella counts ranged from 0.8 to 7.4 log CFU/cm2, while, on soiled coupons, the inactivation levels varied from 1.2 to 4.2 log CFU/cm2. Blue LED treatments triggered a significant generation of ROS within Salmonella cells, as well as a substantial temperature increase in SS surfaces. However, in the presence of organic matter, the oxidative stress in Salmonella cells declined significantly, and treatments with higher energy doses (>700 J/cm2) were required to uphold the antimicrobial effectiveness observed on clean SS. The mechanism of the bactericidal effect of UHI blue LED treatments is likely to be a combination of photothermal and photochemical effects. These results indicate that LEDs emitting UHI blue light could represent a novel cost- and time-effective alternative for controlling microbial contamination in dry food processing environments.

Evaluation of the Handling Practices and Risk Perceptions of Dried Wood Ear Mushrooms in Asian Restaurants in the United States.

In late 2020, dried wood ear mushrooms, a low-moisture food ingredient that had been imported and sold to restaurants, were linked to a foodborne outbreak of Salmonella Stanley, which sickened 55 individuals across the United States. These mushrooms are commonly used in Asian cuisine. It is unclear if the contaminated dried wood ear mushrooms that caused the foodborne illnesses were improperly handled during preparation. The objectives of this study are to assess the handling practices, risk perceptions, and food recall experiences of dried wood ear mushrooms in restaurant kitchens among Asian restaurant managers and chefs. We conducted a series of telephone interviews with managers and chefs of Asian restaurants in the United States who used dried wood ear mushrooms in making dishes. After reaching information saturation, a total of 25 restaurant managers and chefs participated in the interview. Our results showed that 76% of the participants did not keep track of package information, such as expiration date and lot number, and many participants reported using cold water for rehydration. Wood ear mushrooms were blanched before being used in all cold dishes and most stir-fry dishes, but less commonly in stew or ramen. Some participants (16%) did not view dried wood ear mushrooms as a raw food ingredient, and 16% did not perceive that low-moisture food ingredients constituted microbiological food safety risks. The majority of the participants had heard of food recalls, but only 17% knew about the dried wood ear mushroom recall, and even fewer had heard of food recalls of other low-moisture foods, like nuts and seeds (9%), and flour (4%). While this study shares similarities with previously published studies evaluating the handling practices of consumers and restaurant employees with respect to meat and poultry, it makes a distinctive contribution to the field of food safety as the first-of-its-kind to study the handling practices of a low-moisture food ingredient: dried wood ear mushrooms. This unique ethnic food ingredient has been associated with a past outbreak and multiple recalls in the United States. The findings of the study show the need to develop food safety educational programs that are tailored toward Asian restaurant food handlers and provide guidance to develop risk communication strategies for this niche audience.

Effectiveness of Ultra-High Irradiance Blue Light-Emitting Diodes in Inactivating Escherichia coli O157:H7 on Dry Stainless Steel and Cast-Iron Surfaces.

The use of blue light-emitting diodes (LEDs) is emerging as a promising dry decontamination method. In the present study, LEDs emitting ultra-high irradiance (UHI) density at 405 nm (842 mW/cm2) and 460 nm (615 mW/cm2) were used to deliver high-intensity photoinactivation treatments ranging from 221 to 1107 J/cm2. The efficacy of these treatments to inactivate E. coli O157:H7 dry cells was evaluated on clean and soiled stainless steel and cast-iron surfaces. On clean metal surfaces, the 405 and 460 nm LED treatment with a 221 J/cm2 dose resulted in E. coli reductions ranging from 2.0 to 4.1 log CFU/cm2. Increasing the treatment energy dose to 665 J/cm2 caused further significant reductions (>8 log CFU/cm2) in the E. coli population. LED treatments triggered a significant production of intracellular reactive oxygen species (ROS) in E. coli cells, as well as a significant temperature increase on metal surfaces. In the presence of organic matter, intracellular ROS generation in E. coli cells dropped significantly, and treatments with higher energy doses (>700 J/cm2) were required to uphold antimicrobial effectiveness. The mechanism of the bactericidal effect of UHI blue LED treatments is likely to be a combination of photothermal and photochemical effects. This study showed that LEDs emitting monochromatic blue light at UHI levels may serve as a viable and time-effective method for surface decontamination in dry food processing environments.

U.S. Consumer Practices of Homemade Nut-based Dairy Analogs and Soaked Nuts.

Tree nuts, a low-moisture food, are typically perceived as being a low risk for foodborne illness. In the past five decades, the consumption of tree nuts (dry, soaked, or as nut-based dairy analogs [NBDA]) has increased along with corresponding foodborne illness outbreaks and recalls associated with these products. We developed an online survey to assess tree nut handling practices of U.S. consumers, and to select study participants who have soaked tree nuts and/or made NBDA at home. We distributed our initial survey questions in October 2021 to a convenience sample (n = 12) to test for clarity and comprehension. In January 2022, participants (n = 981) who met the criteria completed the survey. The most popular soaked tree nuts were almonds (54%), followed by cashews (36%), walnuts (32%), and pistachios (22%). Participants soaked tree nuts for direct consumption (67%) and during the preparation of NBDA (80%). Participants soaked tree nuts under refrigerated conditions for 1-24 h (22%), on the countertop at room temperature (est. 65-75°F [18-24°C]) for 1-5 h (21%), or at room temperature for 12 h or more (6%); 16% used a hot or boiling water, short time treatment. Some participants added acid (28%) or salt (25%) to the soaking water. Among those participants who dried their tree nuts after soaking (63%), 89% reported drying at a temperature lower than 46°C (115°F). Some participants (34%) used their tree nuts to make fermented dairy analogs (e.g., "cheese" or "yogurt") by adding "probiotics" (56-86%) or a yogurt starter culture (37-99%), respectively, and then, most frequently, holding at or below 20°C (68°F) for 12 h or less (29%). The safety of many of these practices has not been adequately investigated, but the findings of this study will inform future risk assessment and risk modeling studies on tree nut food safety in home kitchen settings.

Mild heat treatment achieved better inactivation of Salmonella and preservation of almond quality than ultraviolet light and chemical sanitizers.

This study was conducted to compare the effects of ultraviolet light (UV), chemical sanitizers, and heat treatments on Salmonella inactivation and preservation of almond quality. Whole, skinless, and sliced almonds, representing different shape and surface topography, were inoculated with a Salmonella cocktail consisting of S. Montevideo, S. Newport, S. Typhimurium, S. Heidelberg, and S. Enteritidis. Inoculated almonds (50 g) were treated by UV (30 mW/cm2, 30 or 60 min), 75 °C heat (up to 150 min), and chemical sanitizers (3 % hydrogen peroxide (H2O2) and 1 % cetylpyridinium chloride (CPC), 30 or 60 min) alone or in combinations. Uninoculated almonds were similarly treated for analyzing color, visual appearance, and weight changes. In general, UV treatment alone was ineffective in inactivating Salmonella; the 30- and 60-min UV treatments reduced Salmonella by 1.3 (± 0.1) and 1.7 (± 0.1), 2.7 (± 0.2) and 3.3 (± 0.1), and 1.3 (± 0.1) and 1.7 (± 0.1) log CFU/g on whole, skinless, and sliced almonds, respectively. Prior wetting of almonds with water and chemical solutions in a few cases significantly (P < 0.05) increased the UV inactivation of Salmonella. The most pronounced Salmonella killing effect achieved by the combined treatments were: 1-min H2O2 dipping followed by 60-min UV treatment for whole (3.0 logs) and skinless almonds (3.8 logs) and 1-min CPC dipping followed by 60-min UV treatment for sliced almonds (3.0 logs). However, none of those achieved >4 log reductions of Salmonella as required by FDA. The 30-min UV treatment produced discolored but overall acceptable almonds, whereas the 60-min UV treatment led to deteriorated almonds including a dark color, oil extraction, and shrunk kernel size. Prior wetting reduced the sample weight loss but caused local burning and kernel cracking. A sequential approach of a 60-min 75 °C heat treatment and two 30-min wet UV treatments successfully reduced Salmonella by >4 logs, but more severe kernel cracking occurred. In contrast, a single heat treatment of vacuum packaged whole almonds at 75 °C for 150 min was capable of achieving >5 log reductions of Salmonella while preserving almond color and visual qualities and minimizing weight loss. These results clearly demonstrated that the heat treatment was a much better processing technology than UV and sanitizers for raw almond pasteurization.

Generation, mechanisms, kinetics, and effects of gaseous chlorine dioxide in food preservation.

Food preservation is a critical issue in ensuring food safety and quality. Growing concern around industrial pollution of food and demand for environmentally sustainable food has led to increased interest in developing effective and eco-friendly preservation techniques. Gaseous ClO2 has gained attention for its strong oxidizing properties, high efficacy in microorganism inactivation, and potential for preserving the attributes and nutritional quality of fresh food while avoiding the formation of toxic byproducts or unacceptable levels of residues. However, the widespread use of gaseous ClO2 in the food industry is limited by several challenges. These include large-scale generation, high cost and environmental considerations, a lack of understanding of its mechanism of action, and the need for mathematical models to predict inactivation kinetics. This review aims to provide an overview of the up-to-date research and application of gaseous ClO2 . It covers preparation methods, preservation mechanisms, and kinetic models that predict the sterilizing efficacy of gaseous ClO2 under different conditions. The impacts of gaseous ClO2 on the quality attributes of fresh produce and low-moisture foods, such as seeds, sprouts, and spices, are also summarized. Overall, gaseous ClO2 is a promising preservation approach, and future studies are needed to address the challenges in large-scale generation and environmental considerations and to develop standardized protocols and databases for safe and effective use in the food industry.

The influence of almond's water activity and storage temperature on Salmonella survival and thermal resistance.

This study investigated the effects of inoculation method, water activity (aw), packaging method, and storage temperature and duration on the survival of Salmonella on almonds as well as their resistance to subsequent thermal treatments. Whole almond kernels were inoculated with a broth-based or agar-based growth Salmonella cocktail and conditioned to aw of 0.52, 0.43 or 0.27. Inoculated almonds with aw of 0.43 were treated with a previously validated treatment (4 h of dry heat at 73 °C) to determine the potential differences in heat resistance resulted from the two inoculation methods. The inoculation method did not significantly (P > 0.05) impact the thermal resistance of Salmonella. Inoculated almonds at aw of 0.52 and 0.27 were either vacuum packaged in moisture-impermeable mylar bags or non-vacuum packaged in moisture-permeable polyethylene bags before stored at 35, 22, 4, or -18 °C for up to 28 days. At selected storage intervals, almonds were measured for aw, analyzed for Salmonella population level, and subjected to dry heat treatment at 75 °C. Over the month-long storage of almonds, Salmonella populations remained almost unchanged (<0.2 log CFU/g) at 4 °C and -18 °C and declined slightly (<0.8 log CFU/g) at 22 °C and more substantially (1.6-2.0 log CFU/g) at 35 °C regardless of the inoculation method, packaging method, and almond aw. When stored at 35 °C, almonds with initial aw of 0.52 had significantly higher (P < 0.05) Salmonella reductions than those with initial aw of 0.27. Prior storage of almonds vacuum packaged in mylar bags at temperatures between -18 °C and 35 °C for 28 days affected their aw levels but did not significantly (P > 0.05) affect the subsequent thermal resistance of Salmonella at 75 °C regardless of almond aw and storage duration. Salmonella on almonds with higher aw was more sensitive to heat treatment than those with lower aw. To achieve >5 log CFU/g reductions of Salmonella, a dry heat treatment at 75 °C for 4 and 6 h was needed for almonds with initial aw of 0.52 and 0.27, respectively. When applying the dry heating technology for almond decontamination, the processing time needs to be determined based on initial aw of almonds regardless of storage condition or age of almonds within the current design frame.

Effect of oil exposure stages on the heat resistance of Salmonella enterica serovar Enteritidis phage type 30 in peanut flour.

The oil in low-moisture foods (LMFs) shows protective effects on bacteria during thermal processing. However, the circumstances under which this protective effect strengthens remain unclear. This study aimed to understand which step of the oil exposure to bacterial cells (inoculation, isothermal inactivation, or recovery and enumeration step) in LMFs can enhance their heat resistance. Peanut flour (PF) and defatted PF (DPF) were selected as the oil-rich and oil-free LMF models. Salmonella enterica Enteritidis Phage Type 30 (S. Enteritidis) was inoculated into four designated PF groups representing different oil exposure stages. It was isothermally treated to obtain heat resistance parameters. At a constant moisture content (aw,25°C = 0.32 ± 0.02) and controlled aw,85°C (0.32 ± 0.02), S. Enteritidis exhibited significantly high (p < 0.05) D values in oil-rich sample groups. For instance, the heat resistance values of S. Enteritidis in the PF-DPF and DPF-PF groups were D80°C of 138.22 ± 7.45 min and 101.89 ± 7.82 min; however, the D80°C in the DPF-DPF group was 34.54 ± 2.07 min. The oil addition after the thermal treatment also helped injured bacterial recovery in the enumeration. For instance, the D80°C, D85°C, and D90°C values in the DFF-DPF oil groups were 36.86 ± 2.30, 20.65 ± 1.23, and 7.91 ± 0.52 min, respectively, which were higher than those in the DPF-DPF group at 34.54 ± 2.07, 17.87 ± 0.78, and 7.10 ± 0.52 min. We confirmed that the oil protected S. Enteritidis in PF in all three stages: desiccation process, heat treatment, and recovery of bacterial cells in plates.

Oil-Based Sanitization in Low-Moisture Environments: Delivery of Acetic Acid with Water-in-Oil Emulsions.

Contamination with Salmonella spp. and Listeria monocytogenes is concerning across low-moisture food (LMF)-processing environments due to the pronounced survival of these organisms under dry conditions. This study treated desiccated bacteria with acetic acid delivered by oil with and without water-in-oil (W/O) emulsion. The influences of cellular desiccation, emulsion water concentration, water activity (aw), and treatment temperature were investigated. Acetic acid dissolved in oil (i.e., acidified oil) showed low levels of antimicrobial efficacy. After treatment with acidified oil (200 mM acetic acid at 22°C for 30 min), Salmonella enterica serovar Enteritidis phage type 30 cells desiccated to 75% equilibrium relative humidity (ERH) and 33% ERH were reduced by 0.69 and 0.05 log CFU/coupon, respectively. The dispersion of a low level of water (≥0.3%, vol/vol) within the acidified oil with the surfactant (i.e., acidified W/O emulsion) significantly enhanced the antimicrobial efficacy. After treatment with the acidified W/O emulsion (200 mM acetic acid at 22°C for 20 min), desiccated Salmonella (4-strain cocktail) and L. monocytogenes (3-strain cocktail) cells were reduced by >6.52 log most probable number (MPN)/coupon, regardless of the desiccation levels. Increased efficacy was observed with temperature elevation. Reduced efficacy was observed when glycerol was added to the aqueous phase of the emulsion to decrease the solution aw, indicating that the enhanced efficacy of the acidified W/O emulsion was associated with differential osmotic pressure. The antimicrobial mechanism may be due to the membrane disruption induced by acetic acid, in combination with the hypoosmotic stress provided by W/O emulsion, creating cellular lysis, as illustrated by electron micrographs. IMPORTANCE Aqueous-based cleaning and sanitation are undesirable in processing facilities that manufacture low-moisture foods such as peanut butter and chocolate. Alcohol-based sanitization is advantageous because it leaves no residue on the contact surface but requires the processing facility to close temporarily due to flammability. At >6.52 log kill of desiccated Salmonella and Listeria monocytogenes cells, the developed oil-based formulation has the potential to be an effective dry sanitation method.

Hurdle Technology Approach to Control Listeria monocytogenes Using Rhamnolipid Biosurfactant.

This study evaluates the combination of mild heat with a natural surfactant for the inactivation of L. monocytogenes Scott A in low-water-activity (aw) model systems. Glycerol or NaCl was used to reduce the aw to 0.92, and different concentrations of rhamnolipid (RL) biosurfactant were added before heat treatment (60 °C, 5 min). Using glycerol, RL treatment (50-250 µg/mL) reduced bacterial population by less than 0.2 log and heat treatment up to 1.5 log, while the combination of both hurdles reached around 5.0 log reduction. In the NaCl medium, RL treatment displayed higher inactivation than in the glycerol medium at the same aw level and a larger synergistic lethal effect when combined with heat, achieving ≥ 6.0 log reduction at 10-250 µg/mL RL concentrations. The growth inhibition activity of RL was enhanced by the presence of the monovalent salts NaCl and KCl, reducing MIC values from >2500 µg/mL (without salt) to 39 µg/mL (with 7.5% salt). The enhanced antimicrobial activity of RL promoted by the presence of salts was shown to be pH-dependent and more effective under neutral conditions. Overall, results demonstrate that RL can be exploited to design novel strategies based on hurdle approaches aiming to control L. monocytogenes.

Salmonella enterica Outbreaks Linked to the Consumption of Tahini and Tahini-Based Products

Salmonella is a leading cause of bacterial foodborne illness in the world. Although typically associated with foods of animal origin, low-moisture foods, such as tahini, are quickly gaining recognition as an important vehicle of Salmonella exposure. This review offers the Canadian perspective on the issue of Salmonella in tahini and tahini-based products. A summary of several recent food product recalls and foodborne outbreaks related to the presence of Salmonella in tahini and tahini-based products such as halva are presented. The properties of the food vehicles, their production practices, and potential routes of contamination are discussed. Particular focus is placed on the ecology of Salmonella in the tahini production continuum, including its survival characteristics and response to intervention technologies.

Thermal Inactivation of Salmonella enterica and Nonpathogenic Bacterial Surrogates in Wheat Flour by Baking in a Household Oven.

ABSTRACT: Wheat flour has been implicated in recalls and outbreaks linked to Salmonella and pathogenic Escherichia coli. An instructional online video posted on a popular YouTube channel with over 20 million subscribers claimed that safe raw cookie dough could be made from flour baked in a household oven at 177°C (350°F) for 5 min, but no evidence in support of that claim was provided. This study was conducted to assess thermal inactivation of two Salmonella strains, as well as Enterobacter aerogenes and Pantoea dispersa in wheat flour during home oven baking. Wheat flour was inoculated with Salmonella Enteritidis PT 30, Salmonella Typhimurium PT 42, or their potential surrogates at high concentrations (4.8 to 6.1 log CFU/g) before baking in a consumer-style convection oven (toaster oven) at 149, 177, and 204°C (300, 350, or 400°F) for up to 7 min. Flour was heated in an aluminum tray, with a maximum depth of ∼2 cm. Heated wheat flour samples (5 g each) were enumerated in triplicate, and the microbial concentration was expressed in log CFU per gram. Thermal profiles of the geometric center of the wheat flour pile and air in the oven during the baking were recorded. Water activity of wheat flour samples was also measured before and after baking. The water activity of wheat flour decreased, as baking temperature and time increased. Water activity values ranged from 0.30 to 0.06 after 7 min, as oven temperature increased from 149 to 204°C. Thermal inactivation kinetics were linear until counts approached the limit of detection for all microorganisms. D-values for Salmonella and potential surrogate strains ranged from 1.86 to 2.13 min at 149°C air temperature, 1.66 to 1.92 min at 177°C air temperature, and 1.12 to 1.38 min at 204°C air temperature. Both Salmonella strains and surrogates showed similar inactivation patterns. Baking of wheat flour in household toaster ovens has potential as an inactivation treatment of pathogenic bacteria in consumer homes, despite its low water activity.

Sporulation and Biofilms as Survival Mechanisms of Bacillus Species in Low-Moisture Food Production Environments.

Low-moisture foods (LMF) have clear advantages with respect to limiting the growth of foodborne pathogens. However, the incidences of Bacillus species in LMF reported in recent years raise concerns about food quality and safety, particularly when these foods are used as ingredients in more complex higher moisture products. This literature review describes the interlinked pathways of sporulation and biofilm formation by Bacillus species and their underlying molecular mechanisms that contribute to the bacteriums' persistence in LMF production environments. The long-standing challenges of food safety and quality in the LMF industry are also discussed with a focus on the bakery industry.

Evaluation of Potential for Butyl and Heptyl Para-Hydroxybenzoate Enhancement of Thermal Inactivation of Cronobacter sakazakii during Rehydration of Powdered Infant Formula and Nonfat Dry Milk.

ABSTRACT: In previous studies, parabens in model systems enhanced the thermal inactivation of foodborne pathogens, including Cronobacter sakazakii, Salmonella enterica serotype Typhimurium, Escherichia coli O157:H7, and Listeria monocytogenes. However, few studies have been conducted to evaluate this phenomenon in actual food systems. In the present study, the potential enhancement of thermal inactivation of C. sakazakii by butyl para-hydroxybenzoate (BPB) was evaluated in powdered infant formula (PIF) and nonfat dry milk (NFDM) in dry and rehydrated forms. When PIF was rehydrated with water at designated temperatures (65 to 80°C) in baby bottles, BPB did not enhance thermal inactivation. When rehydrated NFDM and lactose solutions with BPB were inoculated and heated at 58°C, BPB enhancement of thermal inactivation of C. sakazakii was negatively associated with the concentration of NFDM solutions in a dose-dependent manner, whereas thermal inactivation was enhanced in the presence of lactose regardless of its concentration, suggesting an interaction between proteins and BPB. Fluorescence testing further indicated an interaction between BPB and the proteins in PIF and NFDM. In inoculated dry NFDM with and without BPB stored at 24 and 55°C for 14 days, BPB did not substantially enhance bacterial inactivation. This study suggests that BPB is not likely to enhance mild thermal bacterial inactivation treatments in foods that have appreciable amounts of protein.

Modeling the effect of protein and fat on the thermal resistance of Salmonella enterica Enteritidis PT 30 in egg powders.

Microorganisms in low-moisture foods (LMFs) exhibit prolonged survivability and high heat resistance. Various external factors (water, food texture, nutritional compounds, etc.) influence the microbial heat resistance in LMFs; yet, the influential degree of each factor is not fully understood. In this study, the thermal resistance parameters (D and z values) of Salmonella enterica Enteritidis PT 30 (S. Enteritidis) at 80, 85, and 90 °C at the room-temperature water activity (aw, 25°C) of 0.32 ± 0.02 were measured. A series of egg powders with different fat and protein ratios (obtained by mixing egg white and yolk powders) were chosen as the model foods. Primary and secondary models were built from the isothermal inactivation kinetics of S. Enteritidis in the tested samples. The importance of fat and protein was then confirmed by controlling the water activity at the treatment temperature (aw, treatment temperature) via thermal water activity cells. The survivor curves of S. Enteritidis fitted well with the Weibull-type and log-linear models. The D values of S. Enteritidis increased with increasing fat (0-56.7%, w.b.) and decreasing protein contents (83.59-31.81%, w.b.). Incorporating the modified Bigelow model into the log-linear model yielded the zfat and zprotein of 58.96 and 57.14, respectively. At the controlled aw, 90°C of 0.32 ± 0.02, the D90°C values of S. Enteritidis increased remarkably (P < 0.05), but the values in egg white, whole egg, and egg yolk powders (11.73 ± 1.24, 23.82 ± 2.0, and 60.0 ± 2.4 min) were remarkably different. Our study identified that the influential degrees of fat, protein (zfat and zprotein values), and aw on the thermal resistance of S. Enteritidis in egg powders is in the order: aw,treatment temperature > fat > protein. Fat considerably increased the thermal resistance of S. Enteritidis even at the same aw,treatment temperature. This study quantified the effect of fat and protein on the thermal resistance of S. Enteritidis and emphasized the non-negligible effects of food components in LMFs' microbial safety.

Survival kinetics, membrane integrity and metabolic activity of Salmonella enterica in conventionally and osmotically dehydrated coconut flakes.

Many outbreaks involving Salmonella enterica in dehydrated coconut have been reported. Little is known about the survival of S. enterica in dehydrated coconut flakes at common retail or domestic storage conditions. This study evaluated the behavior of a S. enterica cocktail (S. Enteritidis PT4, S. Typhimurium PT4, S. Bredeney, S. Muenster and S. Agona) in conventionally and osmotically dehydrated coconut flakes under four storage regimes: 25 °C for 120 days, 25 °C for 30 days then 7 °C for 90 days, 7 °C for 30 days then 25 °C for 90 days, and 7 °C for 120 days. S. enterica membrane integrity (using with propidium iodide and bis-1,3-dibutylbarbutyric acid) and metabolic activity (using 5-cyano-2,3-ditolyl tetrazolium chloride) were assessed by flow cytometry analysis after dehydration and storage at 7 °C or 25 °C for 120 days. Lower S. enterica inactivation rates (kmax 0.02 to 0.04 1/days) were observed in conventionally dehydrated coconut flakes compared to osmotically dehydrated coconut flakes (kmax 0.16 to 0.20 1/days). Changes in storage temperature did not affect the behavior of S. enterica in conventionally or osmotically dehydrated coconut flakes. Results show that S. enterica inactivation in conventionally dehydrated coconut flakes could be described by log-linear with tail models. S. enterica inactivation in osmotically dehydrated coconut flakes could be described by log-linear with shoulder and tail models. Subpopulations of S. enterica cells with damaged membranes and without metabolic activity were larger in conventionally (32.1% and 90.9%, respectively) than osmotically dehydrated coconut (18.5% and 82.2%, respectively) flakes at the beginning of the storage. Subpopulations of S. enterica cells with damaged membrane decreased by 9.4-14.4%, while cells with membrane potential and intact membrane increased by 23.7 and 24.2% in conventionally dehydrated coconut flakes after 120 days of storage at 7 °C or 25 °C, respectively. Subpopulations of S. enterica with damaged membranes did not change significantly in osmotically dehydrated coconut flakes. Our findings suggest that S. enterica populations decline during storage occurs due in part to membrane integrity losses. These data can contribute to the development of risk management strategies for S. enterica in dehydrated coconut flakes.

Control of Salmonella in low-moisture foods: Enterococcus faecium NRRL B-2354 as a surrogate for thermal and non-thermal validation.

Salmonella has been implicated in multiple foodborne outbreaks and recalls associated with low water activity foods (LawF). To verify the effectiveness of a process against Salmonella in LawF, validation using a nonpathogenic surrogate strain is essential. Enterococcus faecium NRRL B-2354 strain has been used as a potential surrogate of Salmonella in different processing of LawF. However, the survival of Salmonella and E. faecium in LawF during food processing is a dynamic function of aw, food composition and structure, processing techniques, and other factors. This review assessed pertinent literature on the thermal and non-thermal inactivation of Salmonella and its presumable surrogate E. faecium in various LawF and provided an overview of its suitibility in different LawF. Overall, based on the D-values, survival/reduction, temperature/time to obtain 4 or 5-log reductions, most studies concluded that E. faecium is a suitable surrogate of Salmonella during LawF processing as its magnitude of resistance was slightly greater or equal (i.e., statistical similar) as compared to Salmonella. Studies also showed its unsuitability which either does not provide a proper margin of safety or being overly resistant and may compromise the quality and organoleptic properties of food. This review provides useful information and guidance for future validation studies of LawF.

Induction of the Viable-but-Nonculturable State in Salmonella Contaminating Dried Fruit.

Salmonella can become viable but nonculturable (VBNC) in response to environmental stressors, but the induction of the VBNC state in Salmonella contaminating ready-to-eat dried fruit is poorly characterized. Dried apples, strawberries, and raisins were mixed with a five-strain cocktail of Salmonella at 4% volume per weight of dried fruit at 109 CFU/g. The inoculated dried fruit were then dried in desiccators at 25°C until the water activity (aw) approximated that of the uninoculated dried fruit. However, Salmonella could not be recovered after drying, not even after enrichment, suggesting a population reduction of approximately 8 log CFU/g. To assess the potential impact of storage temperature on survival, dried apples were spot-inoculated with the Salmonella cocktail, dried under ambient atmosphere at 25°C, and stored at 4 and 25°C. Spot inoculation permitted recovery of Salmonella on dried apple after drying, with the population of Salmonella decreasing progressively on dried apples stored at 25°C until it was undetectable after about 46 days, even following enrichment. The population decline was noticeably slower at 4°C, with Salmonella being detected until 82 days. However, fluorescence microscopy and laser scanning confocal microscopy with the LIVE/DEAD BacLight bacterial viability system at time points at which no Salmonella could be recovered on growth media even following enrichment showed that a large proportion (56 to 85%) of the Salmonella cells on the dried fruit were viable. The data suggest that the unique combination of stressors in dried fruit can induce large numbers of VBNC cells of Salmonella. IMPORTANCE Salmonella is a leading foodborne pathogen globally causing numerous outbreaks of foodborne illnesses and remains the leading contributor to deaths attributed to foodborne disease in the United States and other industrialized nations. Therefore, efficient detection methods for Salmonella contaminating food are critical for public health and food safety. Culture-based microbiological methods are considered the gold standard for the detection and enumeration of Salmonella in food. Findings from this study suggest that unique stressors on dried fruit can induce the VBNC state in Salmonella, thus rendering it undetectable with culture-based methods even though the bacteria remain viable. Therefore, strong consideration should be given to using, in addition to culture-based methods, microscopic and molecular methods for the accurate detection of all viable and/or culturable cells of Salmonella contaminating dried fruit, as all of these cells have the potential to cause human illness.