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.

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.

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.

The outbreaks linked to foodborne illnesses in low-moisture foods are frequently reported due to the occurrence of pathogenic microorganisms such as Salmonella Spp. Bacillus cereus, Clostridium spp., Cronobacter sakazakii, Escherichia coli, and Staphylococcus aureus. The ability of the pathogens to withstand the dry conditions and to develop resistance to heat is regarded as the major concern for the food industry dealing with low-moisture foods. In this regard, the present review is aimed to discuss the importance and the use of novel thermal and nonthermal technologies such as radiofrequency, steam pasteurization, plasma, and gaseous technologies for decontamination of foodborne pathogens in low-moisture foods and their microbial inactivation mechanisms. The review also summarizes the various sources of contamination and the factors influencing the survival and thermal resistance of pathogenic microorganisms in low-moisture foods. The literature survey indicated that the nonthermal techniques such as CO2 , high-pressure processing, and so on, may not offer effective microbial inactivation in low-moisture foods due to their insufficient moisture content. On the other hand, gases can penetrate deep inside the commodities and pores due to their higher diffusion properties and are regarded to have an advantage over thermal and other nonthermal processes. Further research is required to evaluate newer intervention strategies and combination treatments to enhance the microbial inactivation in low-moisture foods without significantly altering their organoleptic and nutritional quality.

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.

Enteric viruses, such as human norovirus (NoV) and hepatitis A virus (HAV), are the major causes of foodborne illnesses worldwide. These viruses have low infectious dose, and may remain infectious for weeks in the environment and food. Limited information is available regarding viral survival and transmission in low-moisture foods (LMF). LMFs are generally considered as ready-to-eat products, which undergo no or minimal pathogen reduction steps. However, numerous foodborne viral outbreaks associated with LMFs have been reported in recent years. The objective of this study was to examine the survival of foodborne viruses in LMFs during 4-week storage at ambient temperature and to evaluate the efficacy of advanced oxidative process (AOP) treatment in the inactivation of these viruses. For this purpose, select LMFs such as pistachios, chocolate, and cereal were inoculated with HAV and the norovirus surrogates, murine norovirus (MNV) and feline calicivirus (FCV), then viral survival on these food matrices was measured over a four-week incubation at ambient temperature, by both plaque assay and droplet-digital RT-PCR (ddRT-PCR) using the modified ISO-15216 method as well as the magnetic bead assay for viral recovery. We observed an approximately 0.5 log reduction in viral genome copies, and 1 log reduction in viral infectivity for all three tested viruses following storage of select inoculated LMFs for 4 weeks. Therefore, the present study shows that the examined foodborne viruses can persist for a long time in LMFs. Next, we examined the inactivation efficacy of AOP treatment, which combines UV-C, ozone, and hydrogen peroxide vapor, and observed that while approximately 100% (4 log) inactivation can be achieved for FCV, and MNV in chocolate, the inactivation efficiency diminishes to approximately 90% (1 log) in pistachios and 70% (< 1 log) in cereal. AOP treatment could therefore be a good candidate for risk reduction of foodborne viruses from certain LMFs depending on the food matrix and surface of treatment.

The contamination risks of microorganisms and mycotoxins in low-moisture foods have heightened public concern. Developing novel decontamination technologies to improve the safety of low-moisture foods is of great interest in both economics and public health. This review summarizes the working principles and applications of novel thermal decontamination technologies such as superheated steam, infrared, microwave, and radio-frequency heating as well as extrusion cooking. These methods of decontamination can effectively reduce the microbial load on products andmoderately destruct the mycotoxins. Meanwhile, several integrated technologies have been developed that take advantage of synergistic effects to achieve the maximum destruction of contaminants and minimize the deterioration of products.

Salmonella spp. are resilient bacterial pathogens in low-moisture foods. There has been a general lack of understanding of critical factors contributing to the enhanced thermal tolerance of Salmonella spp. in dry environments. In this study, we hypothesized that the moisture content (XW ) of bacterial cells is a critical intrinsic factor influencing the resistance of Salmonella spp. to thermal inactivation. We selected Salmonella enterica serotype Enteritidis PT 30 to test this hypothesis. We first produced viable freeze-dried S. Enteritidis PT 30, conditioned the bacterial cells to different XW s (7.7, 9.2, 12.4, and 15.7 g water/100 g dry solids), and determined the thermal inactivation kinetics of those cells at 80°C. The results show that the D-value (the time required to achieve a 1-log reduction) decreased exponentially with increasing XW We further measured the water activities (aw) of the freeze-dried S. Enteritidis PT 30 as influenced by temperatures between 20 and 80°C. By using those data, we estimated the XW of S. Enteritidis PT 30 from the published papers that related the D-values of the same bacterial strain at 80°C with the aw of five different food and silicon dioxide matrices. We discovered that the logarithmic D-values of S. Enteritidis PT 30 in all those matrices also decreased linearly with increasing XW of the bacterial cells. The findings suggest that the amount of moisture in S. Enteritidis PT 30 is a determining factor of its ability to resist thermal inactivation. Our results may help future research into fundamental mechanisms for thermal inactivation of bacterial pathogens in dry environments.IMPORTANCE This study established a logarithmic relationship between the thermal death time (D-value) of S. Enteritidis PT 30 and the moisture content (XW ) of the bacterial cells by conducting thermal inactivation tests on freeze-dried S Enteritidis PT 30. We further verified this relationship using literature data for S. Enteritidis PT 30 in five low-moisture matrices. The findings suggest that the XW of S. Enteritidis PT 30, which is rapidly adjusted by microenvironmental aw, or relative humidity, during heat treatments, is the key intrinsic factor determining the thermal resistance of the bacterium. The quantitative relationships reported in this study may help guide future designs of industrial thermal processes for the control of S. Enteritidis PT 30 or other Salmonella strains in low-moisture foods. Our findings highlight a need for further fundamental investigation into the role of water in protein denaturation and the accumulation of compatible solutes during thermal inactivation of bacterial pathogens in dry environments.

Salmonella enterica is the leading foodborne pathogen associated with outbreaks involving low-moisture foods (LMFs). However, the genes involved in Salmonella\'s long-term survival on LMFs remain poorly characterized. In this study, in-shell pistachios were inoculated with Tn5-based mutant libraries of S. Enteritidis P125109, S. Typhimurium 14028s, and S. Newport C4.2 at approximate 108 CFU/g and stored at 25°C. Transposon sequencing analysis (Tn-seq) was then employed to determine the relative abundance of each Tn5 insertion site immediately after inoculation (T0), after drying (T1), and at 120 days (T120). In S. Enteritidis, S. Typhimurium, and S. Newport mutant libraries, the relative abundance of 51, 80, and 101 Tn5 insertion sites, respectively, was significantly lower at T1 compared to T0, while in libraries of S. Enteritidis and S. Typhimurium the relative abundance of 42 and 68 Tn5 insertion sites, respectively, was significantly lower at T120 compared to T1. Tn5 insertion sites with reduced relative abundance in this competition assay were localized in DNA repair, lipopolysaccharide biosynthesis and stringent response genes. Twelve genes among those under strong negative selection in the competition assay were selected for further study. Whole gene deletion mutants in ten of these genes, sspA, barA, uvrB, damX, rfbD, uvrY, lrhA, yifE, rbsR, and ompR, were impaired for individual survival on pistachios. The findings highlight the value of combined mutagenesis and sequencing to identify novel genes important for the survival of Salmonella in low-moisture foods.

Salmonella spp. is one of the top foodborne pathogens associated with low-moisture foods and they exhibit significant resistance to conventional thermal treatments. UV light pulses emitted from light emitting diode (LED) has shown antimicrobial potential in high-moisture foods and water. However, limited information is available about the antimicrobial potential of UV light with different wavelengths, including 395 nm in low-moisture foods. The objectives of this study were to investigate the antimicrobial potential of 395 nm pulsed LED light in wheat flour and the resulting quality changes. This study demonstrated a maximum 2.91 log reduction of Salmonella cocktail in wheat flour treated with 395 nm pulsed LED for 60 min in a semi-closed system. Oxidation occurred in wheat flour after 30 and 60 min exposure to the 395 nm LED, which subsequently led to bleaching, and polymerization of gluten components through disulphide linkage. The water holding capacity of gluten was reduced by oxidation, and the contents of secondary structures were altered significantly after pulsed LED treatment, but the rheological properties were not deteriorated. The disulfide bond formation naturally happens during dough formation and the oxidation triggered by pulsed LED treatment may play a role on accelerating this process. The 395 nm pulsed LED treatment could be a promising decontamination technology for wheat flour with an additional benefit of bleaching of the flour without chemicals. INDUSTRIAL RELEVANCE: A number of foodborne outbreaks and recalls have been related to low-moisture foods in these decades and recently several outbreaks were reported due to the occurrence of Salmonella in wheat flour. However, it is difficult to solve this problem through conventional thermal approaches because of the increased thermal resistance of Salmonella at low water activity environment. The emerging LED light source can produce light with monochromatic wavelengths without the use of mercury vapor lamps. It also has high durability, low heat generation, and is relatively easy to be adapted in an existing production line. Therefore, there is a great potential of using certain UV wavelengths emitted from LED to disinfect the low-moisture foods in food industries. To the best of our knowledge, no research was conducted on decontamination of wheat flour by using LEDs and only limited studies are available on the influence of pulsed LED treatment on food quality. The aim of this study was to explore the possibility of using 395 nm pulsed LED treatment as a novel tool for decontamination of Salmonella in a low-moisture food product (wheat flour) with industrial feasibility, and investigate the influence of the pulsed LED treatment on quality changes in the product.