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Antimicrobial blue light (aBL) in the 400-470 nm wavelength range has been reported to kill multiple bacteria. This study assessed its potential for mitigating an important foodborne pathogen, Listeria monocytogenes (Lm), focusing on surface decontamination. Three wavelengths were tested, with gallic acid as a photosensitizing agent (Ps), against dried cells obtained from bacterial suspensions, and biofilms on stainless-steel (SS) coupons. Following aBL exposure, standard microbiological analysis of inoculated coupons was conducted to measure viability. Statistical analysis of variance was performed. Confocal laser scanning microscopy was used to observe the biofilm structures. Within 16 h of exposure at 405 nm, viable Lm dried cells and biofilms were reduced by approx. 3 log CFU/cm 2 with doses of 2,672 J/cm 2 . Application of Ps resulted in an additional 1 log CFU/cm 2 at 668 J/cm 2 , but its effect was not consistent. The highest dose (960 J/cm 2 ) at 420 nm reduced viable counts on the biofilms by 1.9 log CFU/cm 2 . At 460 nm, after 800 J/cm 2 , biofilm counts were reduced by 1.6 log CFU/cm 2 . The effect of material composition on Lm viability was also investigated. Irradiation at 405 nm (668 J/cm 2 ) of cells dried on polystyrene resulted in one of the largest viability reductions (4.0 log CFU/cm 2 ), followed by high-density polyethylene (3.5 log CFU/cm 2 ). Increasing the dose to 4,008 J/cm 2 from 405 nm (24 h), improved its efficacy only on SS and polyvinyl chloride. Biofilm micrographs displayed a decrease in biofilm biomass due to the removal of biofilm portions from the surface and a shift from live to dead cells suggesting damage to biofilm cell membranes. These results suggest that aBL is a potential intervention to treat Lm contamination on typical material surfaces used in food production.IMPORTANCECurrent cleaning and sanitation programs are often not capable of controlling pathogen biofilms on equipment surfaces, which transmit the bacteria to ready-to-eat foods. The presence of native plant microbiota and organic matter can protect pathogenic bacteria by reducing the efficacy of sanitizers as well as promoting biofilm formation. Post-operation washing and sanitizing of produce contact surfaces might not be adequate in eliminating the presence of pathogens and commensal bacteria. The use of a dynamic and harmless light technology during downtime and close of operation could serve as a useful tool in preventing biofilm formation and persistence. Antimicrobial blue light (aBL) technology has been explored for hospital disinfection with very promising results, but its application to control foodborne pathogens remains relatively limited. The use of aBL could be a complementary strategy to inactivate surfaces in restaurant or supermarket deli settings., Competing Interests: The authors declare no conflict of interest.

This work aimed to determine the ability of Listeria innocua (L.i.) to colonize eight materials found in food-processing and packaging settings and to evaluate the viability of the sessile cells. We also selected four commonly used phytochemicals (trans-cinnamaldehyde, eugenol, citronellol, and terpineol) to examine and compare their efficacies against L.i. on each surface. Biofilms were also deciphered in chamber slides using confocal laser scanning microscopy to learn more about how phytochemicals affect L.i. The materials tested were silicone rubber (Si), polyurethane (PU), polypropylene (PP), polytetrafluoroethylene (PTFE), stainless steel 316 L (SS), copper (Cu), polyethylene terephthalate (PET), and borosilicate glass (GL). L.i. colonized Si and SS abundantly, followed by PU, PP, Cu, PET, GL, and PTFE surfaces. The live/dead status ranged from 65/35% for Si to 20/80% for Cu, and the estimates of cells unable to grow on Cu were the highest, reaching even 43%. Cu was also characterized by the highest degree of hydrophobicity (ΔG TOT = -81.5 mJ/m 2 ). Eventually, it was less prone to attachment, as we could not recover L.i. after treatments with control or phytochemical solutions. The PTFE surface demonstrated the least total cell densities and fewer live cells (31%) as compared to Si (65%) or SS (nearly 60%). It also scored high in hydrophobicity degree (ΔG TOT = -68.9 mJ/m 2 ) and efficacy of phytochemical treatments (on average, biofilms were reduced by 2.1 log 10 CFU/cm 2 ). Thus, the hydrophobicity of surface materials plays a role in cell viability, biofilm formation, and then biofilm control and could be the prevailing parameter when designing preventive measures and interventions. As for phytochemical comparison, trans-cinnamaldehyde displayed greater efficacies, with the highest reductions seen on PET and Si (4.6 and 4.0 log 10 CFU/cm 2 ). The biofilms in chamber slides exposed to trans-cinnamaldehyde revealed the disrupted organization to a greater extent than other molecules. This may help establish better interventions via proper phytochemical selection for incorporation in environment-friendly disinfection approaches., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

This project was undertaken to determine the kinetic parameters of thermal inactivation of Listeria monocytogenes on pecans, macadamia nuts, and sunflower seeds subjected to heat treatments simulating industry processes. Five strains were grown in nonselective medium, mixed, and resuspended before inoculating macadamia nuts, pecans, and sunflower seeds (6 to 9 Log CFU/g). Redried inoculated pecans and macadamia nuts were heated in an oven at a temperature range of 90 to 140°C. Unshelled sunflower seeds were heated in sunflower seed oil. The thermal inactivation was determined by measuring viable cell counts using standard microbiological methods. Average count data were fit to the log-linear model, and thermal-death kinetics were calculated. On pecans, the viable Listeria counts were reduced by 3 and 3.5 Log CFU/g after 40 min at 110°C and 8 min at 140°C, respectively. On macadamia nuts, the L. monocytogenes population was reduced by 5 Log CFU/g after 20 min at 120°C. Unshelled sunflower seeds were subjected to heat treatment via a hot-oil bath. On sunflower seeds, >7 Log CFU/g reductions were observed after 15 min at 120°C. The thermal resistance ( D value) for inactivation on pecans at 140°C was 3.1 min and on macadamia nuts at 120°C was 4.4 min. The inactivation of L. monocytogenes was influenced by the kind of nut or seed. These results suggest that L. monocytogenes has a relatively high thermal tolerance. The findings from this study will contribute to the assessment of the effectiveness of heat treatment for control of this pathogen on nuts and seeds. IMPORTANCE Listeria monocytogenes is a major concern for the food industry in ready-to-eat (RTE) foods. In recent years, large-scale recalls have occurred with contaminated sunflower seeds and macadamia nuts that triggered product withdrawals. These events stress the importance of understanding Listeria' s ability to survive heat treatments in these low-water activity foods. Nuts and seeds are subjected to a variety of thermal treatments typically referred as roasting. To date, no listeriosis outbreak has been linked to nuts and seeds, but the recent recognition that this pathogen can be detected in commercial products stresses the need for research on thermal treatments. The characterization of heat inactivation kinetics at temperatures typically used during roasting processes will be very beneficial for validation studies. This manuscript reports inactivation rates of L. monocytogenes strains inoculated onto macadamia nuts, sunflower seeds, and pecan halves subjected to temperatures between 90 and 140°C.

Queso fresco, is a Hispanic non-fermented cheese highly susceptible to contamination with L. monocytogenes. This research was aimed to determine the effect of GRAS antimicrobial ingredients to control L. monocytogenes. Antimicrobials included caprylic acid (CA), Nisaplin ® (N, 2.5% nisin), a mixture of sodium lactate and sodium diacetate (SL/SD), Lactococcus lactis sbp. lactis DPC 3147, monolaurin, and lactic acid (LA). Batches of queso fresco curds were inoculated with 10 4  CFU/g and stored at 4 °C for three weeks. During storage the count of L. monocytogenes reached 7 to 8 Log CFU/g in control samples. Most individual antimicrobial treatments resulted in less than 1 Log CFU/g reductions in final counts, with the exception of N (0.5 g/kg) and CA (2.9 g/kg) that caused more than 3 and 5 Log CFU/g differences with controls, respectively. Mixtures of ingredients were more effective in inhibiting L. monocytogenes growth, and treatments with N and CA consistently delivered 6 Log CFU/g less counts than controls. Supplementation of 12 g/kg LA to treatments with SL/SD (3%/0.22%) caused differences of more than 4 Log CFU/g in final Listeria populations. Samples treated with the binary mixtures of N and CA (0.5 and 0.7 g/kg, respectively) were evaluated in a consumer panel (n = 67). Panelists slightly preferred control and commercial over treated samples, but all samples were in average rated between "slightly liking" and "moderately liking." These experiments indicated that combined use of antimicrobial ingredients may be an effective way to control the population of Listeria monocytogenes in queso fresco., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Queso fresco (QF), a fresh Hispanic cheese has been linked to outbreaks and recalls caused by Listeria contamination. The use antimicrobial treatments may be a potential solution. The goal of this research was to test the addition of nisin (N), caprylic acid (CA) and trans-cinnamaldehyde (CN) as anti-listerial ingredients in QF. QF batches were inoculated with approx. 10(4) CFU/g of 5- or 6-strain mixtures of Listeria monocytogenes and treated with antimicrobials. Samples were stored at 4 °C for three weeks and Listeria counts were determined by plating on PALCAM agar. The impact on the QF's natural indicator microorganisms was also assessed during refrigerated storage. All N and CA combinations (≥0.4 g/kg each) were effective against L. monocytogenes and reduced the final counts by at least 3 log CFU/g after 20 days of storage compared to controls. The levels of most strain mixtures were reduced immediately after treatment and their numbers remained below 10(3) CFU/g during storage. CN (1.2 g/kg) was bacteriostatic against L. monocytogenes, but it did not reduce initial counts. The addition of CN to the combination of N and CA did not enhance their antimicrobial effect. Results indicated that combinations of N and CA could control L. monocytogenes in QF with little impact on the natural flora of the cheese, providing a solution to control post processing L. monocytogenes contamination of QF., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

The growth of Listeria monocytogenes inoculated on frankfurters at four inoculum levels (0.1, 0.04, 0.01, and 0.007 CFU/g) was examined at 4, 8, and 12 degrees C until the time L. monocytogenes populations reached a detectable limit of at least 2 CFU/g. A scaled-down assumption was made to simulate a 25-g sample from a 100-lb batch size in a factory setting by using a 0.55-g sample from a 1,000-g batch size in a laboratory. Samples of 0.55 g were enriched in PDX-LIB selective medium, and presumptive results were confirmed on modified Oxford agar. Based on the time to detect (TTD) from each inoculum level and at each temperature, a shelf life model was constructed to predict the detection or risk levels reached by L. monocytogenes on frankfurters. The TTD increased with reductions in inoculum size and storage temperature. At 4 degrees C the TTDs (+/- standard error) observed were 42.0 +/- 1.0, 43.5 +/- 0.5, 50.7 +/- 1.5, and 55.0 +/- 3.0 days when the inoculum sizes were 0.1, 0.04, 0.01, and 0.007 CFU/g, respectively. From the same corresponding inoculum sizes, the TTDs at 8 degrees C were 4.5 +/- 0.5, 6.5 +/- 0.5, 7.0 +/- 1.0, and 8.5 +/- 0.5 days. Significant differences (P < 0.05) between TTDs were observed only when the inoculum sizes differed by at least 2 log. On a shelf life plot of 1n (TTD) versus temperature, the Q10 (increase in TTD for a 10 degrees C increase in temperature) values ranged from 24.5 to 44.7 and with no significant influence from the inoculum densities. When the observed TTDs were compared with the expected detection times based on the data obtained from a study with an inoculum size of 10 to 20 CFU/g, significant deviations were noted at lower inoculum levels. These results can be valuable in designing a safety-based shelf life model for frankfurters and in performing quantitative risk assessment of listeriosis at low and practical contamination levels.

This research was conducted to study the growth of Listeria monocytogenes inoculated on frankfurters stored at different conditions as a basis for a safety-based consume by shelf life date label. Three L. monocytogenes strains were separately inoculated at 10 to 20 CFU/cm2 onto frankfurters that were previously formulated with or without high pressure and with or without added 2% potassium lactate (PL) and 0.2% sodium diacetate (SD). Inoculated frankfurters were air or vacuum packaged; stored at 4, 8, or 12 degrees C; and L. monocytogenes and psychrotrophic plate counts were determined for 90, 60, and 45 days, respectively, or until the stationary phase was reached. The data (log CFU per square centimeter versus time) were fitted using the Baranyi-Roberts model to determine maximum growth rates and lag-phase time. The maximum growth rates and the lag time under each growth condition were used to calculate the time to reach 100-fold the initial Listeria population. In frankfurters lacking PL and SD, the count of all strains increased by 2 log after 18 to 50 days at 4 degrees C and 4 to 13 days at 8 degrees C. The growth was inhibited at 4 and 8 degrees C in frankfurters containing PL and SD, but one ribotype was capable of growing, with the time to reach 100-fold the initial Listeria population ranging from 19 to 35 days at 12 degrees C. In most cases, the time to reach 100-fold the initial Listeria population of L. monocytogenes was significantly longer in vacuum-packaged frankfurters as compared with air-packaged samples. Inclusion of PL and SD also inhibited the growth of psychrotrophs, but at all temperatures the psychrotrophic plate counts were greater than 4 log CFU/cm2 at the end of the experiments. These results indicated that despite the use of antimicrobials, certain L. monocytogenes strains could be capable of growing under storage-abuse conditions. Growth kinetics data could be useful for establishing a shelf life date label protocol under different handling scenarios.

The growth variability of three Listeria monocytogenes ribotypes in ready-to-eat (RTE) sliced uncured turkey breast and cured ham was studied under storage conditions that RTE foods are likely to encounter. Three product treatments studied were: (1) a control; (2) a formulation subjected to high pressure processing to reduce initial microbial load (HPP); (3) a formulation containing 2.0% potassium lactate and 0.2% sodium diacetate (PL/SD). After separate inoculation with individual L. monocytogenes ribotypes and packaging each treatment under air and vacuum, the packages were stored at 4, 8, or 12 degrees C and the counts of L. monocytogenes and psychrotrophic bacteria (PPC) were determined for several weeks. The Baranyi model was used to estimate lag times and growth rates. Significant effect of strain difference was noted in both sliced products (P<0.05). In the absence of antimicrobials (HPP and control), the growth rate (GR) of L. monocytogenes strains increment from 4 to 8 degrees C and from 8 to 12 degrees C was approximately 10 and 2 fold, respectively. The addition of PL/SD was effective in restricting the growth of L. monocytogenes and PPC at 4 degrees C, but at 8 and 12 degrees C significant growth was observed (more than 100-fold increase) (P<0.05). In PL/SD samples, vacuum packaging slowed down the onset and the rate of growth of L. monocytogenes at 12 degrees C in sliced ham and at 8 and 12 degrees C in sliced turkey breast. Generally, the time to increase by 2-logs was greater in control samples than as observed in HPP-treated samples. When antimicrobials were present, the current results showed that L. monocytogenes was able to grow more than 100-fold within the typical quality-based shelf life of 60 to 90 days at 8 and 12 degrees C. The findings of this study should be useful in setting the duration of a safety-based shelf life for RTE sliced meat and poultry foods.

This study compared the performance of four primary mathematical models to study the growth kinetics of Listeria monocytogenes ribotypes grown at low temperature so as to identify the best predictive model. The parameters of the best-fitting model were used to select the fastest growing strains with the shortest lag time and greatest growth rate. Nineteen food, human and animal L. monocytogenes isolates with distinct ribotype were grown at 4, 8, and 12 degrees C in tryptic soy broth and slurries prepared from cooked uncured sliced turkey breasts (with or without potassium lactate and sodium diacetate, PL/SD) and cooked cured frankfurters (with or without PL/SD). Separate regressions were performed on semi-logarithm growth curves to fit linear (based on Monod) and non-linear (Gompertz, Baranyi-Roberts, and Logistic) equations and performance of each model was evaluated using an F-test. No significant differences were found in the performance of linear and non-linear models, but the Baranyi model had the best fit for most growth curves. The maximum growth rate (MGR) of Listeria strains increased with the temperature. Similarly MGR was found significantly greater when no antimicrobials were present in the formulation of turkey or frankfurter products. The variability in lag times and MGRs in all media as determined by the Baranyi model was not consistent among strains. No single strain consistently had the fastest growth (shortest lag time, fastest MGR, or shortest time to increase 100-fold), but nine strains were identified as fastest growing strains under most growth conditions. The lack of association between serotype and fastest strain was also observed in the slurry media study. The fastest growing strains resulting from this study can be recommended for future use in L. monocytogenes challenge studies in delicatessen meat and poultry food matrices, so as to develop conservative pathogen growth predictions.

In the current study, temperature dependent growth of Listeria monocytogenes strain H7776, with an initial inoculum level of approximately 0.1 CFU/mL or g, was modeled based on "time to detect" (TTD) using the Arrhenius equation. The activation energies (E(a)) obtained from specific growth rate and lag phase duration in tryptic soy broth (TSB) and frankfurters were compared. At 4 degrees C, the TTD for L. monocytogenes was 217 h in TSB and 48 h in portions of frankfurters. The TTD decreased as temperature increased from 4 to 36 degrees C in both culture media and frankfurters, and this relationship was modeled using the Arrhenius equation. Based on this model, the E(a) values for the TTD in TSB and frankfurters averaged 22.7 and 18.7 kcal/mol, respectively, and were not significantly different (p<0.05). Linear regression was performed on the exponential part of the growth curve to evaluate specific growth rate constants at each temperature using the Monod model. The E(a) values were also calculated based on the specific growth rate and the lag phase of L. monocytogenes in TSB and frankfurters incubated in the same range of temperatures. The average E(a) values for the specific growth rates and the lag phase durations in TSB cultures were 19 and 21 kcal/mol, respectively. In frankfurters, the average E(a) values were slightly greater for both specific growth rate and lag phase duration (29 and 35 kcal/mol, respectively), but these values were not significantly different from the E(a) calculated for the TTD in each medium. These results indicate that the TTD concept can be used to develop and validate safety-based shelf life models.

In nature, Listeria may interact competitively and cooperatively with other organisms, resulting in unique spatial organization and functions for cells within the community. This study was undertaken to characterize the biofilm architecture of binary biofilms of Listeria monocytogenes and Lactobacillus species and to assess their effect on the survival of Listeria during exposure to hypochlorite. Three L. monocytogenes strains, ATCC 19115 (Lm5), ATCC 19117 (Lm7), and Coleslaw (LmC), were selected and combined individually with three Lactobacillus strains: L. fermentum (Lf), L. bavaricus (Lb), and L. plantarum (Lp). In binary Lm-Lp biofilms, the Lm cell counts were similar to single-species biofilms (8.5 log CFU/well), and the Lp cell numbers declined by 1.0 log CFU/well. In the presence of Lb, the Lm cell counts were reduced by 1.5 log CFU/well (p < 0.05), whereas the Lf cell counts increased at least by 3.5 log CFU/well. Confocal laser scanning microscopy (CLSM) determined that interspecies interactions significantly affected the spatial organization of three binary biofilms. Biofilm surface-to-volume ratio increased from 0.8 μm2/μm3 for Lm5 in the monoculture to 2.1 μm2/μm3 for Lm5-Lp in the dual-species model (p < 0.05), and was characterized by a thicker structure with a largely increased surface area. Biofilm roughness increased from 0.2 for Lm7 to 1.0 for Lm7-Lb biofilms (p < 0.05), which appeared as interspecific segregation. Biofilm thickness increased from 34.2 μm for LmC to 46.3 μm for LmC–Lf (p < 0.05), which produced flat and compact structures that covered the entire surface available. The biomass of the extracellular matrix was higher in the case of some binary biofilms (p < 0.05); however, this effect was dependent upon the species pair. When treated with hypochlorite, Lm5 in binary biofilms had an approximately 1.5 log CFU/well greater survival than individually. The unique spatial organization and greater protein production may explain the protective effect of Lp after hypochlorite exposure. [ABSTRACT FROM AUTHOR]

Abstract: Electrochemically activated water (ECAW), also known as electrolyzed water, and ozonized water are typically effective in inactivating bacteria, but their generation typically uses high current and voltage. A few simpler antimicrobial technologies that are also based on the application of a mild electrical current have been recently marketed to food retail and service customers claiming to have sanitizing properties for controlling bacteria. The objective of this study was to determine the sanitizing effect of some of these commercial technologies on Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella enterica and compare them with sterile water, generated ECAW generated with a pilot size electrolyzing unit, and salt solutions sprayed using commercial device sprays. A concentration of 100 mg/L ECAW had sanitizing effects of at least 5 log CFU/mL reductions on liquid culture and more than 4 log CFU/coupon reductions for E. coli O157:H7, L. monocytogenes and Salmonella dried on stainless steel surface, respectively. No bacterial cells were detected by direct plate counting post-ECAW treatment. In contrast, the treatment of liquid cultures with any of the commercial technologies tested resulted in non-significant bacterial cell reductions greater than 0.5 log CFU/mL. Similarly, when cells had been dried on metal surfaces and treated with any of the water generated with those technologies, no reductions were observed. When the manufacturer's instructions were followed, the reduction of cells on surface was largely due to the physical removal by cloth-wiping after water fraction application. These results indicate that treatment with any of these portable technologies had no noticeable antimicrobial activity. These results would be helpful for guiding consumers when choosing a right sanitization to ensure food safety. [Copyright &y& Elsevier]

Bacteriophages are considered a potential antimicrobial food intervention due to their ability to inactivate target bacteria in different environments. This study was undertaken to isolate, characterize, and identify a mixture of phages that could be used for control of Listeria monocytogenes . A total of 37 phages were isolated from raw sewage sludge using L. monocytogenes serotypes 4b, 1/2a and 1/2b strains. The host range of these phages was determined by spot-testing. Ten lytic phages were selected for further evaluation with efficiency of plating (EOP) analysis and characterization by pulsed field gel electrophoresis (PFGE). According to the spot tests and EOP results, the isolated phages against L. monocytogenes 4b strains were found to be capable of lysing a wider range of ribotypes. Also, three phages specific for 1/2a and 1/2b serotypes had similar lysis capabilities over a range of target Listeria with high EOP values between 0.5 and 1.0. PFGE analysis identified 5 distinct restriction profiles of the 10 selected phages whose DNA was digested with either Eco RI or Hin DIII. Transmission electron microscopy (TEM) pictures revealed that five selected phages belonged to the Myoviridae and one phage to the Podoviridae family. Based on screening results, a phage cocktail (LP1) comprised of six lytic phages was used to inactivate a mixed culture of five L. monocytogenes strains. LP1 was able to lyse bacterial cells within 30–60 min at multiplicity of infection values (MOI, PFU/CFU) of 1000 and maintained the target cells under detection levels for 24 to 48 h at 4, 10, and 30 °C. These results suggested the LP1 phage collection contained effective and strongly lytic phages that may serve as antibacterial preparation to control L. monocytogenes of different serotypes. [ABSTRACT FROM AUTHOR]