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Wood is reportedly more difficult to maintain in hygienic condition versus other food contact materials, yet its use in produce packing and retail warrants efforts to reduce the risk of microbial pathogen contamination and attachment. This study characterized antifouling capabilities of fluorinated silanes applied to wood used in fresh edible produce handling to render the wood superhydrophobic and less supportive of bacterial pathogen attachment. Pine and oak cubic coupon surfaces were treated with 1% (w/w) silane or left untreated. Treated and untreated coupons were inoculated with Salmonella enterica or Listeria monocytogenes and held to facilitate pathogen attachment for 1, 4, or 8 h. Silane treatment of wood produced significant reductions in the proportions of strongly attaching cells for both pathogens versus loosely attaching cells (P < 0.01). Salmonella attachment demonstrated a dependency on wood treatment; silane-treated wood supported a lower fraction of strongly adhering cells (1.87 ± 1.24 log CFU/cm2) versus untreated wood (3.72 ± 0.67 log CFU/cm2). L. monocytogenes demonstrated significant declines in strongly attaching cells during extended exposure to silane-treated wood, from 7.59 ± 0.14 to 5.27 ± 0.68 log CFU/cm2 over 8 h post-inoculation. Microscopic analysis demonstrated silane treatment increased the surface roughness of both woods, leading to superhydrophobic conditions on wood surfaces, consequently decreasing strong attachment of pathogenic bacteria. • Fluorinated silane application rendered pine and oak wood superhydrophobic. • Silane-treatment produced significantly less strong adherence of pathogens (P < 0.01). • Silanes produced increased nano-scale roughness of treated versus non-treated wood. • Pathogen counts differed by wood treatment and/or time of post-inoculation contact. [ABSTRACT FROM AUTHOR]

The plasmid of emerging S. Infantis (pESI) or pESI-like plasmid in Salmonella enterica Infantis are consistently reported in poultry and humans worldwide. However, there has been limited research on these plasmids of S. Infantis isolated from eggs. Therefore, this study aimed to analyze the prevalence and characteristics of S. Infantis carrying the pESI-like plasmid from eggs in egg grading and packing plants. In this study, the pESI-like plasmid was only detected in 18 (78.3%) of 23 S. Infantis isolates, and it was absent in the other 9 Salmonella serovars. In particular, S. Infantis isolates carrying the pESI-like plasmid showed the significantly higher resistance to β-lactams, phenicols, cephams, aminoglycosides, quinolones, sulfonamides, and tetracyclines than Salmonella isolates without the pESI-like plasmid (p < 0.05). Moreover, all S. Infantis isolates carrying the pESI-like plasmid were identified as extended-spectrum β-lactamase (ESBL) producer, harboring the bla CTX-M-65 and bla TEM-1 genes, and carried non-β-lactamase resistance genes (ant(3′′)-Ia , aph(4)-Ia , aac(3)-IVa , aph(3′)-Ic , sul1 , tetA , dfrA14 , and floR) against five antimicrobial classes. However, all isolates without the pESI-like plasmid only carried the bla TEM-1 gene among the β-lactamase genes, and either had no non-β-lactamase resistance genes or harbored non-β-lactamase resistance genes against one or two antimicrobial classes. Furthermore, all S. Infantis isolates carrying the pESI-like plasmid carried class 1 and 2 integrons and the aadA1 gene cassette, but none of the other isolates without the pESI-like plasmid harbored integrons. In particular, D87Y substitution in the gyrA gene and IncP replicon type were observed in all the S. Infantis isolates carrying the pESI-like plasmid but not in the S. Infantis isolates without the pESI-like plasmid. The distribution of pulsotypes between pESI-positive and pESI-negative S. Infantis isolates was clearly distinguished, but all S. Infantis isolates were classified as sequence type 32, regardless of whether they carried the pESI-like plasmid. This study is the first to report the characteristics of S. Infantis carrying the pESI-like plasmid isolated from eggs and can provide valuable information for formulating strategies to control the spread of Salmonella in the egg industry worldwide. • The majority of S. Infantis isolated from eggs carried the pESI-like plasmid. • The pESI-like plasmid was associated with MDR and AMR genes. • S. Infantis carrying the pESI-like plasmid showed the same pulsotype. [ABSTRACT FROM AUTHOR]

This study assessed the fate of a Salmonella enterica cocktail (S. Typhimurium, S. Enteritidis, S. Newport, S. Agona and S. Anatum; initial counts 3.5 log CFU/g) in minimally processed sliced chard, broccoli and red cabbage at 16 conditions of different temperature (7, 14, 21 and 37 °C) and relative humidity (RH; 15, 35, 65 and 95%) over six days (144 h). Linear regression was used to estimate the rate change of Salmonella in cut vegetables as a function of temperature and relative humidity (RH). R2 value of 0.85, 0.87, and 0.78 were observed for the rates of change in chard, broccoli, and red cabbage, respectively. The interaction between temperature and RH was significant in all sliced vegetables. Higher temperatures and RH values favored Salmonella growth. As temperature or RH decreased, the rate of S. enterica change varied by vegetable. The models developed here can improve risk management of Salmonella in fresh cut vegetables. • Temperature and relative humidity (RH) affected S. enterica behavior in cut produce. • Salmonella survival rates differed in chard, broccoli and red cabbage. • Salmonella growth rate increased at higher temperatures and RH values. • Lower temperatures and RH contributed to S. enterica population decline. • Regression models predicted change in Salmonella populations in cut vegetables. [ABSTRACT FROM AUTHOR]

Isolation of Salmonella from enrichment cultures of food or environmental samples is a complicated process. Numerous factors including fitness in various selective enrichment media, relative starting concentrations in pre-enrichment, and competition among multi-serovar populations and associated natural microflora, come together to determine which serovars are identified from a given sample. A recently developed approach for assessing the relative abundance (RA) of multi-serovar Salmonella populations (CRISPR-SeroSeq or Deep Serotyping, DST) is providing new insight into how these factors impact the serovars observed, especially when different selective enrichment methods are used to identify Salmonella from a primary enrichment sample. To illustrate this, we examined Salmonella -positive poultry pre-enrichment samples through the selective enrichment process in Tetrathionate (TT) and Rappaport Vassiliadis (RVS) broths and assessed recovery of serovars with each medium. We observed the RA of serovars detected post selective enrichment varied depending on the medium used, initial concentration, and competitive fitness factors, all which could result in minority serovars in pre-enrichment becoming dominant serovars post selective enrichment. The data presented provide a greater understanding of culture biases and lays the groundwork for investigations into robust enrichment and plating media combinations for detecting Salmonella serovars of greater concern for human health. • Different Salmonella serovars are enriched from selective enrichments in RVS and TT. • Choice of plating media influences serovars recovered from selective enrichments. • Experiments revealed starting serovar relative concentrations influence isolation. • Robust methods are vital for reliably isolating Salmonella serovars of concern. [ABSTRACT FROM AUTHOR]

Salmonella is a major foodborne pathogen and the cause of significant morbidity and mortality via consumption of contaminated meat and meat-products. The prevalence of Salmonella in ducks and wild geese in China are poorly characterized and these sources represent a potential pool that could be transferred to farm-reared fowl. In this study, we isolated 335 (18.3%) Salmonella from 1830 samples and identified 24 serotypes and most prevalent were Salmonella Indiana, Salmonella Kentucky and Salmonella Typhimurium. Whole genome sequencing revealed the presence of the dominant sequence types ST17, ST198 and ST19 for these three serotypes, respectively. In addition, these isolates were most likely clonally spread across different regions while S. Kentucky also crossed the species barrier. The majority of the Salmonella isolates possessed β-lactam and fluoroquinolone resistance and these were consistent with antibiotic resistance gene profiles. We also identified 8 plasmid replicon types and all isolates possessed virulence genes and the numbers were greatest for S. Enteritidis and S. Typhimurium isolates. This study provides novel insights concerning the epidemiology of Salmonella in ducks and wild geese and provides basic data for public health screening and management. • 335 Salmonella isolates were recovered from 1830 ducks and wild geese samples across various regions in China. • Indiana, Kentucky, and Typhimurium emerged as the dominant ones among the identified serotypes. • ST17, ST198, and ST19 were identified as the predominant sequence types in three serotypes of Salmonella. • The transmission of Salmonella occurred through clonal expansion within specific geographic areas. • The trend of high resistance rates to first-line antibiotics is worrying. [ABSTRACT FROM AUTHOR]

This study determined the susceptibility to sanitizers and biofilm-forming ability on stainless steel of 43 Salmonella enterica and Listeria monocytogenes strains. Besides, the biofilm resistance to sanitizers of four bacterial pathogen strains was evaluated. Four sanitizers commonly used in the food industry were tested: peracetic acid (PAA), chlorine dioxide (ClO 2), sodium hypochlorite (SH), and quaternary ammonium compound (QAC). The susceptibility to sanitizers varied widely among the strains of both pathogens. On the other hand, the number of biofilm-associated cells on the stainless-steel surface was >5 log CFU/cm2 for all of them. Only one Salmonella strain and two L. monocytogenes strains stood out as the least biofilm-forming. The resistance of biofilms to sanitizers also varied among strains of each pathogen. Biofilms of L. monocytogenes were more susceptible to the disinfection process with ClO 2 and QAC than those of Salmonella. However, no correlation was observed between the ability to form denser biofilm and increased sanitizer resistance. In general, chlorine compounds were more effective than other sanitizers in inactivating planktonic cells and biofilms. • Biofilm formation and tolerance to sanitizers of 43 strains were assessed • L. monocytogenes strains were more susceptible to sanitizers than Salmonella • ClO 2 was more effective than QAC against Salmonella and L. monocytogenes strains • Biofilms of both pathogens showed high variability for resistance to sanitizers • The ability to form denser biofilm did not correlate with resistance to sanitizers [ABSTRACT FROM AUTHOR]

Superheated steam (SHS) is a powerful technology used to reduce bacteria on food surfaces while causing less damage to the underlying sublayer of food compared to conventional heating treatments. In this study, a semi-continuous SHS system was developed to inactivate foodborne pathogens within 1 s (Escherichia coli O157:H7, Salmonella enterica , and Listeria monocytogenes) on radish seed surfaces and to enhance the seeds' salinity tolerance, which is vital for adapting to arid and semi-arid regions. The temperature of the SHS was set to 200 °C and 300 °C, with flow rates of 5 m/s and 7 m/s, and treatments were cycled either once or three times. As a result, increased temperature (200 °C–300 °C) and number of treatments (1 time to 3 times) led to a significantly larger microbial reduction on the surface of radish seeds. E. coli O157:H7, S. enterica , and L. monocytogenes were reduced by 4.42, 4.73, and 3.95 log CFU/g (P < 0.05), respectively, after three SHS treatments at 300 °C and 7 m/s. However, due to the ongoing potential for recovery of residual microorganisms, further research involving combinations is essential to enhance the microbicidal effect. Water imbibition showed significantly higher values in the SHS-treated group up to 30 min, indicating faster germination rates in the SHS-treated group (71.3–81.3%) compared to the control group (52.7%) on the second day, indicating a significant enhancement in germination rate. In addition, the salinity resistance of the radish seeds increased after SHS treatment. When moisturized with 0.5% NaCl solution, more radish seeds germinated after treatment with SHS (40%) than controls (22.7%) (P < 0.05). The results of this study, the first to apply semi-continuous SHS to seeds, are expected to serve as a cornerstone for future pilot-scale investigations aiming to implement the system within the seed industry. • Semi-continuous SHS at 300 °C ensures biological safety by achieving over 3 log-reduction of pathogens on radish seeds. • Microbicidal effects of SHS are enhanced by higher fluid velocity and repeated treatment cycles. • Semi-continuous SHS system breaks seed dormancy, leading to improved water uptake, germination, and salinity tolerance. • The increased salinity tolerance will be beneficial when sprouting seeds using irrigation water, especially in arid regions. [ABSTRACT FROM AUTHOR]

This study assessed the adhesion and formation of biofilm by five Salmonella enterica strains (S. Enteritidis 132, S. Infantis 176, S. Typhimurium 177, S. Heidelberg 281 and S. Corvallis 297) on polypropylene (PP) and glass (G) surfaces as affected by pH (4–7), NaCl concentration (0–10% w/v) and temperature (8–35 °C). Sessile counts <3 log CFU/cm2 were considered lack of adhesion (category 1), while counts ≥ 3 and < 5 log CFU/cm2 corresponded to adhesion (category 2) and counts ≥ 5 log CFU/cm2 corresponded biofilm formation (category 3). The obtained results categorized in these three responses were used to develop ordinal regression models to predict the probability of biofilm stages on PP- and G-surfaces. The experimental outcomes for lack of adhesion were >90% on PP- and G-surfaces. Generally, adhesion outcomes corresponded to approximately 36% of the total, whereas biofilm outcomes were close to 65% in both PP- and G-surfaces. The biofilm stages varied among the strains studied and with the material surface under the same experimental conditions. According to the generated ordinal models, the probability of adhesion and biofilm formation on PP-surface by the five S. enterica strains tested decreased at pH 4 or 5 in NaCl concentrations >4% and at a temperature <20 °C. On G-surface, the probability of adhesion increased pH 6 or 7, in the absence of NaCl and temperatures <20 °C, while, the probability of biofilm formation increased in the same pH, NaCl concentration up to 4% and temperatures ≥20 °C. This is the first study assessing the biofilm formation through categorical, ordinal responses and it shows that ordinal regression models can be useful to predict biofilm stages of S. enterica as a function of pH, NaCl, and temperature or their interactions. • Biofilm formation on glass (G) or polypropylene (PP) varied among Salmonella serovars. • Salmonella serovars were able to adhere and form biofilm at pH 4 or 5 up to 6% NaCl. • Changes in pH, NaCl concentration or temperature (T) affect the biofilm stages. • Probability of adhesion on G increased at pH ≥ 6, absence of NaCl and T < 20 °C. • Probability of biofilm on PP decreased at pH 4 or 5, NaCl > 4% and T < 20 °C. [ABSTRACT FROM AUTHOR]

Fresh vegetables are important components of an everyday balanced diet making ready to-eat-salads (RTE) a commodity widely consumed. However, in the past few years these products have been linked with outbreaks of salmonellosis and listeriosis; thus the continuous investigation of their safety is an essential requirement. A total of 216 samples of ready-to-eat salads from the Cypriot market were analysed to determine the microbiological quality and safety, along with physicochemical attributes of the salads and identify possible correlations between them. The samples were randomly collected from four retail outlets and correspond to five different salad producing companies. Furthermore, the effects of season, salad producer and type of salad and/or their interactions with the tested parameters were investigated. The results revealed that the higher microbial load among seasons was observed in samples collected during spring. Escherichia coli was found in 11.57% of samples and 2.62% of isolates were found to be able to produce extended spectrum β -lactamase (ESBL). All samples were found negative for Salmonella enterica , whereas Listeria monocytogenes was present in 3.70% of samples. Higher levels of spoilage bacteria (lactic acid bacteria and Pseudomonas spp.) were detected during winter and spring. Additionally, the %CO 2 production was affected by the type of salad, while the interaction between producer and type of salad, affected total phenolic content and antioxidant activity of samples. A positive correlation of phenols and antioxidants with the presence of Staphylococcus spp., Pseudomonas spp., E. coli and Bacillus cereus was observed, suggesting that excessive handling increases microbial load and plant stress. • Ready-to-eat salads from the Cypriot market were assessed for microbiological quality and safety. • Spring is the season that ready-to-eat salads are more pronounced on microbial load. • Salmonella enterica was not found in any of the samples tested. • Polyphenols and antioxidants were positively correlated with the presence of some pathogens. • Various salad types are respiring differently through the respiration metabolic process. [ABSTRACT FROM AUTHOR]

Salmonella enterica serotypes have been reported as the agent of various outbreaks occurred after the consumption of low water activity (a w) foods. When the pathogen encounters harsh conditions, several regulatory networks are activated through dynamic differential gene expression that lead to cell survival for prolonged periods. In this work, the transcriptome of S. enterica serovar Typhimurium using RNA-Seq, after cells' inoculation in four distinct types of low a w foods (milk chocolate, powdered milk, black pepper, and dried pet food), following storage at 25 °C per 24 and 72 h was studied. The findings of this study suggest that gene regulation is influenced by the food composition mainly in the first 24 h post-inoculum, proceeded by the induction of similar genes shared among all samples. It was possible to evaluate the differences on each type of food matrix regarding the bacteria adaptation, as well as the similarities provoked by low a w. The results reveal genes that may play key roles in response to desiccation in Salmonella , as well as the pathways in which they are involved. • RNA-Seq was performed in S. enterica after inoculum in four different types of food. • Macronutrients content affects mainly the first 24 h of S. enterica adaptation. • After 72 h inoculum, dehydration responses presented higher similar patterns. • Plasmidial genes of S. enterica SL1344 were highly induced after 72 h of inoculum. [ABSTRACT FROM AUTHOR]

The replacement of the most prevalent Salmonella enterica genotypes has been documented worldwide. Here we tested the hypothesis that the current prevalent sequence type ST213 of serotype Typhimurium in Mexico has a higher resistance to stressful food preservation conditions than the displaced sequence ST19. ST19 showed higher cell viability percentages than ST213 in osmotic (685 mM NaCl) and acidic (pH 3.5) stress conditions and in combination with refrigeration (4 °C) and ambient (≈22 °C) temperatures. Both genotypes showed the same poststress recovery growth. ST213 formed biofilm and filamentous cells (FCs) under stress, whereas ST19 did not. ST213 cells also showed higher motility. The capacity of ST213 to form FCs may explain its lower viability percentages when compared with ST19, i.e., ST213 cells divided less under stress conditions, but FCs had the same recovery capacity of ST19 cells. ST213 presented a higher unsaturated/saturated fatty acids ratio (0.5–0.6) than ST19 (0.2–0.5), which indicates higher membrane fluidity. The transcript levels of the rpoS gene were similar between genotypes under the experimental conditions employed. Biofilm formation, the generation of FCs, cell motility and membrane modification seem to make ST213 more resistant than ST19 to food preservation environments. • The replacement of the predominant Salmonella enterica genotype occurs worldwide. • The sequence type ST213 is displacing ST19 as the prevalent food genotype in Mexico. • Under stress, ST213 generates filamentous cells and has higher motility than ST19. • ST213 forms biofilm and modifies membrane lipids composition under stress. • ST213 resistance to food-preservation conditions partially explains ST19 replacement. [ABSTRACT FROM AUTHOR]

To evaluate the feasibility of acidified sodium benzoate (NaB) as alternative washing solutions of fresh produce, the survival of inoculated pathogens, the background molds and yeasts counts, and quality parameters were compared during 4 and 21 °C storage of cherry tomatoes washed with 3000 ppm NaB at pH 2.0, 200 ppm free chlorine at pH 6.5, water adjusted to pH 2.0, and distilled water. The acidified NaB solution was the most effective in reducing the population of Escherichia coli O157:H7, Salmonella enterica and Listeria monocytogenes cocktails on tomatoes (>4 log CFU/g). NaB was more effective than free chlorine (P < 0.05) in reducing the two Gram-negative bacteria on tomatoes, while the reduction of Gram-positive L. monocytogenes by NaB (5.49 log CFU/g) and chlorine (4.98 log CFU/g) was similar (P > 0.05). No recovery of bacteria was found in all treatments during storage for 15 days. The acidified NaB effectively controlled yeasts and molds on cherry tomatoes to <1 log CFU/g or below the detection limit at both temperatures during 15-d storage, while free chlorine did not. Compared to unwashed controls, NaB had no effect on color, weight loss, firmness, and total soluble solids content of tomatoes during storage. The effect of NaB reducing pathogenic and spoilage microorganisms on tomatoes and maintaining quality during storage suggests its potential as an alternative wash solution in postharvest processing of fresh produce. • Effects of acidified sodium benzoate (NaB) as a washing solution were tested. • All pathogens on tomatoes were reduced by > 4 Log CFU/g with 0.3% NaB at pH 2.0. • No recovery of pathogens was observed during storage at 4 and 21 °C for 15 d. • Yeasts and molds on cherry tomatoes washed with NaB were controlled during storage. • Quality of cherry tomato was not affected by acidified NaB. [ABSTRACT FROM AUTHOR]

Abstract Controlling the presence of pathogenic bacteria, such as Salmonella sp., in dairy products production is a burning issue since contamination with Salmonella can occur at any stage of the production chain. The use of Salmonella -phages applied as control agents has gained considerable interest. Nonetheless, Salmonella -phage applications specifically intended for ensuring the safety of dairy products are scarce. This review identifies recent advances in the use of Salmonella -phages that are or could be applied along the dairy food chain, in a farm-to-fork approach. Salmonella -phages can be promising tools to reduce the shedding of Salmonella in cattle, and to reduce and control Salmonella occurrence in postharvest food (such as food additives), and in food processing facilities (such as biosanitizing agents). These control measures, combined with existing methods and other biocontrol agents, constitute new opportunities to reduce Salmonella occurrence along the dairy food production, and consequently to alleviate the risk of Salmonella contamination in dairy products. Highlights • Salmonella -phages have gained considerable interest for use as biocontrol agents. • A lack of data exists regarding the use of Salmonella -phages along the dairy food chain. • Combining phages with other biocontrol agents is an attractive option for controlling Salmonella. [ABSTRACT FROM AUTHOR]

Abstract Contamination of eggs with Salmonella enterica is a significant risk factor contributing to foodborne disease. Periods of peak egg contamination were identified by conducting longitudinal environmental and egg sampling in 7 layer flocks until they were 50 weeks of age. A total of 714 environmental samples and 8958 eggs were cultured using standard methods for the detection of salmonellae. Pooled egg contamination with Salmonella Typhimurium or Salmonella Infantis was detected at a true prevalence (TP) of 0.002 (95% CI = 0.001, 0.004) or 0.005 (95% CI = 0.004, 0.007), respectively. S. Typhimurium and S. Infantis were detected in individual egg components; in shell rinse at a TP of 0.014 (95% CI = 0.005, 0.038), in shell and membrane at a TP of 0.01 (95% CI = 0.003, 0.032), and in albumen and yolk content at a TP of 0.007 (95% CI = 0.001, 0.027). The concentration of salmonellae in all fractions was <1 CFU/mL. The TP of Salmonella enterica in eggs was highest at the onset of lay. Higher egg prevalence was associated with a lower body weight, higher egg production, higher egg weight and mass than the breed standard for age, and poorer feed conversion efficiency. Flock physiology appears to have an important influence on the detection of eggs contaminated with Salmonella enterica. Highlights • The true prevalence of Salmonella spp. in eggs was highest at the onset of lay. • S. Typhimurium and S. Infantis were detected in the internal egg content (yolk and albumen) at a true prevalence of 0.007 (95% CI: 0.001, 0.027). • Environmental contamination with Salmonella did not affect the true prevalence of the same salmonellae in eggs. • Higher egg prevalence was associated with a lower body weight, higher egg production and egg weight than the breed standard. [ABSTRACT FROM AUTHOR]

Using naturally-occurring bacterial strains as positive controls in testing protocols is typically feared due to the risk of cross-contaminating samples. We have developed a collection of strains which express Green Fluorescent Protein (GFP) at high-level, permitting rapid screening of the following species on selective or non-selective plates: Escherichia coli O157:H7, Shigella sonnei , S. flexneri , Salmonella enterica subsp. Enterica serovar Gaminera, S. Mbandaka, S. Tennesse, S. Minnesota, S. Senftenberg and S. Typhimurium. These new strains fluoresce when irradiated with UV light and maintain this phenotype in absence of antibiotic selection. Recombinants were phenotypically equivalent to the parent strain, except for S. Tennessee Sal66 that appeared Lac − on Xylose Lysine Deoxycholate (XLD) agar plates and Lac + on Mac Conkey and Hektoen Enteric agar plates. Analysis of closed whole genome sequences revealed that Sal66 had lost one lactose operon; slower rates of lactose metabolism may affect lactose fermentation on XLD agar. These fluorescent enteric control strains were challenging to develop and should provide an easy and effective means of identifying cross-contamination. [ABSTRACT FROM AUTHOR]

Bacterial transfer during postharvest handling of fresh produce provides a mechanism for spreading pathogens, but risk factors in dry environments are poorly understood. The aim of the study was to investigate factors influencing bacterial transfer between yellow onions (Allium cepa) and polyurethane (PU) or stainless steel (SS) under dry conditions. Rifampin-resistant Enterococcus faecium NRRL B-2354 or a five-strain cocktail of Salmonella was inoculated onto onion skin or PU surfaces at high or moderate levels using peptone, onion extract, or soil water as inoculum carriers. Transfer from inoculated to uninoculated surfaces was conducted using a texture analyzer to control force, time, and number of contacts. Transfer rates (ratio of recipient surface to donor surface populations) of E. faecium (4–5%) were significantly higher than those of Salmonella (0.5–0.6%) at the high (7 log CFU/cm2) but not moderate (5 log CFU/cm2) inoculum levels. Significantly higher populations of E. faecium transferred from onion to PU than from PU to onion. The transfer rates of E. faecium were impacted by inoculum carrier (61% [onion extract], 1.6% [peptone], and 0.31% [soil]) but not by inoculation level or recipient surface (PU versus SS). Bacterial transfer during dry onion handling is significantly dependent on bacterial species, inoculation levels, inoculum carrier, and transfer direction. • Transfer of E. faecium was greater from onions than from polyurethane. • Transfer rates for E. faecium were not influenced by inoculation level. • Transfer rates of Salmonella were smaller at high than at low inoculation levels. • Transfer rates increased in onion extract and decreased in soil compared to peptone. • Transfer was similar from onions to polyurethane or to stainless steel. [ABSTRACT FROM AUTHOR]

The influence of chemical diluents on the antimicrobial activity of plant essential oil (EO) vapors was evaluated. We first determined if vapors generated from 22 chemical diluents not containing EO had antimicrobial activities. Ethyl ether vapor retarded the growth of S. aureus. The minimal inhibitory concentrations (MICs) and the minimal lethal concentrations (MLCs) of cinnamon bark EO vapor, which was diluted in and generated from 21 diluents, against S. aureus and S. enterica were determined. Cinnamon bark EO vapor showed significantly (P ≤ 0.05) lower MICs against S. aureus when diluted in dimethyl sulfoxide (DMSO), ethanol, ethyl acetate, or jojoba oil, and against S. enterica when diluted in DMSO, ethanol, or jojoba oil, compared to those in other diluents. We compared antimicrobial activities of cinnamon bark EO vapor diluted in DMSO, ethanol, ethyl acetate, or jojoba oil against S. aureus and S. enterica on beef jerky as a food model. Antimicrobial activity was significantly (P ≤ 0.05) higher when vaporized from DMSO. These results indicate that antimicrobial activity of cinnamon bark EO vapor may vary significantly (P ≤ 0.05) depending on the type of diluent from which it is vaporized. These observations provide basic information when developing food and food-contact surface decontamination strategies using EO vapors. • Influence of diluent on antimicrobial effects of essential oil (EO) vapor was studied. • Antimicrobial activities of EO vapors were changed according to the type of diluents. • Antimicrobial effect of cinnamon bark EO vapor was strongest when vaporized from DMSO. • EO vapor generated from vegetable oils showed relatively lower antimicrobial activity. [ABSTRACT FROM AUTHOR]

Drug-resistant Salmonella is widely distributed in the meat production chain, endangering food safety and public health. Acidification of meat products during processing can induce acid stress, which may alter antibiotic resistance. Our study investigated the effects of acid stress on the antibiotic resistance and metabolic profile of Salmonella Typhimurium, and explored the underlying mechanisms using metabolomic and transcriptomic analysis. We found that acid-stressed 14028s was more sensitive to small molecule hydrophobic antibiotics (SMHA) while more resistant to meropenem (MERO). Metabolomic analysis revealed that enhanced sensitivity to SMHA was correlated with increased purine metabolism and tricarboxylic acid cycle. Transcriptomic analysis revealed the downregulation of chemotaxis-related genes, which are also associated with SMHA sensitivity. We also found a significant downregulation of the ompF gene, which encodes a major outer membrane protein OmpF of Salmonella. The decreased expression of OmpF porin hindered the influx of MERO, leading to enhanced resistance of the bacteria to the drug. Our findings contribute to greatly improve the understanding of the relationship between Salmonella metabolism, gene expression, and changes in drug resistance after acid stress, while providing a structural framework for exploring the relationship between bacterial stress responses and antibiotic resistance. Schematic diagram of changes in drug resistance caused by changes in intracellular purine metabolic pathway, TCA cycle and OmpF membrane porin in S. Typhimurium after acid stress. [Display omitted] • Mutlimics were used to more comprehensively reveal the drug resistance mechanism. • The acceleration of purine metabolism and TCA cycle increased the sensitivity of S. Typhimurium to SMHA. • Acid stress leads to down-regulation of ompF expression and reduced OmpF porin synthesis. • Acid stress leads to reduction of porins leading to increased resistance to meropenem. [ABSTRACT FROM AUTHOR]

Salmonella enterica is an important foodborne pathogen that constitutes a major health hazard. The emergence and aggravation of antibiotic-resistant Salmonella has drawn attention widely around the world. Conducting a risk assessment of antibiotic-resistant foodborne pathogens throughout the food chain is a pressing requirement for ensuring food safety. The growth, survival capability, and virulence of antibiotic-resistant Salmonella represent crucial biological characteristics that play an important role in microbial risk assessment. In this study, eight antibiotic-sensitive S. enterica strains were induced by Ampicillin (Amp) and Ciprofloxacin (CIP), respectively, and AMP-resistant and CIP-resistant mutants were obtained. The growth characteristics under different temperatures (25, 30, 35 °C), viability after exposure to heat (55, 57.5, 60 °C) and acid (HCl, pH = 3.0), the virulence potential (adhesion and invasion to Caco-2 cells, biofilm formation and motility) and the lethality in a model species (Galleria mellonella) were evaluated and compared for S. enterica strains before and after antibiotic exposure. The induction by AMP and CIP are likely to promote cross-antibiotic resistance to their antibiotic classes, β-lactams and quinolones, as well as some compound antibiotics. It was observed that generally the antibiotic-induction-resistant strains showed decreased growth ability and lower heat resistance, although the differences were not significant at all the conditions tested. The AMP-resistant strains were significantly less acid resistance than the sensitive and the CIP-resistant ones, while exhibiting increased biofilm formation ability. In general, the antibiotic-induced resistance did not significantly affect the motility, adherence, or invasion ability of Caco-2 cells. However, CIP-resistant strains displayed lower lethality in G. mellonella infection, whereas AMP-resistant strains did not, and even two strains improved lethality. The study of the biological characteristics of antibiotic-resistant S. enterica is essential in better understanding the microbial risks to both the food chain and human health, thereby facilitating a more accurate risk assessment. • AMP/CIP induce cross-antibiotic resistance and decrease growth/heat resistance of Salmonella. • G. mellonella lethality correlates with Caco-2 invasion ability. • Two AMP-induced strains exhibit increased G. mellonella lethality. [ABSTRACT FROM AUTHOR]

Lawn-harvest method uses a solid medium (e.g., tryptic soy agar, TSA) to produce bacterial lawns and is widely accepted for the culture of microorganisms in microbial studies of low-moisture foods (LMFs, foods with water activity less than 0.85). It produces desiccation-tolerant cells with higher D-values in LMFs; however, little is known about the molecular mechanisms underlying bacterial resistance. Salmonella enterica Enteritidis PT 30 (S. Enteritidis), the most pertinent pathogen in LMFs, was cultured in TSA and tryptic soy broth (TSB). Cells were harvested and inoculated on filter papers to assess their performance under a relative humidity of 32 ± 2%. Transcriptome analysis of cultured cells during long-term desiccation (24, 72, and 168 h) was conducted in TruSeq PE Cluster Kit (Illumina) by paired-end methods. Lawn-cultured S. Enteritidis cells have stronger survivability (only decreased by 0.78 ± 0.12 log after 130 d of storage) and heat tolerance (higher D/β value) than those from the broth method. More desiccation genes of lawn-cultured cells were significantly upregulated from growth to long-term desiccation. Differentially expressed genes were the most enriched in the ribosome and sulfur metabolism pathways in the lawn- and broth-cultured groups. This study tracked the transcriptomic differences between two cultured groups in response to long-term desiccation stress and revealed some molecular mechanisms underlying their different suitability in microbial studies of LMFs. • Bacteria gain desiccation tolerance during their growth on lawns. • Ribosome metabolism of lawn-cultured cells was most enriched during desiccation. • Broth-cultured cells had a non-linear survivor curve at 85 °C relative humidity of 32%. [ABSTRACT FROM AUTHOR]

Two specialty mushrooms have recently become novel vectors for foodborne outbreaks in the U.S.: fresh enoki and dried wood ear mushrooms were linked to a listeriosis and salmonellosis outbreak, respectively. The aim of this study was to evaluate the survival kinetics of Listeria monocytogenes and Salmonella enterica on dehydrated enoki and wood ear mushrooms during long-term storage. Following heat dehydration, mushrooms were inoculated with either L. monocytogenes or S. enterica , allowed to dry for 1 h, and then stored for up to 180 d at 25 °C and 33% relative humidity. Both pathogens were enumerated from the mushrooms at intervals during the storage period. Survival kinetics of both pathogens were modeled using both the Weibull and log-linear with tail models. After inoculation and 1 h drying, both pathogen populations decreased 2.26–2.49 log CFU/g on wood ear mushrooms; no decrease was observed on enoki. Both pathogens survived during storage on both mushroom types. On wood ear mushrooms, a 2-log decrease of both pathogens occurred during storage. On enoki mushrooms, 4-log decreases of both pathogens were modeled to occur after 127.50–156.60 d. The results of this study suggest that L. monocytogenes and S. enterica can persist on dehydrated specialty mushrooms during long-term storage. • Both pathogens survived on dehydrated enoki and wood ear mushrooms during storage. • Both pathogens decreased on dehydrated wood ear mushrooms after inoculation. • The Weibull and log-linear with tail models adequately described survival kinetics. • Reductions of 4 log CFU/g were observed on enoki mushrooms during storage. • Storage alone did not eliminate either pathogen on dehydrated mushrooms. [ABSTRACT FROM AUTHOR]

Salmonella enterica serovar Heidelberg ( S. Heidelberg) is one of the top serovars causing human salmonellosis. This serovar ranks second and third among serovars that cause human infections in Québec and Canada, respectively, and has been associated with severe infections. Traditional typing methods such as PFGE do not display adequate discrimination required to resolve outbreak investigations due to the low level of genetic diversity of isolates belonging to this serovar. This study evaluates the ability of four whole genome sequence (WGS)-based typing methods to differentiate among 145  S. Heidelberg strains involved in four distinct outbreak events and sporadic cases of salmonellosis that occurred in Québec between 2007 and 2016. Isolates from all outbreaks were indistinguishable by PFGE. The core genome single nucleotide variant (SNV), core genome multilocus sequence typing (MLST) and whole genome MLST approaches were highly discriminatory and separated outbreak strains into four distinct phylogenetic clusters that were concordant with the epidemiological data. The clustered regularly interspaced short palindromic repeats (CRISPR) typing method was less discriminatory. However, CRISPR typing may be used as a secondary method to differentiate isolates of S. Heidelberg that are genetically similar but epidemiologically unrelated to outbreak events. WGS-based typing methods provide a highly discriminatory alternative to PFGE for the laboratory investigation of foodborne outbreaks. [ABSTRACT FROM AUTHOR]

We examined the survival of Listeria monocytogenes , Escherichia coli O157:H7, and Salmonella enterica Thompson inoculated on commercially packed table grapes under simulated refrigerated transit conditions (1.1 ± 0.5 °C; 90% RH). Grapes were placed in perforated polyethylene cluster bags, within a commercial expanded polystyrene box equipped with either a SO 2 -generating pad; a perforated polyethylene box liner; a SO 2 -generating pad and a box liner; or none of them. L. monocytogenes was most sensitive to SO 2 -generating pad. SO 2 -generating pad or SO 2 -generating pad with box liner inactivated this pathogen completely on day 12 following the inoculation. S. enterica Thompson displayed a similar cold sensitivity as L. monocytogenes, but was more resistant to SO 2 -generating pad than L. monocytogenes . While SO 2 -generating pad eliminated S. enterica Thompson on day 20, a combination of box liner with SO 2 -generating pad inactivated this pathogen completely on day 13. E. coli O157:H7 had the highest tolerance to transit temperature and to SO 2 -generating pad; SO 2 -generating pad inactivated this pathogen completely on Day 20. Our data suggest that use of SO 2 -generating pad combined with box liner is effective in reducing foodborne pathogens L. monocytogenes and S. enterica Thompson, while the use of SO 2 -generating pad alone was more effective on E. coli O157:H7. [ABSTRACT FROM AUTHOR]

This study was conducted to determine the effects of intrinsic juice characteristics namely insoluble solids (IS, 0–3 %w/v), and soluble solids (SS, 0–70 °Brix), and extrinsic process parameter treated volume (250–1000 mL) on the UV-C inactivation rates of heat-stressed Salmonella enterica in simulated fruit juices (SFJs). A Rotatable Central Composite Design of Experiment (CCRD) was used to determine combinations of the test variables, while Response Surface Methodology (RSM) was used to characterize and quantify the influences of the test variables on microbial inactivation. The heat-stressed cells exhibited log-linear UV-C inactivation behavior (R 2 0.952 to 0.999) in all CCRD combinations with D UV-C values ranging from 10.0 to 80.2 mJ/cm 2 . The D UV-C values obtained from the CCRD significantly fitted into a quadratic model (P < 0.0001). RSM results showed that individual linear (IS, SS, volume), individual quadratic (IS 2 and volume 2 ), and factor interactions (IS × volume and SS × volume) were found to significantly influence UV-C inactivation. Validation of the model in SFJs with combinations not included in the CCRD showed that the predictions were within acceptable error margins. [ABSTRACT FROM AUTHOR]

Non-pathogenic bacterial biofilms were developed on the surface of stainless steel possessing desiccation tolerance and antimicrobial activity against Salmonella enterica . Three bacteria exhibiting strong antimicrobial activities against S. enterica were isolated from various soils, foods, and food-contact surfaces. Isolates were identified as Pseudomonas extremorientalis (strain Lettuce-28), Paenibacillus peoriae (strain Lettuce-7), and Streptomyces cirratus (strain Geumsan-207). These bacteria grew rapidly and formed biofilms within 24 h on the surface of stainless steel coupons (SSCs) immersed in laboratory media (tryptic soy broth or Bennet's broth) at 25 °C. Cells in biofilms had enhanced tolerance to desiccation (exposure to 43% atmospheric relative humidity [RH]) and retained antimicrobial activity against S. enterica . Populations of S. enterica deposited on SSCs containing biofilm formed by Ps. extremorientalis strain Lettuce-28, for example, decreased by > 2.5 log CFU/coupon within 24 h at 25 °C and 43% RH, while the number of cells inoculated on SSCs lacking biofilm decreased by 1.5 log CFU/coupon. Antimicrobial activities of the three antagonistic bacteria against S. enterica persisted in desiccated biofilms. This study provides insights to developing strategies to inactivate Salmonella and perhaps other foodborne pathogens on abiotic surfaces using non-pathogenic antagonistic bacteria. [ABSTRACT FROM AUTHOR]

Despite the development of numerous predictive microbial inactivation models, a model focusing on the variability in time to inactivation for a bacterial population has not been developed. Additionally, an appropriate estimation of the risk of there being any remaining bacterial survivors in foods after the application of an inactivation treatment has not yet been established. Here, Gamma distribution, as a representative probability distribution, was used to estimate the variability in time to inactivation for a bacterial population. Salmonella enterica serotype Typhimurium was evaluated for survival in a low relative humidity environment. We prepared bacterial cells with an initial concentration that was adjusted to 2 × 10 n colony-forming units/2 μl (n = 1, 2, 3, 4, 5) by performing a serial 10-fold dilution, and then we placed 2 μl of the inocula into each well of 96-well microplates. The microplates were stored in a desiccated environment at 10–20% relative humidity at 5, 15, or 25 °C. The survival or death of bacterial cells for each well in the 96-well microplate was confirmed by adding tryptic soy broth as an enrichment culture. The changes in the death probability of the 96 replicated bacterial populations were described as a cumulative Gamma distribution. The variability in time to inactivation was described by transforming the cumulative Gamma distribution into a Gamma distribution. We further examined the bacterial inactivation on almond kernels and radish sprout seeds. Additionally, we described certainty levels of bacterial inactivation that ensure the death probability of a bacterial population at six decimal reduction levels, ranging from 90 to 99.9999%. Consequently, the probability model developed in the present study enables us to estimate the death probability of bacterial populations in a desiccated environment over time. This probability model may be useful for risk assessment to estimate the amount of remaining bacteria in a given sample. [ABSTRACT FROM AUTHOR]

Non-Typhoidal Salmonella (NTS) continues to be a leading cause of foodborne illness worldwide. Food manufacturers implement hurdle technology by combining more than one approach to control food safety and quality, including preservatives such as organic acids, refrigeration, and heating. We assessed the variation in survival in stresses of genotypically diverse isolates of Salmonella enterica to identify genotypes with potential elevated risk to sub-optimal processing or cooking. Sub-lethal heat treatment, survival in desiccated conditions and growth in the presence of NaCl or organic acids were investigated. S. Gallinarum strain 287/91 was most sensitive to all stress conditions. While none of the strains replicated in a food matrix at 4 °C, S. Infantis strain S1326/28 retained the greatest viability, and six strains exhibited a significantly reduced viability. A S. Kedougou strain exhibited the greatest resistance to incubation at 60 °C in a food matrix that was significantly greater than S. Typhimurium U288, S Heidelberg, S. Kentucky, S. Schwarzengrund and S. Gallinarum strains. Two isolates of monophasic S. Typhimurium, S04698–09 and B54Col9 exhibited the greatest tolerance to desiccation that was significantly more than for the S. Kentucky and S. Typhimurium U288 strains. In general, the presence of 12 mM acetic acid or 14 mM citric acid resulted in a similar pattern of decreased growth in broth, but this was not observed for S. Enteritidis, and S. Typhimurium strains ST4/74 and U288 S01960-05. Acetic acid had a moderately greater effect on growth despite the lower concentration tested. A similar pattern of decreased growth was observed in the presence of 6% NaCl, with the notable exception that S. Typhimurium strain U288 S01960-05 exhibited enhanced growth in elevated NaCl concentrations. • Food chain-related stress-tolerance of Salmonella enterica is genotype-specific. • Variation was observed for strains from distinct serovar phylogroups and strains from subclades of the same serovar phylogroup. • S. Gallinarum strain 287/91 was most sensitive to all food-chain related stress conditions included in the study. • Strains of monophasic S. Typhimurium ST34 and a S. Kedougou strain exhibited the greatest tolerance to stress. • Salmonella enterica strains were unable to replicate and survival varied during storage in a food matrix at 4 °C. [ABSTRACT FROM AUTHOR]

Chlorine treatment is the most common disinfection method in food-related environments. In addition to being simple and inexpensive, this method is very effective if used properly. However, insufficient chlorine concentrations only cause a sublethal oxidative stress in the bacterial population and may alter the growth behavior of stressed cells. In the present study, the effect of sublethal chlorine stress on the biofilm formation characteristics of Salmonella Enteritidis was evaluated. Our results demonstrated that, sublethal chlorine stress (350 ppm total chlorine) activates the biofilm (csgD , agfA , adrA and bapA) and quorum-sensing (sdiA and luxS) related genes in planktonic cells of S. Enteritidis. The higher expression of these genes illustrated that the chlorine stress induced the initiation of the biofilm formation process in S. Enteritidis. Results of the initial attachment assay confirmed this finding. In addition, the number of chlorine-stressed biofilm cells was significantly higher than non-stressed biofilm cells after 48 h incubation at 37 °C. In S. Enteritidis ATCC 13076 and S. Enteritidis KL19, the number of chlorine-stressed biofilm cells were 6.93 ± 0.48 and 7.49 ± 0.57 log CFU/cm2, while the number of non-stressed biofilm cells were 5.12 ± 0.39 and 5.63 ± 0.51 log CFU/cm2, respectively. These findings were confirmed by measurements of the major components of biofilm, i.e., eDNA, protein and carbohydrate. The amount of these components in 48-h biofilms was higher when the cells were initially subjected to sublethal chlorine stress. However, the up-regulation of the biofilm and quorum sensing genes was not observed in 48-h biofilm cells, indicating that the effect of chlorine stress had vanished in the subsequent generations of Salmonella. In total, these results revealed that sublethal chlorine concentrations can promote the biofilm-forming ability of S Enteritidis. • Biolfim formation of S. Enteritidis was promoted by chlorine-stress. • Biofilm and quorum sensing genes were up-regulated by sublethal chlorine stress. • Chlorine stress induced the initiation of biofilm formation in S. Enteritidis. • Protein, carbohydrate and eDNA contents of the biofilms were increased. [ABSTRACT FROM AUTHOR]

This study investigated the effectiveness of a polyvinyl alcohol (PVA) film containing the natural antimicrobial oregano essential oil (OEO) as an active packaging application for decreasing the microbial growth. The film exerted an antimicrobial effect via the atmosphere surrounding the food rather than direct contact, thereby preserving the quality of cherry tomatoes. A packaging film containing microencapsulated OEO was developed. The loading content increased gradually (104.29–234.29 μg OEO/mg film) with the amount of OEO incorporated (1%, 2%, and 3%), where the PVA films containing 2% OEO had the highest loading efficiency (91.64%), followed by 1% OEO (90.96%) and 3% OEO (88.38%). The antimicrobial activities of the films were evaluated by applying it to fresh cherry tomatoes at 4 °C and 22 °C for 7 days. The large 2% OEO film as well as both the small and large 3% OEO films had strong antimicrobial effects against Salmonella enterica , molds and yeasts, and mesophilic aerobic bacteria. The changes in the hardness, weight, and color of the cherry tomatoes during storage did not differ significantly. The films could be utilized as a packaging material for fresh produce with antimicrobial effects because of the controlled atmosphere surrounding the food rather than by direct contact. [ABSTRACT FROM AUTHOR]

Salmonella enterica is a well-known pathogen commonly acquired from the consumption of contaminated food. It has been estimated to affect millions of humans and cause hundreds of thousands of deaths per year globally. Pork, one of the most commonly consumed meats worldwide, has been identified as one of the main sources of human salmonellosis. In this study, we aimed to detect and characterize S. enterica from slaughtered swine and generate antimicrobial resistance profiles of select isolates. Tonsils and jejunum with mesenteric lymph nodes (MLN) were collected from a total of 240 swine from eight abattoirs (five accredited and three locally registered abattoirs) across Metro Manila. S. enterica were isolated using conventional culture methods and confirmed by PCR amplification of the invA gene. Isolates were further characterized based on somatic antigen by multiplex PCR. We report that there is no significant difference (P = 0.42) between the incidences of S. enterica in swine slaughtered in accredited (44.0%) and in locally registered abattoirs (46.7%). Most samples were contaminated with S. enterica under serogroup O:3,10. Antimicrobial susceptibility testing of 183 isolates using the VITEK ® 2 system revealed high resistance to ampicillin (67.8%) and trimethoprim/sulfamethoxazole (80.3%). Multidrug-resistance was found in 124 (67.8%) isolates. [ABSTRACT FROM AUTHOR]

Enterobacteriaceae comprise food spoilage organisms as well as food-borne pathogens including Escherichia coli . Heat resistance in E. coli was attributed to a genomic island called the locus of heat resistance (LHR). This genomic island is also present in several other genera of Enterobacteriaceae , but its function in the enteric pathogens Salmonella enterica and Enterobacter cloacae is unknown. This study aimed to determine the frequency of the LHR in food isolates of E. coli, and its influence on heat resistance in S. enterica and Enterobacter spp. Cell counts of LHR-positive strains of E. coli , S. enterica and E. cloacae were reduced by less than 1, 1, and 4 log (cfu/mL), respectively, after exposure to 60 °C for 5 min, while cell counts of LHR-negative strains of the same species were reduced by more than 7 log (cfu/mL). Introducing an exogenous copy of the LHR into heat-sensitive enteropathogenic E. coli and S. enterica increased heat resistance to a level that was comparable to LHR-positive wild type strains. Cell counts of LHR-positive S. enterica were reduced by less than 1 log(cfu/mL) after heating to 60 °C for 5 min. Survival of LHR-positive strains was improved by increasing the NaCl concentration from 0 to 4%. Cell counts of LHR-positive strains of E. coli and S. enterica were reduced by less than 2 log (cfu/g) in ground beef patties cooked to an internal core temperature of 71 °C. This study indicates that LHR-positive Enterobacteriaceae pose a risk to food safety. [ABSTRACT FROM AUTHOR]

The objective of this study was the assessment of the stationary-phase, low-pH-inducible acid tolerance response (ATR) of different Salmonella enterica strains. For this purpose, 30 strains of the pathogen were grown in tryptone soy broth in the absence (non-adapted cultures) and presence (1% w/v; acid-adapted cultures) of glucose, and then subjected to 4-h acid challenge trials at pH 3.0. Surviving populations of each strain were determined at 1-h intervals, and the Weibull model was fitted to the derived microbiological data. Extensive variability in the acid stress responses of the tested S. enterica strains was observed, with the total population reductions (log CFU/ml) attained in 4 h of acid challenge ranging from 0.9 to 5.5 and from 0.6 to 7.0 for the non-adapted and acid-adapted cultures, respectively. As demonstrated by the model scale parameter δ and shape parameter p , the effect of acid adaptation on the inactivation curves was strain-specific. Although acid adaptation resulted in enhanced acid survival for the majority of the tested strains, there were strains exhibiting similar or decreased acid resistance compared to their non-adapted counterparts. Moreover, acid adaptation appeared to decrease the strain variability of δ whereas increasing the strain variability of p : the coefficient of variation of δ among the tested strains was 97.2 and 54.9% for the non-adapted and acid-adapted cultures, respectively, while the corresponding values for p were 12.7 and 48.1%. The data of the present study, which is the first one to systematically evaluate the adaptive ATR of multiple S. enterica strains, clearly demonstrate that this phenotype (attempted to be induced by growing the pathogen in the presence of glucose) is strain-dependent. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate the lethal effects of simultaneous treatments with gaseous chlorine dioxide (ClO 2 ) and mild wet heat (55 °C at 100% relative humidity [RH]) on Salmonella enterica in chicken feces on the surface of eggshells. Gaseous ClO 2 production decreased significantly ( P ≤ 0.05) as the RH (23, 43, 68, 85, and 100%) at 25 °C was increased. The lethality of gaseous ClO 2 against S. enterica in feces on eggshells increased significantly ( P ≤ 0.05) as RH increased. For example, when treated with gaseous ClO 2 at 85 and 100% RH at 25 °C, S. enterica (5.9 log CFU/egg) was inactivated within 4 h. In contrast, at 23, 43, and 68% RH, the pathogen remained at 5.1, 5.0, and 2.8 log CFU/egg, respectively, after 6 h. Finally, when eggshells surface-contaminated with S. enterica (5.8 log CFU/egg) were treated with gaseous ClO 2 (peak concentration of ClO 2 : 185.6 ppm) at 100% RH and 55 °C, inactivation occurred within 1 h. These results indicate that treatment of surface-contaminated shell eggs with gaseous ClO 2 at elevated RH and temperature is effective in inactivating S. enterica . These observations will be useful when developing an effective sanitation program to enhance the microbiological safety of shell eggs. [ABSTRACT FROM AUTHOR]

Many niche-dependent barriers along the poultry production continuum favour the survival of certain Salmonella serovars over others. Historically, the presence of particular serovars has been determined by niche-specific genes which encode resistance to selective pressures such as host defenses and industrial antimicrobial practices. Over the past decade, Canada has witnessed unexplained shifts in the Salmonella landscape in the poultry sector. Several formerly minor Salmonella serovars, including S. Kentucky and S. Reading, have recently increased in prevalence in live chickens and turkeys, respectively, in British Columbia (BC). The purpose of this research was to investigate the genomic features of the top poultry-associated Salmonella spp. in BC, to probe for serovar-specific characteristics that could address the recently shifting balance of serovars along the poultry continuum. By examining the quantity and diversity of antimicrobial resistance (AMR) genes, virulence factors (VFs), Salmonella Pathogenicity Islands (SPIs), and plasmids across 50 poultry-associated S. enterica isolates using whole genome sequencing and antimicrobial resistance profiling, we have identified serovar-specific differences that likely influence niche survival. Specifically, isolates in our collection from predominantly human pathogenic serovars (S. I 4, [5], 12:i: , S. Typhimurium, and S. Enteritidis) were found to share the IncFIB(S) and IncFII(S) plasmids which carry important VFs known to aid in human host infection. Additionally, these strains held the lowest number of AMR genes, and the highest number of unique SPIs which also facilitate virulence. However, other serovars containing a greater diversity and abundance of resistance genes have been increasing across the poultry sector. S. Kentucky was found to carry unique AMR genes, VFs, SPIs, and plasmids that could bolster persistence in farm and processing environments. Overall, S. Kentucky also had comparatively high levels of intra-serovar genetic variability when compared to other prominent serovars from our collection. In addition, one of our two S. Reading isolates had high carriage of both AMR genes and VFs relative to other isolates in our collection. As the poultry-associated Salmonella landscape continues to evolve in Canada, future studies should monitor the genetic composition of prominent serovars across poultry production to maintain up-to-date risk assessments of these foodborne pathogens to consumers. • Serovars expanding in poultry production carry more resistance genes. • Kentucky demonstrated the highest genetic diversity among observed serovars. • Enteritidis, Typhimurium, Rissen, and Brandenburg were susceptible to tested antimicrobials. [ABSTRACT FROM AUTHOR]

The photodynamic inactivation (PDI) uses molecules (photosensitizers) that absorb visible light (385–450 nm) energy, transfer it to adjacent molecular oxygen and thereby generating the biocidal singlet oxygen and other reactive oxygen species in situ. Efficacy of PDI was tested against Listeria monocytogenes and Salmonella enterica in three ways. Firstly, by adding the photosensitizer to bacterial suspensions. Secondly, bacteria were placed on inanimate surfaces and then sprayed with a photosensitizer suspension. Thirdly, bacteria were placed on coated inanimate surfaces, where the photosensitizer was permanently fixed in this coating (antimicrobial coating, AMC). Experiments were performed without and with soiling (albumin, sheep erythrocytes). In suspension, PDI reduced the number of viable Listeria monocytogenes and Salmonella enterica by more than 6 Log CFU/mL within seconds of light exposure. Photosensitizer spray suspension reduced the bacterial burden on surfaces with up to about 6 Log CFU/mL (5 s light exposure). PDI, even in the presence of high soiling, achieved a reduction of up to 5.1 ± 1.2 Log CFU/mL. The AMC showed a bacterial reduction that decreased from 5.1 to 0.7 Log CFU/mL with increasing soiling. Depending on the soiling and the respective bacteria, the spray suspension or AMC achieved a bacterial reduction on the running conveyor belt demonstrator ranging from 2.9 to 5.3 or 0.5 to 4.5 Log CFU/mL, respectively. PDI used visible light, phenalene-1-one and curcumin photosensitizers, and oxygen from ambient air to reduce the bioburden on typical surfaces in food processing. The AMC acts slower than the spray suspension but enables a permanent, self-sanitizing effect. • Photodynamic Inactivation (PDI) generates reactive oxygen species that destroys bacteria cells via oxidation. • bactericidal effect of PDI was evaluated in suspension, as spray suspension on contaminated surfaces and as antimicrobial coating. • The viability of L. monocytogenes and S. enterica in suspension was reduced by up to 6 Log CFU/mL. • Spray suspension and antimicrobial coating destroyed these bacteria also on inanimate surfaces by up to 5.8 Log CFU/mL. [ABSTRACT FROM AUTHOR]

Salmonella enterica is a ubiquitous and multi-host pathogen that causes significant morbidity and mortality worldwide. Outbreaks of foodborne salmonellosis continue to occur, highlighting the need for additional interventions. The present study investigated the potential for the commercial protective culture Hafnia alvei B16 to provide enhanced protection against multi-drug resistant strains of S. enterica serovars Typhimurium and Newport by attenuating their virulence when cocultured in milk (as a model food) and broth, and by protecting intestinal epithelial cells from pathogen infection in vitro. Exposure to HA in milk inhibited the subsequent adhesion of S. Typhimurium by 95.23%, whereas the invasion capacity of both serovars was reduced when cocultured with HA in broth and milk. The inhibition of invasion by S. Typhimurium and S. Newport was greater when cocultured in milk (86.95% and 86.58%, respectively) compared to broth (51.64% and 79.88%, respectively). Exposure to HA in both media decreased the expression of virulence genes in S. Typhimurium and S. Newport. Pre-treatment of Caco-2 cells with HA reduced invasion of S. Newport by 89.68% compared to control. These data demonstrate the potential for HA to enhance food safety by attenuating Salmonella virulence and protecting against pathogen invasion of intestinal epithelial cells. • Coculture with B16 in milk reduced S. Typhimurium adhesion to human Caco-2 cells. • Coculture with B16 reduced subsequent Salmonella invasion of human Caco-2 cells. • Pre-treatment of Caco-2 cells with B16 reduced invasion of S. Newport. • Coculture with B16 decreased the expression of virulence genes in Salmonella. [ABSTRACT FROM AUTHOR]

We investigated a bacterial sample preparation procedure for single-cell studies. In the present study, we examined whether single bacterial cells obtained via 10-fold dilution followed a theoretical Poisson distribution. Four serotypes of Salmonella enterica , three serotypes of enterohaemorrhagic Escherichia coli and one serotype of Listeria monocytogenes were used as sample bacteria. An inoculum of each serotype was prepared via a 10-fold dilution series to obtain bacterial cell counts with mean values of one or two. To determine whether the experimentally obtained bacterial cell counts follow a theoretical Poisson distribution, a likelihood ratio test between the experimentally obtained cell counts and Poisson distribution which parameter estimated by maximum likelihood estimation (MLE) was conducted. The bacterial cell counts of each serotype sufficiently followed a Poisson distribution. Furthermore, to examine the validity of the parameters of Poisson distribution from experimentally obtained bacterial cell counts, we compared these with the parameters of a Poisson distribution that were estimated using random number generation via computer simulation. The Poisson distribution parameters experimentally obtained from bacterial cell counts were within the range of the parameters estimated using a computer simulation. These results demonstrate that the bacterial cell counts of each serotype obtained via 10-fold dilution followed a Poisson distribution. The fact that the frequency of bacterial cell counts follows a Poisson distribution at low number would be applied to some single-cell studies with a few bacterial cells. In particular, the procedure presented in this study enables us to develop an inactivation model at the single-cell level that can estimate the variability of survival bacterial numbers during the bacterial death process. [ABSTRACT FROM AUTHOR]

The objective of the present study was to compare the growth of food-pathogens Listeria monocytogenes , Salmonella enterica, food spoilage Bacillus subtilis , an industrial milk product isolate, and spore-forming Paenibacillus in commercially available ultrahigh temperature processed (UHT) bovine milk and non-dairy, plant-based beverages (coconut, almond, cashew) stored at chilled and ambient temperatures (4 °C, 8 °C or 20 °C). Beverage samples were inoculated with a strain cocktail or individual strains of either Listeria or Salmonella, or Paenibacillus or Bacillus , respectively (approximately 1 × 103 CFU/mL). The findings indicate that the bacterial strains used in the study were capable of proliferating in plant-based beverages at higher rates than in bovine milk at 8 °C and 20 °C for Listeria and 20 °C for Salmonella and Paenibacillus , respectively. Bacillus subtilis grew equally fast in bovine milk and plant-based almond drink at 20 °C. No statistically significant difference (p > 0.05) in growth rates between different types of tested beverages was observed at 4 °C and at 8 °C for Listeria and Salmonella cocktails, respectively. These data suggest that plant-based beverages may present a significant risk for listeriosis and salmonellosis and post-opening recommendations should be carefully considered. • Plant-based milk drinks support rapid growth of Listeria, Salmonella, Bacillus and Paenibacillus. • Listeria grew significantly faster in plant-based beverages than in bovine milk at 8 °C and 20 °C. • Salmonella growth rate was significantly higher in plant-based beverages than bovine milk at 20 °C. • Growth of sporeformers was equivalent or faster in plant-based beverages than bovine milk at 20 °C. [ABSTRACT FROM AUTHOR]

Salmonella enterica serovar Typhimurium monophasic variants (Salmonella 4,[5],12:i:-) has increased dramatically, causing human salmonellosis and colonization in pigs. With a difference to S. Typhimurium, the monophasic variants of S. Typhimurium lose the gene cassettes encoding the second phase flagellin. To establish a rapid method to detect and differentiate the two serotypes, we analyzed the published 679 genomes of S. Typhimurium and its monophasic variants and found that no Salmonella 4,[5],12:i:- strains carry both fljB and hin genes. Therefore, we established a novel multiplex PCR method using the fljB-hin region and mdh gene as target sequences to detect and differentiate both serotypes. This method can be used to specifically detect both serotypes with a detection limit for DNA concentration at 10 pg/μL. In addition, the PCR assay successfully differentiated 36 S. Typhimurium isolates from 62 isolates of monophasic variants preserved in our laboratory from 2009 to 2017, which corresponds to the whole-genome-based serotyping results. Application of the multiplex PCR method to 60 fecal samples from a pig farm identified 11.7% (7/60) of S. Typhimurium monophasic variants, which is consistent with the whole-genome-based serotyping results. The multiplex PCR assay is a rapid and precise method for the detection of S. Typhimurium monophasic variants from samples across food production chains. • Genetic difference between S. Typhimurium and its monophasic variant was identified by comparative genomics. • No Salmonella 1,4,[5],12:i:- strains carry both fljB and hin genes. • A new multiplex PCR assay was established to differentiate S. Typhimurium from Salmonella 1,4,[5],12:i:-. • 11.7% of Salmonella 1,4,[5],12:i:- was detected in a pig farm using the PCR assay. [ABSTRACT FROM AUTHOR]

Salmonella spp. is one of the leading causes of foodborne outbreaks worldwide. Salmonella spp. has been associated with a variety of food sources, particularly egg products. They can enter a viable but nonculturable (VBNC) state in response to harsh stress. VBNC cells still retain membrane integrity and metabolic activity, which may pose health risks. However, the formation mechanism and resuscitation ability of VBNC cells are not well understood. In this work, Salmonella spp. cocktails, including Salmonella enterica serovar Newport and Salmonella enterica serovar Enteritidis, in liquid egg products was induced into a VBNC state by mild heat treatment, a commonly used method to inhibit the growth of pathogenic in liquid egg industry. Mild heat induced VBNC cells were found to resuscitate in liquid egg yolk (LEY) and liquid whole egg (LWE), but they failed to recover in liquid egg white (LEW). In addition, a certain number of cells remained as VBNC state after in vitro digestion. The membrane vesicle (MV) protein encoding gene pagC , two-component system encoding genes phoP/Q and sigma factor encoding gene rpoS were highly expressed in VBNC cells compared with the culturable counterparts. The results of this study can contribute to a better understanding of the health risks associated with Salmonella spp. in VBNC state and provide a theoretical basis for formation mechanism of VBNC state. • Salmonella spp. in liquid egg products entered into the VBNC state under mild heat. • VBNC cells were resuscitated in liquid egg yolk and liquid whole egg stored at 25 °C. • A certain number of cells remained as VBNC state after in vitro digestion. • PagC gene its regulatory genes were highly expressed in VBNC state cells. [ABSTRACT FROM AUTHOR]

Plants influence epiphytic bacterial associations but Salmonella enterica colonizes crop plants commensally, raising the possibility of human foodborne illness, but the factors that mediate human pathogen-plant associations remain understudied. We evaluated whether any changes in leaf tissue and surface metabolomes with kale (Brassica oleracea Acephala group) development and in response to drought modulated Salmonella leaf association. Untargeted phytochemical profiling (including primary and secondary metabolites) of kale leaf tissue extracts and leaf surface washes revealed distinct metabolite profiles that shifted with plant development. Metabolomes of juvenile plants also diverged in response to drought stress, an effect not noted in mature kale. Restricted watering in juvenile plants led to up-accumulation of 45 compounds in leaf tissue and 21 in leaf wash and the appearance of several unique peaks, with concomitant increases in phytochemical measurements. The antioxidant capacity and total flavonoid content were higher in mature than juvenile, regularly watered plant leaf extracts. Drought also elicited flavonoids and glucosinolates in juvenile plants. In mature plants, drought did not induce further prominent changes. Regularly watered juvenile kale provided a favorable substrate for inoculated Salmonella but the ability to support Salmonella declined with age and with drought stress. Salmonella growth was impaired in mature or water-stressed plant washes compared to controls and positive correlations were detected between Salmonella counts on leaves and in leaf washes. Moreover, Salmonella counts were inversely correlated with total flavonoids and phenolics in kale tissues from juvenile plants and regularly watered plants. Future studies should assess how changes in primary and secondary metabolites on the kale plant surface can modulate the Salmonella association. Regulated water restriction could be a strategy in controlled agriculture, with the dual purpose of enhancing health beneficial quality and food safety, especially when harvested at the baby kale stage. [Display omitted] • Flavonoids and phenolics were higher in mature plants and water-stressed baby kale. • Age negatively affected Salmonella association with regularly watered kale. • Drought restricted Salmonella survival on juvenile but not mature kale plants. • Counts of Salmonella associating epiphytically correlated with counts in leaf wash. • Salmonella counts were inversely correlated with flavonoids and phenolics in kale. [ABSTRACT FROM AUTHOR]

Cross-protection represents a considerable challenge in the food industry where hurdled interventions are often employed to reduce Salmonella contamination. The heat resistance of Salmonella strains from five serotypes (i.e., Typhimurium, Enteritidis, Tennessee, Thompson and Hartford) at 70 °C was determined by measurement of viable cell populations before and after adaptation to two common stresses employed in low-water activity food processing, desiccation and sub-lethal heat treatment. Survival of Salmonella at 70 °C significantly increased (p < 0.05) following the six-day incubation in peanut oil (a w 0.52 ± 0.00) and/or the exposure to a sub-lethal heat treatment at 45 °C for 3 min. Quantitative PCR revealed upregulation of two desiccation stress-related genes, fadA and otsB , following the peanut oil incubation, whereas heat treatment induced upregulation of a heat-resistance gene, dnaK . Invasion gene invA and alternative sigma factor rpoE were downregulated following either of the treatments. Interestingly, different Salmonella strains yielded different transcriptional profiles. The strain-specific resistance phenotypes and transcriptional profiles provided further insights into the mechanisms employed to tolerate desiccation and heat stresses in the food industry. [ABSTRACT FROM AUTHOR]

In this study, we examined the antimicrobial effects of citrus extract (Citrox ® ) and chitosan on lactic acid bacteria (LAB) and the pathogens Escherichia coli O157:H7 and Salmonella enterica on turkey meat during storage under vacuum packaging (VP) at 4 and 10 °C. We also examined the effects of Citrox and chitosan on pathogen contamination in tryptic soy broth (TSB). Chitosan alone or in combination with Citrox inhibited the growth of endogenous LAB in turkey meat, whereas citrus extract did not cause a major reduction in bacterial density. Citrus extract combined with chitosan yielded the lowest mesophilic total viable counts (TVCs), irrespective of temperature, showing major declines in all treated turkey samples from days 0–21 of storage. The shelf-lives of untreated, Citrox-treated, and chitosan and Citrox/chitosan-treated samples (as determined by TVC and sensory data) were 13, 17, and >21 days, respectively, at 4 °C for VP turkey. The addition of Citrox was more effective against S . enterica than E . coli in turkey, causing reductions of >0.5 and 2 log cfu/g at 4 and 10 °C, respectively, after 21 days of storage. Interestingly, the addition of chitosan had a significant inhibitory effect on E . coli at 4 °C and S . enterica at 10 °C as compared with the control (inoculated samples) resulting in dramatic reductions in E . coli (2 log) and S . enterica (5 log) cell counts on day 21. Of all the treatments examined, citrus extract in combination with chitosan showed an additive inhibitory effect against both pathogens, reducing E . coli and S . enterica populations, by approximately 2.7 or 4.5 and 2.2 or 5.6 log cfu/g, respectively, at 4 and 10 °C on day 21 of storage. [ABSTRACT FROM AUTHOR]

Phenolic compounds associated with essential oils of spices and herbs possess a variety of antioxidant and antimicrobial properties that interfere with Salmonella detection from fresh and dried products. Finding a compound to neutralize the effect of these antimicrobial compounds, while allowing Salmonella growth during pre-enrichment, is a crucial step in both traditional pathogen isolation and molecular detection from these foods. This study evaluated the effectiveness of corn oil as a component of the pre-enrichment broth to counteract antimicrobial compounds properties and increase the recovery of Salmonella from spices. Oregano samples artificially contaminated with Salmonella enterica were pre-enriched in modified Buffered Peptone Water (mBPW) supplemented with and without 2% (vol/vol) corn oil respectively. Samples were incubated overnight at 37 °C. The results showed that recovery of Salmonella from oregano samples was increased by ≥50% when pre-enriched with corn oil. Serovars were confirmed using a PCR serotyping method. In addition, shot-gun metagenomics analyses demonstrated bacterial diversity and the effect of corn oil on the relative prevalence of Salmonella in the oregano samples. Modifying pre-enrichment broths with corn oil improved the detection and isolation of Salmonella from oregano, and may provide an alternative method for pathogen detection in dried food matrices such as spices. [ABSTRACT FROM AUTHOR]