Salmonella enterica ( S. enterica ) is a causative agent of multiple outbreaks of foodborne illness associated with fresh produce, including pre-cut melon and leafy vegetables. Current industrial antimicrobial interventions have been shown to reduce microbial populations by <90%. Consequently, bacteriophages have been suggested as an alternative to chemical sanitizers. Seven S. enterica strains from four serovars (10 5 CFU/mL) were separately inoculated onto excised pieces of Romaine lettuce leaf and cantaloupe flesh treated with a five-strain bacteriophage cocktail 24 h before S. enterica inoculation. S. enterica , total aerobic populations and water activity were measured immediately after inoculation and after 1 and 2 days of incubation at 8 °C. The efficacy of the bacteriophage cocktail varied between strains. Populations of S. enterica Enteritidis strain S3, S. Javiana S203, S. Javiana S200 were reduced by > 3 log CFU/g and S. Newport S2 by 1 log CFU/g on both lettuce and cantaloupe tissues at all sampling times. In contrast, populations of strains S. Thompson S193 and S194 were reduced by 2 log CFU/g on day 0 on lettuce, but were not significantly different (P > 0.05) from the controls thereafter, S. Newport S195 populations were reduced on lettuce by 1 log CFU/g on day 0 and no reductions were found on cantaloupe tissue. Both aerobic populations and water activity were higher on cantaloupe than on lettuce. The water activity of lettuce decreased significantly (P < 0.05) from 0.845 ± 0.027 on day 0–0.494 ± 0.022 on day 1, but that of cantaloupe remained between 0.977 and 0.993 from day 0–2. The results of this study showed that bacteriophages can reduce S. enterica populations on lettuce and cantaloupe tissues but that the magnitude of the effect was strain-dependent.

Multiple outbreaks of foodborne illness caused by Salmonella sp. have been linked to fresh produce. Understanding variability in colonization potential is critical for the selection of experimental strains suitable for research on the ecology of the species in growing food plants. The fate of 43 Salmonella strains from 29 serovars was examined on seedlings from two cultivars of lettuce (‘Winter Density’ and ‘Parris Island Cos’) and tomato (‘Amish Paste’ and ‘Manitoba’). Salmonella populations were measured on xylose lysine deoxycholate agar immediately after inoculation and after 5 days of incubation at 21°C. Laser scanning confocal microscopy was also performed to examine the distribution of cells on seedling leaf surfaces. Laser scanning confocal microscopy showed that cells or cellular aggregates were located within stomata, in surface depressions adjacent to stomata, or on random surface locations on seedlings that were successfully colonized. Populations of 26 strains (60.5%) increased on seedlings from all plant species and cultivars, although there were significant differences (P &lt; 0.05) in the extent of population increase achieved by different strains on the same plant species–cultivar combinations. The remaining strains displayed differential ability to colonize seedlings depending on plant species or cultivar. The results of the present study indicate that the colonization potential of Salmonella is highly variable and should be carefully considered in the selection of experimental strains. HIGHLIGHTS

Salmonella enterica subsp. enterica serovar Enteritidis is able to adapt to sublethal concentrations of ethanol, which subsequently induce tolerance of this pathogen to normally lethal ethanol challenges. This work aims to elucidate the underlying ethanol adaptation mechanisms of S. Enteritidis by proteomic and mutagenic analyses. The global proteomic response of S. Enteritidis to ethanol adaptation (5% ethanol for 1 h) was determined by isobaric tags for relative and absolute quantification (iTRAQ), and it was found that a total of 138 proteins were differentially expressed in ethanol-adapted cells compared to nonadapted cells. A total of 56 upregulated proteins were principally associated with purine metabolism and as transporters for glycine betaine, phosphate, d-alanine, thiamine, and heme, whereas 82 downregulated proteins were mainly involved in enterobactin biosynthesis and uptake, the ribosome, flagellar assembly, and virulence. Moreover, mutagenic analysis further revealed the functions of two highly upregulated proteins belonging to purine metabolism (HiuH, 5-hydroxyisourate hydrolase) and glycine betaine transport (ProX, glycine betaine-binding periplasmic protein) pathways. Deletion of either hiuH or proX resulted in the development of a stronger ethanol tolerance response, suggesting negative regulatory roles in ethanol adaptation. Collectively, this work suggests that S. Enteritidis employs multiple strategies to coordinate ethanol adaptation. IMPORTANCE Stress adaptation in foodborne pathogens has been recognized as a food safety concern since it may compromise currently employed microbial intervention strategies. While adaptation to sublethal levels of ethanol is able to induce ethanol tolerance in foodborne pathogens, the molecular mechanism underlying this phenomenon is poorly characterized. Hence, global proteomic analysis and mutagenic analysis were conducted in the current work to understand the strategies employed by Salmonella enterica subsp. enterica serovar Enteritidis to respond to ethanol adaptation. It was revealed that coordinated regulation of multiple pathways involving metabolism, ABC transporters, regulators, enterobactin biosynthesis and uptake, the ribosome, flagellar assembly, and virulence was responsible for the development of ethanol adaptation response in this pathogen. Such knowledge will undoubtedly contribute to the development and implementation of more-effective food safety interventions.

Novel bacteriophages (phages) possessing a broad host range are consistently and routinely reported, yet there is presently no consensus on the definition of ''broad host range.'' As phages are increasingly being used in the development of methods for the detection and biocontrol of human pathogens, it is important to address the limitations associated with the host range. For instance, unanticipated host range breadth may result in the detection of nonpathogenic targets, thereby increasing the false-positive rate. Moreover, a broad host range is generally favored in biocontrol applications despite the risk of undesirable ancillary effects against nontarget species. Here, we discuss the research progress, applications, and implications of broad host range phages with a focus on tailed broad host range phages infecting human pathogens of concern in the Agri-Food sector.

Nontyphoidal Salmonella enterica is one of the leading pathogens for foodborne outbreaks in a multitude of food commodities, including alfalfa sprouts, which are commonly consumed raw. The food industry has commonly used chlorinated washes, but such methods may not be perceived as natural; this can be a detriment as a large portion of sprouts are designated for the organic market. A natural and affordable antimicrobial method that has been acquiring popularity is the use of bacteriophages. This study compared the efficacy of repeated daily applications and a single application of two separate bacteriophage cocktails (SE14, SE20, SF6 and SE14, SF5, SF6) against four Salmonella enterica (S. enterica) strains on germinating alfalfa sprout seeds from days 0 to 7. The results show S. Enteritidis to be the most susceptible to both cocktails with ~2.5 log CFU/mL decrease on day 0 with cocktail SE14, SF5, and SF6. S. enterica populations on all strains continued to grow even with repeated daily bacteriophage applications but in a significantly decreased rate (p < 0.05) compared with a single bacteriophage application. The extent of the reduction was dependent on the S. enterica strain, but the results do show benefits to using repeated bacteriophage applications during sprout germination to reduce S. enterica populations compared with a single bacteriophage application.

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An emerging consumer trend to purchase minimally heated and ready-to-eat food products may result in processing methods that do not effectively reduce pathogenic populations. Crude Maillard reaction products (MRPs) are naturally generated compounds that have been shown to display antimicrobial effects against pathogens. Crude MRPs were generated from reducing sugars (fructose (Fru), glucose (Glc), ribose (Rib) or xylose (Xyl)) with lysine and the melanoidin equivalence was measured using an absorbance of 420 nm (Ab420). The relative antimicrobial activity of each MRP was measured by examining both the length of lag phase and maximum growth rate. MRPs were found to significantly shorten the lag phase and decrease the maximum growth rate of S. Typhimurium (p < 0.05). Glucose-lysine MRP (GL MRP) was determined to have the highest relative melanoidin (1.690 ± 0.048 at Ab420) and its efficacy against S. Typhimurium populations was measured at 37 °C and at pH 7.0 and estimated on xylose lysine deoxycholate (XLD) agar. GL MRP significantly reduced S. Typhimurium populations by >1 log CFU/mL at 8 and 24 h after inoculation (p < 0.05). GL MRPs also further decreased S. Typhimurium populations significantly under thermal stress condition (55 °C) compared to optimal (37 °C) by ~1 log CFU/mL (p < 0.05). Overall, GL MRP demonstrated effective antimicrobial activity against S. Typhimurium at 37 °C and 55 °C.

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