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Shiga toxin-producing Escherichia coli (STEC), a significant cause of foodborne illnesses, is often associated with the consumption of fresh produce, including alfalfa sprouts. This study was executed to determine how quickly STEC grows, adapts, and colonizes alfalfa sprouts during production and storage, and whether the pathogen's virulence and infectious doses are affected by physiological adaptation to sprouts as an environment. A reporter STEC O157:H7 EDL933 strain was developed to track the transcription of eae, a virulence gene involved in colonizing human intestinal enterocytes. When the seeds were inoculated with 2.1 × 10 3  CFU/g of the reporter strain, the pathogen's population increased to 1.5 × 10 6  CFU/g sprouts within 1.38 days and then remained stable during the remainder of the 5-day sprouting, indicating physiological adaptation to this environment. Seeds were inoculated with ∼10 8  CFU/g and subsequently treated with 2000 ppm calcium hypochlorite solution, followed by a water-rinse (treated seeds), or just rinsed with water (untreated seeds). After 5 days of sprouting, the resulting fresh sprouts were refrigerated for three days at 4 °C. Sprout samples were collected and treated with 2000 ppm calcium hypochlorite solution and rinsed thoroughly with water before counting internalized STEC, or just water-washed before measuring total STEC. The transcription of eae (normalized to cell count) was the highest on the second day of sprouting, but the transcription of other virulence and stress-related genes varied, with sodA being upregulated in STEC cells. Lethal dose 50 (LD 50 ) to Galleria mellonella, a STEC infection animal model, was lower (i.e., virulence was higher) in total STEC collected from fresh sprouts produced from treated seeds, compared to that from untreated seeds (1.9 × 10 0 and 6.0 × 10 1  CFU/larva, respectively). Compared to refrigerated sprouts, the LD 50 of STEC from freshly produced sprouts was lower. Based on these findings, it can be concluded that (a) STEC quickly adapts physiologically to sprouts as an environment, (b) transcription of STEC virulence genes changed during sprouts production but generally decreased during refrigeration, and (c) STEC from fresh sprouts grown from sanitizer-treated seeds were more virulent in the animal model, but STEC from refrigerated sprouts were less virulent., 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 © 2024 Elsevier B.V. All rights reserved.)

Bacteriophages have been envisioned as tools to control a variety of foodborne pathogenic bacteria. Salmonella is a foodborne pathogen that is a threat to public health around the world. Contaminated tomatoes have been associated with several Salmonella outbreaks. Hence, the objective of this work was to identify and characterize different lytic bacteriophages against Salmonella Newport, as one of top ten Salmonella serovars associated with human salmonellosis in North America, and then apply these phages to enhance the safety of cherry tomatoes. Four lytic phages against Salmonella Newport were selected based on their ability to lyse a majority of the 26 screened Salmonella serovars. The selected phages belong to Myoviridae (vB_SnwM_CGG4-1, vB_SnwM_CGG4-2) and Siphoviridae (vB_SnwM_CGG3-1, vB_SnwM_CGG3-2) families. They were found to be stable at different temperatures and pH, have latent periods ranging from 53 to 65 min and burst sizes from 92 to 177. In addition, the two Myoviridae phages have a lower frequency of developing bacteriophage insensitive mutants when compared with the Siphoviridae phages. No significant change in virulence gene expression was observed in the developed bacteriophage insensitive mutants when compared to the parental phage sensitive strain. Furthermore, the vB_SnwM_CGG4-1 genome revealed no homology to virulence or lysogenic genes. A phage cocktail was used to control the growth of S. Newport in broth medium and on contaminated cherry tomato. Complete inhibition of bacterial growth in broth medium was observed at 25 °C for 24 h. In addition, a 4.5 log 10 unit reduction in the bacterial count was observed when applying the phage cocktail onto contaminated tomatoes stored at 22 °C for 3 days. These findings suggest that the isolated phages can be used for biocontrol of S. Newport to improve the safety of ready-to-eat (RTE) produce., (Copyright © 2019. Published by Elsevier B.V.)