YM155 Inhibits NleB and SseK Arginine Glycosyltransferase Activity

The type III secretion system effector proteins NleB and SseK are glycosyltransferases that glycosylate protein substrates on arginine residues. We conducted high-throughput screening assays on 42,498 compounds to identify NleB/SseK inhibitors. Such small molecules may be useful as mechanistic probes and may have utility in the eventual development of anti-virulence therapies against enteric bacterial pathogens. We observed that YM155 (sepantronium bromide) inhibits the activity of Escherichia coli NleB1, Citrobacter rodentium NleB, and both Salmonella enterica SseK1 and SseK2. YM155 was not toxic to mammalian cells, nor did it show cross-reactivity with the mammalian O-linked N-acetylglucosaminyltransferase (OGT). YM155 reduced Salmonella survival in mouse macrophage-like cells but had no direct impact on bacterial growth rates, suggesting YM155 may have utility as a potential anti-virulence inhibitor.

Survival of Salmonella during Apple Dehydration as Affected by Apple Cultivar and Antimicrobial Pretreatment

ABSTRACT Dehydrated fruits, including dried coconut (Cocos nucifera) and dried apple (Malus sp.) slices, have been the subject of manufacturer recalls due to contamination with Salmonella. A study was conducted to determine the survival of Salmonella on apple slices of six apple cultivars after dehydration and also following treatment with antimicrobial solutions (0.5%, w/w) and dehydration. Samples of six apple cultivars (Envy, Gala, Red Delicious, Fuji, Pink Lady, Granny Smith) were cored and sliced into 0.4-cm rings, halved, inoculated with a five-strain composite of desiccation-resistant Salmonella, and dehydrated at 60°C for 5 h. Subsequently, Gala apple slices were treated in 0.5% solutions of one of eight antimicrobial rinses for 2 min and then dehydrated at 60°C for 5 h. Antimicrobial solutions used were potassium sorbate, sodium benzoate, ascorbic acid, propionic acid, lactic acid, citric acid, fumaric acid, and sodium bisulfate. Reduction of Salmonella populations varied according to apple cultivar. Salmonella survival on Envy, Gala, Red Delicious, Fuji, Pink Lady, and Granny Smith was 5.92, 5.58, 4.83, 4.68, 4.45, and 3.84 log CFU, respectively. There was significantly greater (P < 0.05) Salmonella inactivation on Granny Smith, Pink Lady, and Fuji apples than on Gala and Envy. Survival of Salmonella on Gala apple slices following dehydration was 5.58 log CFU for the untreated control and 4.76, 3.90, 3.29, 3.13, 2.89, 2.83, 2.64, and 0.0 log CFU for those treated with potassium sorbate, sodium benzoate, ascorbic acid, propionic acid, lactic acid, citric acid, fumaric acid, and sodium bisulfate, respectively. Pretreatment of apple slices with either fumaric acid or sodium bisulfate before dehydration led to lower Salmonella survival than pretreatment with all other antimicrobial treatments. Lower apple pH was statistically correlated (P < 0.05) with decreasing survival of Salmonella following dehydration. These results may provide methodology applicable to the food industry for increasing the inactivation of Salmonella during the dehydration of apple slices. HIGHLIGHTS

Salmonella Survival in Soil and Transfer Onto Produce via Splash Events

ABSTRACT Nearly one-half of foodborne illnesses in the United States can be attributed to fresh produce consumption. The preharvest stage of production presents a critical opportunity to prevent produce contamination in the field from contaminating postharvest operations and exposing consumers to foodborne pathogens. One produce-contamination route that is not often explored is the transfer of pathogens in the soil to edible portions of crops via splash water. We report here on the results from multiple field and microcosm experiments examining the potential for Salmonella contamination of produce crops via splash water, and the effect of soil moisture content on Salmonella survival in soil and concentration in splash water. In field and microcosm experiments, we detected Salmonella for up to 8 to 10 days after inoculation in soil and on produce. Salmonella and suspended solids were detected in splash water at heights of up to 80 cm from the soil surface. Soil-moisture conditions before the splash event influenced the detection of Salmonella on crops after the splash events—Salmonella concentrations on produce after rainfall were significantly higher in wet plots than in dry plots (geometric mean difference = 0.43 CFU/g; P = 0.03). Similarly, concentrations of Salmonella in splash water in wet plots trended higher than concentrations from dry plots (geometric mean difference = 0.67 CFU/100 mL; P = 0.04). These results indicate that splash transfer of Salmonella from soil onto crops can occur and that antecedent soil-moisture content may mediate the efficiency of microbial transfer. Splash transfer of Salmonella may, therefore, pose a hazard to produce safety. The potential for the risk of splash should be further explored in agricultural regions in which Salmonella and other pathogens are present in soil. These results will help inform the assessment of produce safety risk and the development of management practices for the mitigation of produce contamination. HIGHLIGHTS

Effects of different moisture and temperature levels on Salmonella survival in poultry fat

Fat products have been historically thought to have too low water activity to harbor pathogens. However, it has been recently reported that high moisture levels in fats may lead to Salmonella presence and growth. Limited research on strategies to eliminate pathogens in these environments is available, and the mechanisms contributing to microbial presence and growth are not yet well understood. The purpose of this research was to evaluate the effects of moisture levels and storage temperatures on the growth and survival of Salmonella in poultry fat. Samples were stored for 7 d at 48°C or 76°C and remaining Salmonella was evaluated. When poultry fat was challenged with a wet high inoculum, more than a 4 log CFU/mL difference in Salmonella population was observed with 1% and 3% moisture levels at 48°C after 5 d (P < 0.05). No differences between moisture levels (P > 0.05) were observed when samples were tested with a wet low inoculum. Counts below detectable limits were observed after 24 h in samples challenged at 76°C, regardless of inoculum level. When poultry fat was stored at 48°C and inoculated with low levels of Salmonella spp., bacterial growth was influenced only by time (P < 0.05) and not affected (P > 0.05) by moisture level. However, when poultry fat was stored at 48°C and inoculated with high levels of Salmonella spp., bacterial decrease was easier (P < 0.05) in samples containing greater moisture. This research suggests that residual moisture in containers during transportation of poultry fat largely does not affect Salmonella spp. growth.