For all microorganisms, acquisition of metal ions is essential for survival in the environment or in their infected host. Metal ions are required in many biological processes as components of metalloproteins and serve as cofactors or structural elements for enzymes. However, it is critical for bacteria to ensure that metal uptake and availability is in accordance with physiological needs, as an imbalance in bacterial metal homeostasis is deleterious. Indeed, host defense strategies against infection either consist of metal starvation by sequestration or toxicity by the highly concentrated release of metals. To overcome these host strategies, bacteria employ a variety of metal uptake and export systems and finely regulate metal homeostasis by numerous transcriptional regulators, allowing them to adapt to changing environmental conditions. As a consequence, iron, zinc, manganese, and copper uptake systems significantly contribute to the virulence of many pathogenic bacteria. However, during the course of our experiments on the role of iron and manganese transporters in extraintestinal Escherichia coli (ExPEC) virulence, we observed that depending on the strain tested, the importance of tested systems in virulence may be different. This could be due to the different set of systems present in these strains, but literature also suggests that as each pathogen must adapt to the particular microenvironment of its site of infection, the role of each acquisition system in virulence can differ from a particular strain to another. In this review, we present the systems involved in metal transport by Enterobacteria and the main regulators responsible for their controlled expression. We also discuss the relative role of these systems depending on the pathogen and the tissues they infect.

The roles of SitABCD, MntH, and FeoB metal transporters in the virulence of avian pathogenic Escherichia coli (APEC) O78 strain 7122 were assessed using isogenic mutants in chicken infection models. In a single-strain infection model, compared to 7122, the ⌬sit strain demonstrated reduced colonization of the lungs, liver, and spleen. Complementation of the ⌬sit strain restored virulence. In a coinfection model, compared to the virulent APEC strain, the ⌬sit strain demonstrated mean 50-fold, 126-fold, and 25-fold decreases in colonization of the lungs, liver, and spleen, respectively. A ⌬mntH ⌬sit strain was further attenuated, demonstrating reduced persistence in blood and mean 1,400-fold, 954-fold, and 83-fold reduced colonization in the lungs, liver, and spleen, respectively. In coinfections, the ⌬feoB ⌬sit strain demonstrated reduced persistence in blood but increased colonization of the liver. The ⌬mntH, ⌬feoB, and ⌬feoB ⌬mntH strains were as virulent as the wild type in either of the infection models. Strains were also tested for sensitivity to oxidative stress-generating agents. The ⌬mntH ⌬sit strain was the most sensitive strain and was significantly more sensitive than the other strains to hydrogen peroxide, plumbagin, and paraquat. sit sequences were highly associated with APEC and human extraintestinal pathogenic E. coli compared to commensal isolates and diarrheagenic E. coli. Comparative genomic analyses also demonstrated that sit sequences are carried on conjugative plasmids or associated with phage elements and were likely acquired by distinct genetic events among pathogenic E. coli and Shigella sp. strains. Overall, the results demonstrate that SitABCD contributes to virulence and, together with MntH, to increased resistance to oxidative stress.

Background: Avian pathogenic E. coli (APEC) are associated with extraintestinal diseases in poultry. The pstSCAB-phoU operon belongs to the Pho regulon and encodes the phosphate specific transport (Pst) system. A functional Pst system is required for full virulence in APEC and other bacteria and contributes to resistance of APEC to serum, to cationic antimicrobial peptides and acid shock. The global mechanisms contributing to the attenuation and decreased resistance of the APEC pst mutant to environmental stresses have not been investigated at the transcriptional level. To determine the global effect of a pst mutation on gene expression, we compared the transcriptomes of APEC strain χ7122 and its isogenic pst mutant (K3) grown in phosphate-rich medium.

Cheese products are susceptible to postprocessing cross-contamination by bacterial surface contamination during slicing, handling, or packaging, which can lead to food safety issues and significant losses due to spoilage. This study examined the effectiveness of pulsed-light (PL) treatment on the inactivation of the spoilage microorganism Pseudomonas fluorescens, the nonenterohemorrhagic Escherichia coli ATCC 25922 (nonpathogenic surrogate of Escherichia coli O157:H7), and Listeria innocua (nonpathogenic surrogate of Listeria monocytogenes) on cheese surface. The effects of inoculum level and cheese surface topography and the presence of clear polyethylene packaging were evaluated in a full factorial experimental design. The challenge microorganisms were grown to early stationary phase and subsequently diluted to reach initial inoculum levels of either 5 or 7 log cfu/slice. White Cheddar and process cheeses were cut into 2.5×5 cm slices, which were spot-inoculated with 100 µL of bacterial suspension. Inoculated cheese samples were exposed to PL doses of 1.02 to 12.29 J/cm(2). Recovered survivors were enumerated by standard plate counting or the most probable number technique, as appropriate. The PL treatments were performed in triplicate and data were analyzed using a general linear model. Listeria innocua was the least sensitive to PL treatment, with a maximum inactivation level of 3.37±0.2 log, followed by P. fluorescens, with a maximum inactivation of 3.74±0.8 log. Escherichia coli was the most sensitive to PL, with a maximum reduction of 5.41±0.1 log. All PL inactivation curves were nonlinear, and inactivation reached a plateau after 3 pulses (3.07 J/cm(2)). The PL treatments through UV-transparent packaging and without packaging consistently resulted in similar inactivation levels. This study demonstrates that PL has strong potential for decontamination of the cheese surface.

Postprocessing cross-contamination of cheese can lead to both food safety issues and significant losses due to spoilage. Pulsed light (PL) treatment, consisting of short, high-energy, broad-spectrum light pulses, has been proven effective in reducing the microbial load on cheese surface. As PL treatment effectiveness is limited by light-cheese interactions, the possibility to improve its effectiveness by combining it with the antimicrobial nisin was explored. The effect of natamycin, which is added to cheeses as an antifungal agent, on PL effectiveness was also investigated. Pseudomonas fluorescens, Escherichia coli ATCC 25922, and Listeria innocua were used as challenge microorganisms. Bacterial cultures in stationary growth phase were diluted to initial inoculum levels of 5 or 7 log cfu per cheese slice. Slices of sharp white Cheddar cheese and white American singles were cut in rectangles of 2.5 × 5 cm. For cheese slices receiving antimicrobial treatment before PL, slices were dipped in natamycin or nisin, spot inoculated with 100 μL of bacterial suspension, and then treated with PL. Cheese slices receiving PL treatment before antimicrobials were spot inoculated, treated with PL, and then treated with antimicrobials. The PL fluence levels from 1.02 to 12.29 J/cm were used. Survivors were enumerated by standard plate counting or the most probable number technique, as appropriate. All treatments were performed in triplicate, and the data were analyzed using a general linear model. Treatment with nisin or natamycin before PL decreased the effectiveness of PL for all bacteria tested. For instance, PL reduced P. fluorescens on Cheddar cheese by 2.19 ± 0.27 log after 6.14 J/cm, whereas combination treatments at the same PL fluence yielded barely 1 log reduction. Inactivation of L. innocua on Cheddar was only 0.78 ± 0.01 log when using PL after nisin, compared with a 1.30 ± 0.76 log reduction by nisin alone. This was attributed to the absorption of UV light by the 2 antimicrobials, which diminished the UV fluence received by the bacteria. Increased inactivation was obtained when antimicrobials were applied after PL. On process cheese, a maximum reduction of 3.73 ± 0.96 log of L. innocua was obtained at 9.22 J/cm for PL followed by nisin, compared with 3.01 ± 0.48 by PL alone. This study demonstrates that antimicrobials may increase the antimicrobial effectiveness of PL on cheese surface, but the order of treatments is critical.

The objective of this study was to examine the effect of pulsed light (PL) treatment on the color, oxidative stability, and onset of molding of Cheddar cheese. Slices of sharp white Cheddar cheese of 2.5 × 5 cm were treated on one side with PL doses from 1.02 to 12.29 J/cm, sealed in polyethylene bags, and stored at 6°C for up to 1 mo. Peroxide value, color parameters, and the onset of molding were evaluated. No significant changes in color or peroxide value were observed for PL-treated samples compared with the untreated controls. Pulsed light was able to significantly delay surface molding during refrigerated storage, with a PL dose of 9.22 J/cm delaying the onset of molding by 7 d. The effect of PL on the taste, appearance, and acceptability of Cheddar cheese slices treated with a PL dose of 9.22 J/cm on each side was assessed. In triangle tests, 60 untrained panelists were unable to detect significant differences between the control and PL-treated samples, although PL had a significant effect on overall liking, flavor, and appearance. These findings suggest that although PL can be effective for surface decontamination of cheese, it may have some detrimental effects on sensory properties.