Salmonella and Shiga Toxin-Producing Escherichia coli in Products Sampled in the Food Safety and Inspection Service Raw Pork Baseline Study.

The Food Safety and Inspection Service (FSIS) conducts microbiological baseline studies to determine national prevalence of select foodborne pathogens in federally inspected meat and poultry products and to obtain data for risk assessments. The FSIS conducted a baseline study from 1 June 2017 through 31 May 2018 to characterize and determine the prevalence of Salmonella and assess the occurrence of Shiga toxin-producing Escherichia coli (STEC) in a variety of raw pork products. In total, 4,014 samples from slaughter and processing establishments were analyzed for Salmonella; a subset of these samples (1,395) from slaughter establishments were also analyzed for STEC. Analyses determined that the national prevalence of Salmonella in raw pork products was highest in comminuted products (28.9%), followed by intact cuts (5.3%) and nonintact cuts (3.9%). Less than 1% of samples analyzed were positive for the top seven STEC. Our findings indicate there is a need for additional pathogen reduction strategies for raw pork products.

Nanoparticles functionalized with ampicillin destroy multiple-antibiotic-resistant isolates of Pseudomonas aeruginosa and Enterobacter aerogenes and methicillin-resistant Staphylococcus aureus.

We show here that silver nanoparticles (AgNP) were intrinsically antibacterial, whereas gold nanoparticles (AuNP) were antimicrobial only when ampicillin was bound to their surfaces. Both AuNP and AgNP functionalized with ampicillin were effective broad-spectrum bactericides against Gram-negative and Gram-positive bacteria. Most importantly, when AuNP and AgNP were functionalized with ampicillin they became potent bactericidal agents with unique properties that subverted antibiotic resistance mechanisms of multiple-drug-resistant bacteria.

Histone-like nucleoid-structuring protein represses transcription of the ehx operon carried by locus of enterocyte effacement-negative Shiga toxin-expressing Escherichia coli.

Shiga toxin-producing Escherichia coli (STEC) are a significant cause of zoonotic foodborne diarrheal disease in industrialized nations. In addition to Shiga toxin most STEC produce the enterohemolysin (EhxA) toxin. The EhxA toxin is encoded by the ehxCABD operon located on the large plasmid carried by STEC, yet its role in pathogenesis is unknown. A histone-like nucleoid-structuring protein (H-NS) null mutant of STEC O91:H21 strain B2F1 displayed a hyper-hemolytic phenotype, was defective in binding to human colonic epithelial cells, and was non-motile. We concluded that H-NS modulated expression of several genes in B2F1 including the ehx operon. Electrophoretic mobility shift assays indicate that H-NS binds to an 88bp region of DNA upstream of the ehxC start codon. To determine if the same region of DNA was sensitive to repression by H-NS, a transcriptional fusion was constructed between the putative promoter region of ehx and a promoterless lacZ gene. The beta-galactosidase activity detected was low in E. coli that produced H-NS but was significantly higher in the H-NS null background. Taken together, the data indicates that in STEC the 88bp region upstream of the ehx operon contains a cis-acting element to which H-NS binds and negatively regulates expression of enterohemolysin.

Mapping critical interactive sites within the periplasmic domain of the Vibrio cholerae type II secretion protein EpsM.

The type II secretion (T2S) system is present in many gram-negative species, both pathogenic and nonpathogenic, where it supports the delivery of a variety of toxins, proteases, and lipases into the extracellular environment. In Vibrio cholerae, the T2S apparatus is composed of 12 Eps proteins that assemble into a multiprotein complex that spans the entire cell envelope. Two of these proteins, EpsM and EpsL, are key components of the secretion machinery present in the inner membrane. In addition to likely forming homodimers, EpsL and EpsM have been shown to form a stable complex in the inner membrane and to protect each other from proteolytic degradation. To identify and map the specific regions of EpsM involved in protein-protein interactions with both another molecule of EpsM and EpsL, we tested the interactions of deletion constructs of EpsM with full-length EpsM and EpsL by functional characterization and copurification as well as coimmunoprecipitation. Analysis of the truncated EpsM mutants revealed that the region of EpsM from amino acids 100 to 135 is necessary for EpsM to form homo-oligomers, while residues 84 to 99 appear to be critical for a stable interaction with EpsL.

Mutations in hns reduce the adherence of Shiga toxin-producing E. coli 091:H21 strain B2F1 to human colonic epithelial cells and increase the production of hemolysin.

Shiga toxin-producing Escherichia coli (STEC) 091:H21 strain B2F1, an isolate from a patient with the hemolytic uremic syndrome (HUS), produces elastase-activatable Shiga toxin (Stx) type 2d and adheres well to human colonic epithelial T84 cells. This adherence phenotype occurs even though B2F1 does not contain the locus of enterocyte effacement (LEE) that encodes the primary adhesin for E. coli O157:H7. To attempt to identify genes involved in binding of B2F1 to T84 cells a bank of mini-Tn5phoACm(r) transposon mutants of this strain was generated. Several of these mutants exhibited a reduced adherence phenotype, but none of the insertions in these mutants were within putative adhesin genes. Rather, insertional mutations within hns resulted in the loss of adherence. Moreover, the hns mutant also displayed an increase in the production of hemolysin and alkaline phosphatase and a loss of motility with no change in Stx2d-activatable expression levels. When B2F1 was cured of the large plasmid that encodes the hemolysin, the resulting strain adhered well to T84 cells. However, an hns mutant of the plasmid-cured B2F1 strain exhibited a reduction in adherence to T84 cells. Taken together, these results indicate that H-NS regulates the expression of several genes and some potential virulence factors in the intimin-negative B2F1 STEC strain and that the large plasmid is not required for T84 cell colonization.