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Bile salt hydrolase (BSH) activity against the bile acid tauro-beta-muricholic acid (T-β-MCA) was recently reported to mediate host bile acid, glucose, and lipid homeostasis via the farnesoid X receptor (FXR) signaling pathway. An earlier study correlated decreased Lactobacillus abundance in the cecum with increased concentrations of intestinal T-β-MCA, an FXR antagonist. While several studies have characterized BSHs in lactobacilli, deconjugation of T-β-MCA remains poorly characterized among members of this genus, and therefore it was unclear what strain(s) were responsible for this activity. Here, a strain of L. johnsonii with robust BSH activity against T-β-MCA in vitro was isolated from the cecum of a C57BL/6J mouse. A screening assay performed on a collection of 14 Lactobacillus strains from nine different species identified BSH substrate specificity for T-β-MCA only in two of three L. johnsonii strains. Genomic analysis of the two strains with this BSH activity revealed the presence of three bsh genes that are homologous to bsh genes in the previously sequenced human-associated strain L. johnsonii NCC533. Heterologous expression of several bsh genes in E. coli followed by enzymatic assays revealed broad differences in substrate specificity even among closely related bsh homologs, and suggests that the phylogeny of these enzymes does not closely correlate with substrate specificity. Predictive modeling allowed us to propose a potential mechanism driving differences in BSH activity for T-β-MCA in these homologs. Our data suggests that L. johnsonii regulates T-β-MCA levels in the mouse intestinal environment, and that this species may play a central role in FXR signaling in the mouse. [ABSTRACT FROM AUTHOR]

Background: Shiga toxin-producing Escherichia coli O157:H7 is a foodborne pathogen that causes severe human diseases including hemolytic uremic syndrome (HUS). The virulence factor that mediates HUS, Shiga toxin (Stx), is encoded within the genome of a lambdoid prophage. Although draft sequences are publicly available for a large number of E. coli O157:H7 strains, the high sequence similarity of stx-converting bacteriophages with other lambdoid prophages poses challenges to accurately assess the organization and plasticity among stx-converting phages due to assembly difficulties. Methods: To further explore genome plasticity of stx-converting prophages, we enriched phage DNA from 45 ciprofloxacin-induced cultures for subsequent 454 pyrosequencing to facilitate assembly of the complete phage genomes. In total, 22 stx2a-converting phage genomes were closed. Results: Comparison of the genomes distinguished nine distinct phage sequence types (PSTs) delineated by variation in obtained sequences, such as single nucleotide polymorphisms (SNPs) and insertion sequence element prevalence and location. These nine PSTs formed three distinct clusters, designated as PST1, PST2 and PST3. The PST2 cluster, identified in two clade 8 strains, was related to stx2a-converting phages previously identified in non- O157 Shiga-toxin producing E. coli (STEC) strains associated with a high incidence of HUS. The PST1 cluster contained phages related to those from E. coli O157:H7 strain Sakai (lineage I, clade 1), and PST3 contained a single phage that was distinct from the rest but most related to the phage from E. coli O157:H7 strain EC4115 (lineage I/II, clade 8). Five strains carried identical stx2a-converting phages (PST1-1) integrated at the same chromosomal locus, but these strains produced different levels of Stx2. Conclusion: The stx2a-converting phages of E. coli O157:H7 can be categorized into at least three phage types. Diversification within a phage type is mainly driven by IS629 and by a small number of SNPs. Polymorphisms between phage genomes may help explain differences in Stx2a production between strains, however our data indicates that genes encoded external to the phage affect toxin production as well. [ABSTRACT FROM AUTHOR]

Background: Escherichia coli can experience a multifaceted life, in some cases acting as a commensal while in other cases causing intestinal and/or extraintestinal disease. Several studies suggest enteroaggregative E. coli are the predominant cause of E. coli-mediated diarrhea in the developed world and are second only to Campylobacter sp. as a cause of bacterialmediated diarrhea. Furthermore, enteroaggregative E. coli are a predominant cause of persistent diarrhea in the developing world where infection has been associated with malnourishment and growth retardation. Methods: In this study we determined the complete genomic sequence of E. coli 042, the prototypical member of the enteroaggregative E. coli, which has been shown to cause disease in volunteer studies. We performed genomic and phylogenetic comparisons with other E. coli strains revealing previously uncharacterised virulence factors including a variety of secreted proteins and a capsular polysaccharide biosynthetic locus. In addition, by using Biolog™ Phenotype Microarrays we have provided a full metabolic profiling of E. coli 042 and the non-pathogenic lab strain E. coli K-12. We have highlighted the genetic basis for many of the metabolic differences between E. coli 042 and E. coli K-12. Conclusion: This study provides a genetic context for the vast amount of experimental and epidemiological data published thus far and provides a template for future diagnostic and intervention strategies. [ABSTRACT FROM AUTHOR]

Salmonella Typhimurium is the leading cause of foodborne illnesses in the U.S., causing over a million cases each year. In recent years, whole-genome sequencing (WGS) has become a standard tool for routine epidemiological subtyping. The objectives of this study are 1) to compare the phenotypic and genotypic antimicrobial resistance (AMR) profiles of multidrug resistant (MDR) S. Typhimurium isolates, 2) to examine the genetic relatedness of a historic collection of MDR and pan-susceptible isolates from retail chickens. We used data on Salmonella Typhimurium isolates in the publicly available NARMS national clinical and retail meat datasets from 2016 to 2018. Staramr (0.5.1) was used to identify AMR determinants and predictive resistance from genomes submitted to NCBI. Sensitivity and specificity of the WGS method were calculated with phenotypic resistance results as the reference. SNP-based cluster analysis was used to examine the genetic relatedness of MDR resistant and pan-susceptible isolates from retail chickens. The overall sensitivity of WGS as a predictor of clinical resistance was 96.47% and the overall specificity was 100.00%. The disagreement between phenotypic and genotypic results were mostly related to streptomycin. The MDR isolates differed by an average of 73.1 SNPs, while the pan-susceptible isolates differed by an average of 473.1 SNPs (p < 0.0001). The nearest distance between a pan-susceptible and an MDR isolate was 547 SNPs. WGS can reliably predict AMR in S. Typhimurium isolates and it can reveal genetic determinants to elucidate the evolution of antimicrobial resistance. • Whole genome sequencing (WGS) can reliably predict AMR in S. Typhimurium isolates. • WGS can reveal genetic determinants to elucidate the evolution of antimicrobial resistance. • The overall specificity of WGS in predicting antimicrobial resistance is 100%. • Multi-drug resistant S. Typhimurium isolates were genetically closer compared to pan-susceptible isolates. [ABSTRACT FROM AUTHOR]