Project description:Clostridioides difficile infection (CDI), caused by strains producing toxin B (TcdB), poses a significant global health threat. While C. difficile exhibits substantial diversity, functional studies have focused on a limited number of isolates, overlooking other genomospecies within the genus. We describe five isolates from patients suspected of having CDI who tested negative for the PaLoc marker gene tcdC. Through genomic, proteomic, and phenotypic analyses, we demonstrate that they correspond to three novel toxin-producing species, designated as Clostridioides cryptodifficilis sp. nov., Clostridioides divergens sp. nov., and Clostridioides subdifficilis sp. nov. These species are distinguished by unique MALDI-ToF signatures, metabolic capabilities, and genomic and proteomic architectures, underscoring their clear taxonomic and functional divergence from C. difficile. They secrete functionally active TcdB7 or TcdB11, as demonstrated by cytotoxicity assays in cultured cells and in vivo using the mouse ileal loop model, implicating them in disease pathology, albeit with lower virulence than C. difficile. Our findings expand the known diversity of TcdB-producing Clostridioides and have direct implications for diagnostics, surveillance, and clinical management of diarrheal diseases.
Project description:Clostridioides difficile infection (CDI), caused by strains producing toxin B (TcdB), poses a significant global health threat. While C. difficile exhibits substantial diversity, functional studies have focused on a limited number of isolates, overlooking other genomospecies within the genus. We describe five isolates from patients suspected of having CDI who tested negative for the PaLoc marker gene tcdC. Through genomic, proteomic, and phenotypic analyses, we demonstrate that they correspond to three novel toxin-producing species, designated as Clostridioides cryptodifficilis sp. nov., Clostridioides divergens sp. nov., and Clostridioides subdifficilis sp. nov. These species are distinguished by unique MALDI-ToF signatures, metabolic capabilities, and genomic and proteomic architectures, underscoring their clear taxonomic and functional divergence from C. difficile. They secrete functionally active TcdB7 or TcdB11, as demonstrated by cytotoxicity assays in cultured cells and in vivo using the mouse ileal loop model, implicating them in disease pathology, albeit with lower virulence than C. difficile. Our findings expand the known diversity of TcdB-producing Clostridioides and have direct implications for diagnostics, surveillance, and clinical management of diarrheal diseases.
2026-05-06 | PXD067952 | Pride
Project description:Clostridioides difficile genomes from clinical isolates
Project description:We used RNA-seq to conduct a genome-wide transcriptomic analysis of the C. difficile strain R20291 carrying the newly introduced ϕCD38-2 prophage. A total of 39 bacterial genes were differentially expressed in the R20291 lysogen, 26 of them being downregulated. Several of the regulated genes encode transcriptional regulators and PTS subunits involved in glucose, fructose and glucitol/sorbitol uptake and metabolism. Also of note, the presence of ϕCD38-2 upregulated the expression of a group of regulatory genes located in phi-027, a resident prophage common to most ribotype 027 isolates. The most differentially expressed gene was the conserved phase-variable cell wall protein CwpV, which was upregulated by about 20-fold in the lysogen. This is the first study describing the global response of C. difficile to the presence of a newly introduced prophage.
2014-12-22 | GSE56818 | GEO
Project description:Sequencing data of Clostridioides difficile clinical isolates
Project description:We used RNA-seq to conduct a genome-wide transcriptomic analysis of the C. difficile strain R20291 carrying the newly introduced M-OM-^UCD38-2 prophage. A total of 39 bacterial genes were differentially expressed in the R20291 lysogen, 26 of them being downregulated. Several of the regulated genes encode transcriptional regulators and PTS subunits involved in glucose, fructose and glucitol/sorbitol uptake and metabolism. Also of note, the presence of M-OM-^UCD38-2 upregulated the expression of a group of regulatory genes located in phi-027, a resident prophage common to most ribotype 027 isolates. The most differentially expressed gene was the conserved phase-variable cell wall protein CwpV, which was upregulated by about 20-fold in the lysogen. This is the first study describing the global response of C. difficile to the presence of a newly introduced prophage. Two strains (WT and lysogen) were subjected for RNA-seq analysis with two biological replicates each.
Project description:Clostridioides difficile (formerly Clostridium difficile) is a Gram-positive, spore-forming pathogen which cases drug-induced Clostridioides difficile-associated diseases in hospitals worldwide. A detailed analysis of the proteome may provide new targets for drug development or therapy strategies to combat this pathogen. So far, quantitative proteome analyses could only be carried out by label-free or chemical labeling methods. However, the application of metabolic labeling would allow for accurate quantification of significant differences, even in the case of very small changes. Additionally, it would be possible to perform bias free studies of the membrane or surface proteome which require elaborated preparations and are therefore prone to higher standard deviations during the quantification. Up to now, the implementation of metabolic labeling strategies of C. difficile was hampered by the very specific metabolic requirements of this anaerobic pathogen. To solve this problem, media were evaluated and the cultivation procedure with 15N labeled media for the C. difficile 630Δerm strain was optimized to gain a high incorporation rate. In the following proof-of-principle experiment, the cytosolic sub-proteomes of C. difficile cells of three different cultivation media and two growth phases were analyzed resulting in reproducible data which are shown in detail.
Project description:Gene expression level of Clostridioides difficile (C. difficile) strain R20291 comparing control C. difficile carring pMTL84151 as vector plasmid with C. difficile conjugated with a pMTL84151-03890 gene. Goal was to determine the effects of 03890 gene conjugation on C. difficile strain R20291 gene expression.