{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T01:13:39Z","timestamp":1792113219615,"version":"4.4.0"},"reference-count":47,"publisher":"American Society for Microbiology","issue":"8","license":[{"start":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T00:00:00Z","timestamp":1786492800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T00:00:00Z","timestamp":1786492800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["R01AI153605"],"award-info":[{"award-number":["R01AI153605"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["R01AI179807"],"award-info":[{"award-number":["R01AI179807"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["K01AI180357"],"award-info":[{"award-number":["K01AI180357"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000062","name":"National Institute of Diabetes and Digestive and Kidney Diseases","doi-asserted-by":"publisher","award":["P30DK034854"],"award-info":[{"award-number":["P30DK034854"]}],"id":[{"id":"10.13039\/100000062","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100023300","name":"Institut Pasteur","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100023300","id-type":"DOI","asserted-by":"publisher"}]},{"name":"LabEX IBEID"},{"DOI":"10.13039\/100011495","name":"Massachusetts Life Sciences Center","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100011495","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["mBio"],"published-print":{"date-parts":[[2026,8,12]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:sec>\n                    <jats:title\/>\n                    <jats:p>\n                      <jats:italic toggle=\"yes\">Clostridioides difficile<\/jats:italic>\n                      is a toxin-producing pathogen that opportunistically infects those with a depleted gut microbiome, often triggered by antibiotic use.\n                      <jats:italic toggle=\"yes\">C. difficile<\/jats:italic>\n                      preferentially utilizes Stickland amino acids, including proline, to promote energy generation and growth for colonization. We evaluated host outcomes from infection with wild-type and \u0394\n                      <jats:italic toggle=\"yes\">prdB<\/jats:italic>\n                      mouse-infective\n                      <jats:italic toggle=\"yes\">C. difficile<\/jats:italic>\n                      ATCC 43255 strains to investigate how proline metabolism modulates\n                      <jats:italic toggle=\"yes\">C. difficile\u2019s<\/jats:italic>\n                      pathogenesis and interactions with commensals in an enriched gut nutrient environment. While gnotobiotic mice infected with the \u0394\n                      <jats:italic toggle=\"yes\">prdB<\/jats:italic>\n                      mutant succumbed to infection, they showed delayed colonization and toxin production, thereby extending their survival.\n                      <jats:italic toggle=\"yes\">In vivo C. difficile<\/jats:italic>\n                      transcriptomic analyses demonstrated a shift from Stickland fermentation to carbohydrate metabolism in the \u0394\n                      <jats:italic toggle=\"yes\">prdB<\/jats:italic>\n                      mutant. To investigate functions of\n                      <jats:italic toggle=\"yes\">C. difficile\u2019s<\/jats:italic>\n                      proline reductase in interactions with the commensal microbiota, we evaluated infection outcomes in mice co-colonized with the disease-promoting commensal\n                      <jats:italic toggle=\"yes\">Clostridium sardiniense<\/jats:italic>\n                      (CSAR). One-third of CSAR and \u0394\n                      <jats:italic toggle=\"yes\">prdB-<\/jats:italic>\n                      infected mice survived, while co-colonized mice with the wild-type strain rapidly succumbed within 48 h.\n                      <jats:italic toggle=\"yes\">In vivo<\/jats:italic>\n                      transcriptomic analyses from co-colonized mice identified additional metabolic defects in the \u0394\n                      <jats:italic toggle=\"yes\">prdB<\/jats:italic>\n                      mutant, including failure to express oxidative Stickland pathways and ornithine fermentations to harness CSAR-produced ornithine, suggesting conversion of the mutant into a glycolytic strain that was now in direct competition with CSAR for carbohydrate substrates. CSAR remained metabolically active during the \u0394\n                      <jats:italic toggle=\"yes\">prdB<\/jats:italic>\n                      infection through its mucin degradation and polysaccharide and disaccharide metabolism. Our findings illustrate critical functions of\n                      <jats:italic toggle=\"yes\">C. difficile\u2019s<\/jats:italic>\n                      proline reductase in coordinating early metabolism to facilitate gut colonization, including interactions with a cross-feeding commensal species.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE<\/jats:title>\n                    <jats:p>\n                      <jats:italic toggle=\"yes\">Clostridioides difficile<\/jats:italic>\n                      is a spore-forming bacterium that commonly causes pseudomembranous colitis in patients exposed to antibiotics, with infections leading to 30,000 deaths annually in the United States. We conducted further\n                      <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                      and\n                      <jats:italic toggle=\"yes\">in vivo<\/jats:italic>\n                      analyses to determine how proline reductase modulates\n                      <jats:italic toggle=\"yes\">C. difficile<\/jats:italic>\n                      colonization, growth, and metabolism, and showed that this pathway is critical to\n                      <jats:italic toggle=\"yes\">Clostridium sardiniense\u2019<\/jats:italic>\n                      s ability to cross-feed with the pathogen to cause toxic megacolon. We demonstrated that sporulation and toxin production are delayed when\n                      <jats:italic toggle=\"yes\">C. difficile\u2019<\/jats:italic>\n                      s proline reductase pathway is interrupted through the deletion of the\n                      <jats:italic toggle=\"yes\">prdB<\/jats:italic>\n                      gene. Survival was enhanced in mice co-colonized with\n                      <jats:italic toggle=\"yes\">C. sardiniense<\/jats:italic>\n                      and the\n                      <jats:italic toggle=\"yes\">C. difficile<\/jats:italic>\n                      \u0394\n                      <jats:italic toggle=\"yes\">prdB<\/jats:italic>\n                      mutant, as\n                      <jats:italic toggle=\"yes\">C. difficile<\/jats:italic>\n                      relies on glycolytic pathways over Stickland fermentations. The present findings support the central role of proline reductase metabolism in early pathogen growth, metabolism, and toxin, and as a potential therapeutic target against\n                      <jats:italic toggle=\"yes\">C. difficile infection<\/jats:italic>\n                      .\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/mbio.01180-26","type":"journal-article","created":{"date-parts":[[2026,6,22]],"date-time":"2026-06-22T13:02:02Z","timestamp":1782133322000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":1,"title":["Mechanisms by which proline reductase of\n                    <i>Clostridioides difficile<\/i>\n                    promotes efficient metabolism and disease progression\n                    <i>in vivo<\/i>"],"prefix":"10.1128","volume":"17","author":[{"given":"Laura M.","family":"Cersosimo","sequence":"first","affiliation":[{"name":"Massachusetts Host-Microbiome Center, Department Pathology, Brigham & Women's Hospital, Harvard Medical School. 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