title: Inducing and Quantifying Clostridium difficile Spore Formation.
abstract: The Gram-positive nosocomial pathogen Clostridium difficile induces sporulation during growth in the gastrointestinal tract. Sporulation is necessary for this obligate anaerobe to form metabolically dormant spores that can resist antibiotic treatment, survive exit from the mammalian host, and transmit C. difficile infections. In this chapter, we describe a method for inducing C. difficile sporulation in vitro. This method can be used to study sporulation and maximize spore purification yields for a number of C. difficile strain backgrounds. We also describe procedures for visualizing spore formation using phase-contrast microscopy and for quantifying the efficiency of sporulation using heat resistance as a measure of functional spore formation.
authors: Department of Microbiology and Molecular Genetics, University of Vermont, 95 Carrigan Drive, Burlington, VT, 05405, USA. aimee.shen@uvm.edu.
authors: Aimee
authors: A
authors: Shen
authors: Department of Microbiology and Molecular Genetics, University of Vermont, 95 Carrigan Drive, Burlington, VT, 05405, USA.|Program in Cellular, Molecular & Biomedical Sciences, University of Vermont, Burlington, VT, 05405, USA.
authors: Kelly A
authors: KA
authors: Fimlaid
authors: Department of Microbiology and Molecular Genetics, University of Vermont, 95 Carrigan Drive, Burlington, VT, 05405, USA.
authors: Keyan
authors: K
authors: Pishdadian
mesh_terms: D000693:Anaerobiosis; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D058491:Bacterial Load; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D016360:Clostridioides difficile / Q000187:drug effects / Q000254:growth & development* / Q000648:ultrastructure; D015870:Gene Expression; D006358:Hot Temperature; D008780:Methyltransferases / Q000235:genetics / Q000378:metabolism; D008858:Microscopy, Phase-Contrast; D009154:Mutation; D009962:Orotate Phosphoribosyltransferase / Q000235:genetics / Q000378:metabolism; D013171:Spores, Bacterial / Q000187:drug effects / Q000254:growth & development / Q000302:isolation & purification* / Q000648:ultrastructure; D013656:Taurocholic Acid / Q000494:pharmacology*; D013839:Thiamphenicol / Q000494:pharmacology; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D014157:Transcription Factors; D013656:Taurocholic Acid; D008780:Methyltransferases; C431709:ErmA protein, Bacteria; D009962:Orotate Phosphoribosyltransferase; D013839:Thiamphenicol
keywords: Induction; Phase-contrast microscopy; Spore; Sporulation