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Microbiology 154 (2008), 2241-2250; DOI  10.1099/mic.0.2008/016592-0
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Microbiology 154 (2008), 2241-2250; DOI  10.1099/mic.0.2008/016592-0
© 2008 Society for General Microbiology

Germination of spores of Clostridium difficile strains, including isolates from a hospital outbreak of Clostridium difficile-associated disease (CDAD)

Daniel Paredes-Sabja1,{dagger}, Colton Bond1,{dagger}, Robert J. Carman2, Peter Setlow3 and Mahfuzur R. Sarker1,4

1 Department of Biomedical Sciences, Oregon State University, Corvallis, OR 97331, USA
2 TechLab, Inc., 2001 Kraft Drive, Blacksburg, VA 24060, USA
3 Department of Molecular, Microbial and Structural Biology, University of Connecticut Health Center, Farmington, CT 06030, USA
4 Department of Microbiology, Oregon State University, Corvallis, OR 97331, USA

Correspondence
Mahfuzur R. Sarker
sarkerm{at}oregonstate.edu

Clostridium difficile is an emerging nosocomial pathogen and one of the major causes of antibiotic-associated diarrhoea. Cases of Clostridium difficile-associated disease (CDAD) are likely initiated by the ingestion of dormant C. difficile spores, which then germinate, outgrow and rapidly proliferate to cause gastrointestinal (GI) infections. To understand the initial stages of CDAD pathogenesis, we have characterized the germination of spores from a collection of C. difficile strains, including some clinical isolates obtained from a CDAD outbreak (CDAD isolates). Spores of one laboratory strain and five CDAD isolates did not germinate with amino acids, but did germinate on a nutrient-rich medium. However, bile salts had little effect on spore germination, either alone or in a nutrient-rich medium. These spores also germinated with KCl, as well as the non-nutrient germinants dodecylamine and a 1 : 1 chelate of Ca2+ and dipicolinic acid. An unexpected finding was that spores of most of the C. difficile strains also germinated with inorganic phosphate (Pi) with a pH optimum of 6. The in vitro germination of spores of CDAD strains with KCl and Pi, two molecules present at significant levels in the GI tract, suggests that C. difficile spores germinate in the human body by sensing Pi in the early segments of the duodenum and KCl in the colon.


Abbreviations: CDAD, Clostridium difficile-associated disease; CLE, cortex-lytic enzyme; DPA, dipicolinic acid; GI, gastrointestinal; PG, peptidoglycan

{dagger}These authors contributed equally to this work.




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D. Paredes-Sabja, C. Bond, R. J. Carman, P. Setlow, and M. R. Sarker
Germination of spores of Clostridium difficile strains, including isolates from a hospital outbreak of Clostridium difficile-associated disease (CDAD)
Microbiology, April 1, 2009; 155(4): 1376 - 1376.
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