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Applied and Environmental Microbiology, November 2007, p. 7358-7366, Vol. 73, No. 22
0099-2240/07/$08.00+0     doi:10.1128/AEM.00582-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Morphological and Genetic Diversity of Temperate Phages in Clostridium difficile{triangledown}

Louis-Charles Fortier1,2* and Sylvain Moineau2,3

Département de Microbiologie et d'Infectiologie, Faculté de Médecine et des Sciences de la Santé, Université de Sherbrooke, Québec, Canada J1H 5N4,1 Département de Biochimie et de Microbiologie, Faculté des Sciences et de Génie, Groupe de Recherche en Écologie Buccale, Faculté de Médecine Dentaire,2 Félix d'Hérelle Reference Centre for Bacterial Viruses, Université Laval, Québec, Canada G1K 7P43

Received 13 March 2007/ Accepted 8 September 2007

Eight temperate phages were characterized after mitomycin C induction of six Clostridium difficile isolates corresponding to six distinct PCR ribotypes. The hypervirulent C. difficile strain responsible for a multi-institutional outbreak (NAP1/027 or QCD-32g58) was among these prophage-containing strains. Observation of the crude lysates by transmission electron microscopy (TEM) revealed the presence of three phages with isometric capsids and long contractile tails (Myoviridae family), as well as five phages with long noncontractile tails (Siphoviridae family). TEM analyses also revealed the presence of a significant number of phage tail-like particles in all the lysates. Southern hybridization experiments with restricted prophage DNA showed that C. difficile phages belonging to the family Myoviridae are highly similar and most likely related to previously described prophages {phi}C2, {phi}C5, and {phi}CD119. On the other hand, members of the Siphoviridae phage family are more genetically divergent, suggesting that they originated from distantly related ancestors. Our data thus suggest that there are at least three genetically distinct groups of temperate phages in C. difficile; one group is composed of highly related myophages, and the other two groups are composed of more genetically heterogeneous siphophages. Finally, no gene homologous to genes encoding C. difficile toxins or toxin regulators could be identified in the genomes of these phages using DNA hybridization. Interestingly, each unique phage restriction profile correlated with a specific C. difficile PCR ribotype.

* Corresponding author. Mailing address: Département de microbiologie et d'infectiologie, Faculté de médecine et des sciences de la santé, Université de Sherbrooke, 3001 12e Ave. Nord, Sherbrooke, Québec, Canada J1H 5N4. Phone: (819) 820-6868, ext. 12785. Fax: (819) 564-5392. E-mail: Louis-Charles.Fortier{at}USherbrooke.ca

{triangledown} Published ahead of print on 21 September 2007.

Applied and Environmental Microbiology, November 2007, p. 7358-7366, Vol. 73, No. 22
0099-2240/07/$08.00+0     doi:10.1128/AEM.00582-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Mayer, M. J., Narbad, A., Gasson, M. J. (2008). Molecular Characterization of a Clostridium difficile Bacteriophage and Its Cloned Biologically Active Endolysin. J. Bacteriol. 190: 6734-6740 [Abstract] [Full Text]  


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