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Infection and Immunity, May 2007, p. 2391-2398, Vol. 75, No. 5
0019-9567/07/$08.00+0     doi:10.1128/IAI.02014-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Virulence Plasmid Diversity in Clostridium perfringens Type D Isolates{triangledown} ,{dagger}

Sameera Sayeed, Jihong Li, and Bruce A. McClane*

Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15261

Received 22 December 2006/ Returned for modification 10 February 2007/ Accepted 19 February 2007

Clostridium perfringens type D isolates are important in biodefense and also cause natural enterotoxemias in sheep, goats, and occasionally cattle. In these isolates, the gene (etx) encoding {varepsilon}-toxin is thought to reside on poorly characterized large plasmids. Type D isolates sometimes also produce other potentially plasmid-encoded toxins, including C. perfringens enterotoxin and beta2 toxin, encoded by the cpe and cbp2 genes, respectively. In the current study we demonstrated that the etx, cpe, and cpb2 genes are carried on plasmids in type D isolates and characterized the toxin-encoding plasmids to obtain insight into their genetic organization, potential transferability, and diversity. Southern blotting of pulsed-field gels showed that the etx gene of type D isolates can be present on at least five different plasmids, whose sizes range from 48 to 110 kb. The etx plasmids also typically carried IS1151 and tcp open reading frames (ORFs) known to mediate conjugative transfer of C. perfringens plasmid pCW3. PCR studies revealed that other than their tcp ORFs, etx plasmids of type D isolates do not carry substantial portions of the conserved or variable regions in the cpe plasmids of type A isolates. Southern blotting also demonstrated that in type D isolates the cpe and cpb2 genes are sometimes present on the etx plasmid. Collectively, these findings confirmed that the virulence of type D isolates is heavily plasmid dependent and indicated that (i) a single type D isolate can carry multiple virulence plasmids, (ii) a single type D virulence plasmid can carry up to three different toxin genes, and (iii) many etx plasmids should be capable of conjugative transfer.

* Corresponding author. Mailing address: E1240 BSTWR, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261. Phone: (412) 648-9022. Fax: (412) 624-1401. E-mail: bamcc{at}pitt.edu

{triangledown} Published ahead of print on 5 March 2007.

{dagger} Supplemental material for this article may be found at http://iai.asm.org/.

Editor: D. L. Burns

Infection and Immunity, May 2007, p. 2391-2398, Vol. 75, No. 5
0019-9567/07/$08.00+0     doi:10.1128/IAI.02014-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Miyamoto, K., Li, J., Sayeed, S., Akimoto, S., McClane, B. A. (2008). Sequencing and Diversity Analyses Reveal Extensive Similarities between Some Epsilon-Toxin-Encoding Plasmids and the pCPF5603 Clostridium perfringens Enterotoxin Plasmid. J. Bacteriol. 190: 7178-7188 [Abstract] [Full Text]  
  • Parsons, J. A., Bannam, T. L., Devenish, R. J., Rood, J. I. (2007). TcpA, an FtsK/SpoIIIE Homolog, Is Essential for Transfer of the Conjugative Plasmid pCW3 in Clostridium perfringens. J. Bacteriol. 189: 7782-7790 [Abstract] [Full Text]  
  • Hughes, M. L., Poon, R., Adams, V., Sayeed, S., Saputo, J., Uzal, F. A., McClane, B. A., Rood, J. I. (2007). Epsilon-Toxin Plasmids of Clostridium perfringens Type D Are Conjugative. J. Bacteriol. 189: 7531-7538 [Abstract] [Full Text]  


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