Epsilon-toxin plasmids of Clostridium perfringens type D are conjugative

Meredith L. Hughes, Rachael Poon, Vicki Adams, Sameera Sayeed, Juliann Saputo, Francisco A Uzal, Bruce A. McClane, Julian I. Rood

Research output: Contribution to journalArticle

51 Citations (Scopus)

Abstract

Isolates of Clostridium perfringens type D produce the potent epsilon-toxin (a CDC/U.S. Department of Agriculture overlap class B select agent) and are responsible for several economically significant enterotoxemias of domestic livestock. It is well established that the epsilon-toxin structural gene, etx, occurs on large Plasmids. We show here that at least two of these plasmids are conjugative. The etx gene on these plasmids was insertionally inactivated using a chloramphenicol resistance cassette to phenotypically tag the plasmid. High-frequency conjugative transfer of the tagged plasmids into the C. perfringens type A strain JIR325 was demonstrated, and the resultant transconjugants were shown to act as donors in subsequent mating experiments. We also demonstrated the transfer of "unmarked" native ε-toxin plasmids into strain JIR325 by exploiting the high transfer frequency. The transconjugants isolated in these experiments expressed functional ε-toxin since their supernatants had cytopathic effects on MDCK cells and were toxic in mice. Using the widely accepted multiplex PCR approach for toxin genotyping, these type A-derived transconjugants were genotypically type D. These findings have significant implications for the C. perfringens typing system since it is based on the toxin profile of each strain. Our study demonstrated the fluid nature of the toxinotypes and their dependence upon the presence or absence of toxin plasmids, some of which have for the first time been shown to be conjugative.

Original languageEnglish (US)
Pages (from-to)7531-7538
Number of pages8
JournalJournal of Bacteriology
Volume189
Issue number21
DOIs
StatePublished - Nov 2007

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Plasmids
Clostridium perfringens
Enterotoxemia
Chloramphenicol Resistance
United States Department of Agriculture
Madin Darby Canine Kidney Cells
Poisons
Multiplex Polymerase Chain Reaction
Livestock
Centers for Disease Control and Prevention (U.S.)
Clostridium perfringens epsilon-toxin
Genes

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Immunology

Cite this

Hughes, M. L., Poon, R., Adams, V., Sayeed, S., Saputo, J., Uzal, F. A., ... Rood, J. I. (2007). Epsilon-toxin plasmids of Clostridium perfringens type D are conjugative. Journal of Bacteriology, 189(21), 7531-7538. https://doi.org/10.1128/JB.00767-07

Epsilon-toxin plasmids of Clostridium perfringens type D are conjugative. / Hughes, Meredith L.; Poon, Rachael; Adams, Vicki; Sayeed, Sameera; Saputo, Juliann; Uzal, Francisco A; McClane, Bruce A.; Rood, Julian I.

In: Journal of Bacteriology, Vol. 189, No. 21, 11.2007, p. 7531-7538.

Research output: Contribution to journalArticle

Hughes, ML, Poon, R, Adams, V, Sayeed, S, Saputo, J, Uzal, FA, McClane, BA & Rood, JI 2007, 'Epsilon-toxin plasmids of Clostridium perfringens type D are conjugative', Journal of Bacteriology, vol. 189, no. 21, pp. 7531-7538. https://doi.org/10.1128/JB.00767-07
Hughes, Meredith L. ; Poon, Rachael ; Adams, Vicki ; Sayeed, Sameera ; Saputo, Juliann ; Uzal, Francisco A ; McClane, Bruce A. ; Rood, Julian I. / Epsilon-toxin plasmids of Clostridium perfringens type D are conjugative. In: Journal of Bacteriology. 2007 ; Vol. 189, No. 21. pp. 7531-7538.
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