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Applied and Environmental Microbiology, December 2005, p. 8362-8370, Vol. 71, No. 12
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.12.8362-8370.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Molecular Characterization of Clostridium perfringens Isolates from Humans with Sporadic Diarrhea: Evidence for Transcriptional Regulation of the Beta2-Toxin-Encoding Gene

Ben Harrison,1 Deepa Raju,1 Helen S. Garmory,2 Moira M. Brett,3 Richard W. Titball,2 and Mahfuzur R. Sarker1*

Department of Microbiology, Oregon State University, Corvallis, Oregon 97331,1 Department of Biomedical Sciences, Defence Science and Technology Laboratory, Porton Down, Salisbury, Wilshire SP4 0JQ, United Kingdom,2 Health Protection Agency and Reference Microbiology Division, London NW9 5HT, United Kingdom3

Received 18 May 2005/ Accepted 6 September 2005

Clostridium perfringens type A food poisoning is caused by C. perfringens isolates carrying a chromosomal enterotoxin gene (cpe), while non-food-borne gastrointestinal (GI) diseases, such as antibiotic-associated diarrhea (AAD) and sporadic diarrhea (SD), are caused by C. perfringens plasmid cpe isolates. A recent study reported the association of beta2 toxin (CPB2) with human GI diseases, and particularly AAD/SD, by demonstrating that a large percentage of AAD/SD isolates, in contrast to a small percentage of food poisoning isolates, carry the beta2-toxin gene (cpb2). This putative relationship was further tested in the current study by characterizing 14 cpe+ C. perfringens fecal isolates associated with recent cases of human SD in England (referred to hereafter as SD isolates). These SD isolates were all classified as cpe+ type A, and 12 of the 14 cpe+ isolates carry their cpe gene on the plasmid and 2 carry it on the chromosome. Interestingly, cpb2 is present in only 12 plasmid cpe isolates; 11 isolates carry cpe and cpb2 on different plasmids, but cpe and cpb2 are located on the same plasmid in one isolate. C. perfringens enterotoxin is produced by all 14 cpe+ SD isolates. However, only 10 of the 12 cpe+/cpb2+ SD isolates produced CPB2, with significant variation in amounts. The levels of cpb2 mRNA in low- to high-CPB2-producing SD isolates differed to such an extent (30-fold) that this difference could be considered a major cause of the differential level of CPB2 production in vitro by SD isolates. Furthermore, no silent or atypical cpb2 was found in a CPB2 Western blot-negative isolate, 5422/94, suggesting that the lack of CPB2 production in 5422/94 was due to low expression of cpb2 mRNA. This received support from our observation that the recombinant plasmid carrying 5422/94 cpb2, which overexpressed cpb2 mRNA, restored CPB2 production in F4969 (a cpb2-negative isolate). Collectively, our present results suggest that CPB2 merits further study as an accessory toxin in C. perfringens-associated SD.

* Corresponding author. Mailing address: Department of Microbiology, Oregon State University, 220 Nash Hall, Corvallis, OR 97331. Phone: (541) 737-2950. Fax: (541) 737-0496. E-mail: sarkerm{at}oregonstate.edu.

Applied and Environmental Microbiology, December 2005, p. 8362-8370, Vol. 71, No. 12
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.12.8362-8370.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Paredes-Sabja, D., Udompijitkul, P., Sarker, M. R. (2009). Inorganic Phosphate and Sodium Ions Are Cogerminants for Spores of Clostridium perfringens Type A Food Poisoning-Related Isolates. Appl. Environ. Microbiol. 75: 6299-6305 [Abstract] [Full Text]  


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