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Applied and Environmental Microbiology, April 2003, p. 2372-2376, Vol. 69, No. 4
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.4.2372-2376.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Production of Diarrheal Enterotoxins and Other Potential Virulence Factors by Veterinary Isolates of Bacillus Species Associated with Nongastrointestinal Infections

Neil J. Rowan,1* George Caldow,2 Curtis G. Gemmell,3 and Iain S. Hunter1

Department of Bioscience, University of Strathclyde,1 Department of Bacteriology and Immunology, University of Glasgow Medical School, Glasgow Royal Infirmary, Glasgow,3 Veterinary Science Division, Scottish Agricultural College, St. Boswells, Scotland2

Received 11 September 2002/ Accepted 16 January 2003

With the exceptions of Bacillus cereus and Bacillus anthracis, Bacillus species are generally perceived to be inconsequential. However, the relevance of other Bacillus species as food poisoning organisms and etiological agents in nongastrointestinal infections is being increasingly recognized. Eleven Bacillus species isolated from veterinary samples associated with severe nongastrointestinal infections were assessed for the presence and expression of diarrheagenic enterotoxins and other potential virulence factors. PCR studies revealed the presence of DNA sequences encoding hemolysin BL (HBL) enterotoxin complex and B. cereus enterotoxin T (BceT) in five B. cereus strains and in Bacillus coagulans NB11. Enterotoxin HBL was also harbored by Bacillus polymyxa NB6. After 18 h of growth in brain heart infusion broth, all seven Bacillus isolates carrying genes encoding enterotoxin HBL produced this toxin. Cell-free supernatant fluids from all 11 Bacillus isolates demonstrated cytotoxicity toward human HEp-2 cells; only one Bacillus licheniformis strain adhered to this test cell line, and none of the Bacillus isolates were invasive. This study constitutes the first demonstration that Bacillus spp. associated with serious nongastrointestinal infections in animals may harbor and express diarrheagenic enterotoxins traditionally linked to toxigenic B. cereus.

* Corresponding author. Mailing address: Department of Bioscience, University of Strathclyde, Royal College, 204 George St., Glasgow G1 1XW, Scotland. Phone: 44 (0)141 548 2531. Fax: 44 (0)141 553 4124. E-mail: n.j.rowan{at}strath.ac.uk.

Applied and Environmental Microbiology, April 2003, p. 2372-2376, Vol. 69, No. 4
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.4.2372-2376.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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