This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Häggblom, M. M
Right arrow Articles by Salkinoja-Salonen, M. S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Häggblom, M. M
Right arrow Articles by Salkinoja-Salonen, M. S.
Agricola
Right arrow Articles by Häggblom, M. M
Right arrow Articles by Salkinoja-Salonen, M. S.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, May 2002, p. 2479-2483, Vol. 68, No. 5
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.5.2479-2483.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Quantitative Analysis of Cereulide, the Emetic Toxin of Bacillus cereus, Produced under Various Conditions

Max M Häggblom,1,2* Camelia Apetroaie,1 Maria A. Andersson,1 and Mirja S. Salkinoja-Salonen1

Department of Applied Chemistry and Microbiology, University of Helsinki, FIN-00014 University of Helsinki, Finland,1 Department of Biochemistry and Microbiology and Biotechnology Center for Agriculture and the Environment, Cook College, Rutgers, The State University of New Jersey, New Brunswick, New Jersey 08901-85252

Received 14 September 2001/ Accepted 27 February 2002

This paper describes a quantitative and sensitive chemical assay for cereulide, the heat-stable emetic toxin produced by Bacillus cereus. The methods previously available for measuring cereulide are bioassays that give a toxicity titer, but not an accurate concentration. The dose of cereulide causing illness in humans is therefore not known, and thus safety limits for cereulide cannot be indicated. We developed a quantitative and sensitive chemical assay for cereulide based on high-performance liquid chromatography (HPLC) connected to ion trap mass spectrometry. This chemical assay and a bioassay based on boar sperm motility inhibition were calibrated with purified cereulide and with valinomycin, a structurally similar cyclic depsipeptide. The boar spermatozoan motility assay and chemical assay gave uniform results over a wide range of cereulide concentrations, ranging from 0.02 to 230 µg ml-1. The detection limit for cereulide and valinomycin by HPLC-mass spectrometry was 10 pg per injection. The combined chemical and biological assays were used to define conditions and concentrations of cereulide formation by B. cereus strains F4810/72, NC7401, and F5881. Cereulide production commenced at the end of logarithmic growth, but was independent of sporulation. Production of cereulide was enhanced by incubation with shaking compared to static conditions. The three emetic B. cereus strains accumulated 80 to 166 µg of cereulide g-1 (wet weight) when grown on solid medium. Strain NC7401 accumulated up to 25 µg of cereulide ml-1 in liquid medium at room temperature (21 ± 1°C) in 1 to 3 days, during the stationary growth phase when cell density was 2 x 108 to 6 x 108 CFU ml-1. Cereulide production at temperatures at and below 8°C or at 40°C was minimal.

* Corresponding author. Mailing address: Department of Biochemistry and Microbiology, and Biotechnology Center for Agriculture and the Environment, Cook College, Rutgers, The State University of New Jersey, 76 Lipman Dr., New Brunswick, NJ 08901-8525. Phone: (732) 932-9763, ext. 326. Fax: (732) 932-8965. E-mail: haggblom{at}aesop.rutgers.edu.

Applied and Environmental Microbiology, May 2002, p. 2479-2483, Vol. 68, No. 5
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.5.2479-2483.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Mols, M., Abee, T. (2008). Role of Ureolytic Activity in Bacillus cereus Nitrogen Metabolism and Acid Survival. Appl. Environ. Microbiol. 74: 2370-2378 [Abstract] [Full Text]  
  • Fricker, M., Messelhausser, U., Busch, U., Scherer, S., Ehling-Schulz, M. (2007). Diagnostic Real-Time PCR Assays for the Detection of Emetic Bacillus cereus Strains in Foods and Recent Food-Borne Outbreaks. Appl. Environ. Microbiol. 73: 1892-1898 [Abstract] [Full Text]  
  • Thorsen, L., Hansen, B. M., Nielsen, K. F., Hendriksen, N. B., Phipps, R. K., Budde, B. B. (2006). Characterization of Emetic Bacillus weihenstephanensis, a New Cereulide-Producing Bacterium.. Appl. Environ. Microbiol. 72: 5118-5121 [Abstract] [Full Text]  
  • Gray, K. M., Banada, P. P., O'Neal, E., Bhunia, A. K. (2005). Rapid Ped-2E9 Cell-Based Cytotoxicity Analysis and Genotyping of Bacillus Species. J. Clin. Microbiol. 43: 5865-5872 [Abstract] [Full Text]  
  • From, C., Pukall, R., Schumann, P., Hormazabal, V., Granum, P. E. (2005). Toxin-Producing Ability among Bacillus spp. Outside the Bacillus cereus Group. Appl. Environ. Microbiol. 71: 1178-1183 [Abstract] [Full Text]  
  • Ehling-Schulz, M., Vukov, N., Schulz, A., Shaheen, R., Andersson, M., Martlbauer, E., Scherer, S. (2005). Identification and Partial Characterization of the Nonribosomal Peptide Synthetase Gene Responsible for Cereulide Production in Emetic Bacillus cereus. Appl. Environ. Microbiol. 71: 105-113 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»