SpringerProtocols: Abstract: 2. Detection and Purification of Bacillus cereus Enterotoxins

2. Detection and Purification of Bacillus cereus Enterotoxins

By: Toril Lindbäck², Per Einar Granum²

Abstract

Full Text

Download PDF (122K)

Bacillus cereus causes two types of food poisoning, emetic and diarrheal. The emetic disease is caused by a small cyclic polypeptide (cereulide), and the diarrheal disease is caused by three different enterotoxins. Commercially available kits are used for detection of two of the enterotoxins. The enterotoxins are secreted by B. cereus in the early stationary phase and can be purified from the growth medium by chromatographic methods. The enterotoxins are membrane-active and the toxicity is tested on Vero cells, while the presence of the emetic toxin is detected using boar spermatozoa. Methods for detection and purification of enterotoxins are described, in addition to detection of the emetic toxin.

Affiliation(s): (2) Department of Food Safety and Infection Biology, The Norwegian School of Veterinary Science, Oslo, Norway

Book Title: Food-Borne Pathogens: Methods and Protocols

Series: Methods in Biotechnology | Volume: 21 | Pub. Date: Oct-06-2005 | Page Range: 15-26 | DOI: 10.1385/1-59259-990-7:015

Subject: Microbiology

Key Words: Bacillus cereus - enterotoxins - emetic toxins - food poisoning

References

Download References

1.	Priest, F. G., Kaji, D. A., Rosato, Y. B., and Canhos, V. P. (1994) Characterization of Bacillus thuringiensis and related bacteria by ribosomal RNA gene restriction fragment length polymorphisms. Microbiology 140, 1015–1022.

2.	Lechner, S., Mayr, R., Francis, K. P., et al. (1998) Bacillus weihenstephanensis sp. nov. is a new psychrotolerant species of the Bacillus cereus group. Int. J. Syst. Bacteriol. 48, 1373–1382.

3.	Nakamura, L. K. (1998) Bacillus pseudomycoides sp. nov. Int. J. Syst. Bacteriol. 48, 1031–1035.

4.	Hauge, S. (1955) Food poisoning caused by aerobic spore-forming bacilli. J. Appl. Bacteriol. 18, 591–595.

5.	Kramer, J. M. and Gilbert, R. J. (1989) Bacillus cereus and other Bacillus species, in Foodborne Bacterial Pathogens (Doyle, M. P., ed.), Marcel Dekker, New York, pp. 21–70.

6.	Granum, P. E. (2001) Bacillus cereus, in Food Microbiology. Fundamentals and Frontiers (Doyle, M., Beuchat, L., and Montville, T., eds.), ASM Press, Washington DC, pp. 373–381.

7.	Logan, N. A. and Berkeley, R. C. (1984) Identification of Bacillus strains using the API system. J. Gen. Microbiol. 130, 1871–1882.

8.	Carlson, C. R., Caugant, D., and Kolstø, A.-B. (1994) Genotypic diversity among Bacillus cereus and Bacillus thuringiensis strains. Appl. Environ. Microbiol. 60, 1719–1725.

9.	Helgason, E., Økstad, O. A., Caugant, D. A., et al. (2000) Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis—one species on the basis of genetic evidence. Appl. Environ. Microbiol. 66, 2627–2630.

10.	Ticknor, L. O., Kolstø, A.-B., Hill, K. K., et al. (2001) Fluorescent amplified fragment length polymorphism analysis of Norwegian Bacillus cereus and Bacillus thuringiensis soil isolates. Appl. Environ. Microbiol. 67, 4863–4873.

11.	Sneath, P. H. A. (1986) Endospore-forming Gram-positive rods and cocci, in Bergey’s Manual of Systematic Bacteriology (Sneath, P. H. A., Mair, N. S., Sharpe, M. E., and Holt, J. G., eds.), Williams and Wilkins, Baltimore, pp. 1104–1207.

12.	Claus, D. and Berkeley, R. C. W. (1986) Genus Bacillus, in Bergey’s Manual of Systematic Bacteriology (Sneath, P. H. A., Mair, N. S., Sharpe, M. E., and Holt, J. G. eds.), Williams and Wilkins, Baltimore, pp. 1105–1138.

13.	Madigan, M. T., Martinko, J. M., and Parker, J. (2003) Enterotoxins, in Brock Biology of Microorganisms (Truehart, C., ed.), Prentice Hall, NJ, pp. 746–748.

14.	Beecher, D. J. and MacMillan, J. D. (1991) Characterization of the components of hemolysin BL from Bacillus cereus. Infect. Immunol. 59, 1778–1784.

15.	Beecher, D. J. and Wong, A. C. (1994) Improved purification and characterization of hemolysin BL, a haemolytic dermonecrotic vascular permeability factor from Bacillus cereus. Infect. Immunol. 62, 980–986.

16.	Beecher, D. J. and Wong, A. C. (1997) Tripartite hemolysin BL from Bacillus cereus. Hemolytic analysis of component interactions and a model for its characteristic paradoxical zone phenomenon. J. Biol. Chem. 272, 233–239.

17.	Schoeni, J. L. and Wong, A. C. (1999) Heterogeneity observed in the components of hemolysin BL, an enterotoxin produced by Bacillus cereus. Int. J. Food Microbiol. 53, 159–167.

18.	Beecher, D. J., Pulido, J. S., Barney, N. P., and Wong, A. C. (1995) Extracellular virulence factors in Bacillus cereus endophthalmitis: methods and implication of involvement of hemolysin BL. Infect. Immunol. 63, 632–639.

19.	Lund, T. and Granum, P. E. (1996) Characterisation of a non-haemolytic enterotoxin complex from Bacillus cereus isolated after a foodborne outbreak. FEMS Microbiol. Lett. 141, 151–156.

20.	Gaviria Rivera, A. M., Granum, P. E., and Priest, F. G. (2000) Common occurrence of enterotoxin genes and enterotoxicity in Bacillus thuringiensis. FEMS Microbiol. Lett. 190, 151–155.

21.	Guinebretiere, M. H., Broussolle, V., and Nguyen-The, C. (2002) Enterotoxigenic profiles of food-poisoning and food-borne Bacillus cereus strains. J. Clin. Microbiol. 40, 3053–3056.

22.	Lund, T., De Buyser, M. L., and Granum, P. E. (2000) A new cytotoxin from Bacillus cereus that may cause necrotic enteritis. Mol. Microbiol. 38, 254–261.

23.	Agata, N., Ohta, M., Arakawa, Y., and Mori, M. (1995) The bceT gene of Bacillus cereus encodes an enterotoxic protein. Microbiology 141, 983–988.

24.	Asano, S. I., Nukumizu, Y., Bando, H., Iizuka, T., and Yamamoto, T. (1997) Cloning of novel enterotoxin genes from Bacillus cereus and Bacillus thuringiensis. Appl. Environ. Microbiol. 63, 1054–1057.

25.	Choma, C. and Granum, P. E. (2002) The enterotoxin T (BcET) from Bacillus cereus can probably not contribute to food poisoning. FEMS Microbiol. Lett. 217, 115–119.

26.	Hansen, B. M., Hoiby, P. E., Jensen, G. B., and Hendriksen, N. B. (2003) The Bacillus cereus bceT enterotoxin sequence reappraised. FEMS Microbiol. Lett. 223, 21–24.

27.	Margot, P., Wahlen, M., Gholamhoseinian, A., Piggot, P., Karamata, D., and Gholamhuseinian, A. (1998) The lytE gene of Bacillus subtilis 168 encodes a cell wall hydrolase. J. Bacteriol. 180, 749–752.

28.	Agata, N., Mori, M., Ohta, M., Suwan, S., Ohtani, I., and Isobe. M. (1994) A novel dodecadepsipeptide, cereulide, isolated from Bacillus cereus causes vacuole formation in HEp-2 cells. FEMS Microbiol. Lett. 121, 31–34.

29.	Shinagawa, K., Konuma, H., Sekita, H., and Sugii, S. (1995) Emesis of rhesus monkeys induced by intragastric administration with the HEp-2 vacuolation factor (cereulide) produced by Bacillus cereus. FEMS Microbiol. Lett. 130, 87–90.

30.	Agata, N., Ohta, M., Mori, M., and Isobe, M. (1995) A novel dodecadepsipeptide, cereulide, is an emetic toxin of Bacillus cereus. FEMS Microbiol. Lett. 129, 17–20.

31.	Gilmore, M. S., Cruz-Rodz, A. L., Leimeister-Wachter, M., Kreft, J., and Goebel, W. (1989) A Bacillus cereus cytolytic determinant, cereolysin AB, which comprises the phospholipase C and sphingomyelinase genes: nucleotide sequence and genetic linkage. J. Bacteriol. 171, 744–753.

32.	Volwerk, J. J., Wetherwax, P. B., Evans, L. M., Kuppe, A., and Griffith, O. H. (1989) Phosphatidylinositol-specific phospholipase C from Bacillus cereus: improved purification, amino acid composition, and amino-terminal sequence. J. Cell. Biochem. 39, 315–325.

33.	Baida, G. E. and Kuzmin, N. P. (1995) Cloning and primary structure of a new hemolysin gene from Bacillus cereus. Biochem. Biophys. Acta 1264, 151–154.

34.	Honda, T., Shiba, A., Seo, S., Yamamoto, J., Matsuyama, J., and Miwatani, T. (1991) Identity of hemolysins produced by Bacillus thuringiensis and Bacillus Cereus. FEMS Microbiol. Lett. 63, 205–209.

35.	Sinev, M. A., Budarina, Z., Gavrilenko, I. V., Tomashevskii, A. I., and Kuzmin, N. P. (1993) Evidence of the existence of hemolysin II from Bacillus cereus: cloning the genetic determinant of hemolysin II. Mol. Biol. (Mosk) 27, 1218–1229.

36.	Pospiech, A. and Neumann, B. (1995) A versatile quick-prep of genomic DNA from Gram-positive bacteria. Trends Genet. 11, 217–218.

37.	Beecher, D. J. and Wong, A. C. (1994) Identification and analysis of the antigens detected by two commercial Bacillus cereus diarrhoeal enterotoxin immunoassay kits. Appl. Environ. Microbiol. 60, 4614–4616.

38.	Granum, P. E., Brynestad, S., and Kramer, J. M. (1993) Analysis of enterotoxin production by Bacillus cereus from dairy products, food poisoning incidents and non-gastrointestinal infections. Int. J. Food Microbiol. 17, 269–279.

39.	Lund, T. and Granum, P. E. (1997) Comparison of biological effect of the two different enterotoxin complexes isolated from three different strains of Bacillus cereus. Microbiology 143, 3329–3336.

40.	Jääskeläinen, E. L., Teplova, V., Andersson, M. A., et al. (2003) In vitro assay for human toxicity of cereulide, the emetic mitochondrial toxin produced by food poisoning Bacillus cereus. Toxicology in Vitro 17, 737–744.

41.	Häggblom, M. M., Apetroaie, C., Andersson, M. A., and Salkinoja-Salonen, M. S. (2002) Quantitative analysis of cereulide, the emetic toxin of Bacillus cereus, produced under various conditions. Appl. Environ. Microbiol. 68, 2479–2483.

Comments (Loading...)

Post comment/View all comments