Abstract
A PCR was developed and applied for the detection of Clostridium botulinum type C in 18 avian and environmental samples collected during an outbreak of avian botulism, and the results were compared with those obtained by conventional methodologies based on the mouse bioassay. PCR and mouse bioassay results compared well (100%) after the enrichment of samples, but PCR results directly indicated the presence of this microorganism in six samples, while only one of these contained the type C botulinal neurotoxin before enrichment. The PCR assay was sensitive (limit of detection between 15 and 15 x 10(3) spores per PCR), specific (no amplification products were obtained with other clostridia), and rapid, since sonicated and heated samples provided enough template for amplification without any DNA purification. Eleven isolates of C. botulinum type C were recovered from mallards (Anas platyrhynchos), grey herons (Ardea cinerea), and mud during investigation of this outbreak.
Full Text
The Full Text of this article is available as a PDF (190.3 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Aureli P., Fenicia L., Pasolini B., Gianfranceschi M., McCroskey L. M., Hatheway C. L. Two cases of type E infant botulism caused by neurotoxigenic Clostridium butyricum in Italy. J Infect Dis. 1986 Aug;154(2):207–211. doi: 10.1093/infdis/154.2.207. [DOI] [PubMed] [Google Scholar]
- DasGupta B. R. Structure and biological activity of botulinum neurotoxin. J Physiol (Paris) 1990;84(3):220–228. [PubMed] [Google Scholar]
- Duddle M. Emotional sequelae of sexual assault. J R Soc Med. 1991 Jan;84(1):26–28. doi: 10.1177/014107689108400110. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Eklund M. W., Poysky F. T., Reed S. M., Smith C. A. Bacteriophage and the toxigenicity of Clostridium botulinum type C. Science. 1971 Apr 30;172(3982):480–482. doi: 10.1126/science.172.3982.480. [DOI] [PubMed] [Google Scholar]
- Fach P., Hauser D., Guillou J. P., Popoff M. R. Polymerase chain reaction for the rapid identification of Clostridium botulinum type A strains and detection in food samples. J Appl Bacteriol. 1993 Sep;75(3):234–239. doi: 10.1111/j.1365-2672.1993.tb02771.x. [DOI] [PubMed] [Google Scholar]
- Franciosa G., Ferreira J. L., Hatheway C. L. Detection of type A, B, and E botulism neurotoxin genes in Clostridium botulinum and other Clostridium species by PCR: evidence of unexpressed type B toxin genes in type A toxigenic organisms. J Clin Microbiol. 1994 Aug;32(8):1911–1917. doi: 10.1128/jcm.32.8.1911-1917.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Habermann E., Dreyer F. Clostridial neurotoxins: handling and action at the cellular and molecular level. Curr Top Microbiol Immunol. 1986;129:93–179. doi: 10.1007/978-3-642-71399-6_2. [DOI] [PubMed] [Google Scholar]
- Hauser D., Eklund M. W., Kurazono H., Binz T., Niemann H., Gill D. M., Boquet P., Popoff M. R. Nucleotide sequence of Clostridium botulinum C1 neurotoxin. Nucleic Acids Res. 1990 Aug 25;18(16):4924–4924. doi: 10.1093/nar/18.16.4924. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Montecucco C., Schiavo G. Mechanism of action of tetanus and botulinum neurotoxins. Mol Microbiol. 1994 Jul;13(1):1–8. doi: 10.1111/j.1365-2958.1994.tb00396.x. [DOI] [PubMed] [Google Scholar]
- Oguma K., Yamaguchi T., Sudou K., Yokosawa N., Fujikawa Y. Biochemical classification of Clostridium botulinum type C and D strains and their nontoxigenic derivatives. Appl Environ Microbiol. 1986 Feb;51(2):256–260. doi: 10.1128/aem.51.2.256-260.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schiavo G., Shone C. C., Bennett M. K., Scheller R. H., Montecucco C. Botulinum neurotoxin type C cleaves a single Lys-Ala bond within the carboxyl-terminal region of syntaxins. J Biol Chem. 1995 May 5;270(18):10566–10570. doi: 10.1074/jbc.270.18.10566. [DOI] [PubMed] [Google Scholar]
- Szabo E. A., Pemberton J. M., Gibson A. M., Eyles M. J., Desmarchelier P. M. Polymerase chain reaction for detection of Clostridium botulinum types A, B and E in food, soil and infant faeces. J Appl Bacteriol. 1994 Jun;76(6):539–545. doi: 10.1111/j.1365-2672.1994.tb01650.x. [DOI] [PubMed] [Google Scholar]
- Szabo E. A., Pemberton J. M., Gibson A. M., Thomas R. J., Pascoe R. R., Desmarchelier P. M. Application of PCR to a clinical and environmental investigation of a case of equine botulism. J Clin Microbiol. 1994 Aug;32(8):1986–1991. doi: 10.1128/jcm.32.8.1986-1991.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]