Title: Detection of Clostridium botulinum type C in an aquatic area a year after an outbreak of botulism in waterfowl using conventional and molecular methods

Authors: Matjaz Ocepek, Aleksandra Hari, Branko Krt, Mateja Pate, Irena Zdovc

Addresses: Veterinary Faculty, Institute of Microbiology and Parasitology, University of Ljubljana, Gerbiceva 60, SI 1000 Ljubljana, Slovenia. ' Veterinary Faculty, Institute of Microbiology and Parasitology, University of Ljubljana, Gerbiceva 60, SI 1000 Ljubljana, Slovenia. ' Veterinary Faculty, Institute of Microbiology and Parasitology, University of Ljubljana, Gerbiceva 60, SI 1000 Ljubljana, Slovenia. ' Veterinary Faculty, Institute of Microbiology and Parasitology, University of Ljubljana, Gerbiceva 60, SI 1000 Ljubljana, Slovenia ' Veterinary Faculty, Institute of Microbiology and Parasitology, University of Ljubljana, Gerbiceva 60, SI 1000 Ljubljana, Slovenia

Abstract: A year after an outbreak of botulism in waterfowl, the presence of the causative agent of the disease, Clostridium botulinum type C, was investigated, using bioassay and polymerase chain reaction (PCR). Eighty two mud samples collected at different locations in the Skocjanski zatok nature reserve were investigated. Mouse bioassay revealed that 38 (46.4%) samples contained C. botulinum type C neurotoxin. PCR targeted at the C. botulinum type C neurotoxin gene revealed six (7.3%) positive samples. The number of C. botulinum spores in 1 g of mud was also estimated. We did not manage to isolate C. botulinum from mud samples, but we did isolate one type C neurotoxin producing strain from an animal that died during the outbreak. The toxin type was established in the mouse toxin neutralisation test and the strain was identified with PCR.

Keywords: microbiology; veterinary; Clostridium botulinum; isolation; identification; botulism; diagnosis; mouse bioassay; polymerase chain reaction; waterfowl diseases; Slovenia.

DOI: 10.1504/IJEP.2007.015664

International Journal of Environment and Pollution, 2007 Vol.31 No.1/2, pp.65 - 74

Published online: 07 Nov 2007 *

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