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Journal of Wildlife Diseases, 44(2), 2008, pp. 209-225
© Wildlife Disease Association  2008
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USING AMPLIFIED FRAGMENT LENGTH POLYMORPHISM ANALYSIS TO DIFFERENTIATE ISOLATES OF PASTEURELLA MULTOCIDA SEROTYPE 1

 David S. Blehert1,3, Keynttisha L. Jefferson1, Dennis M. Heisey1, Michael D. Samuel2, Brenda M. Berlowski1 and Daniel J. Shadduck1

1 US Geological Survey, National Wildlife Health Center, 6006 Schroeder Road, Madison, Wisconsin 53711, USA
2 US Geological Survey, Wisconsin Cooperative Wildlife Research Unit, Department of Wildlife Ecology, 204 Russell Labs, 1630 Linden Drive, Madison, Wisconsin 53706, USA
3 Corresponding author (email: dblehert{at}usgs.gov)

ABSTRACT:   Avian cholera, an infectious disease caused by the bacterium Pasteurella multocida, kills thousands of North American wild waterfowl annually. Pasteurella multocida serotype 1 isolates cultured during a laboratory challenge study of Mallards (Anas platyrhynchos) and collected from wild birds and environmental samples during avian cholera outbreaks were characterized using amplified fragment length polymorphism (AFLP) analysis, a whole-genome DNA fingerprinting technique. Comparison of the AFLP profiles of 53 isolates from the laboratory challenge demonstrated that P. multocida underwent genetic changes during a 3-mo period. Analysis of 120 P. multocida serotype 1 isolates collected from wild birds and environmental samples revealed that isolates were distinguishable from one another based on regional and temporal genetic characteristics. Thus, AFLP analysis had the ability to distinguish P. multocida isolates of the same serotype by detecting spatiotemporal genetic changes and provides a tool to advance the study of avian cholera epidemiology. Further application of AFLP technology to the examination of wild bird avian cholera outbreaks may facilitate more effective management of this disease by providing the potential to investigate correlations between virulence and P. multocida genotypes, to identify affiliations between bird species and bacterial genotypes, and to elucidate the role of specific bird species in disease transmission.
  Key words:  Amplified fragment length polymorphism, avian cholera, DNA fingerprinting, Pasteurella multocida, waterfowl.






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