DAO 48:213-220 (2002)  -  doi:10.3354/dao048213

Production of polyclonal antibodies to Batrachochytrium dendrobatidis and their use in an immunoperoxidase test for chytridiomycosis in amphibians

Lee Berger1, Alex D. Hyatt1,*, Veronica Olsen1, Sandra G. Hengstberger1, Donna Boyle1, Gerry Marantelli2, Kaye Humphreys3, Joyce E. Longcore4

1Australian Animal Health Laboratory, CSIRO Livestock Industries, Private Bag 24, Geelong, Victoria 3220, Australia
2Amphibian Research Centre, PO Box 424, Brunswick, Victoria 3056, Australia
3Veterinary and Quarantine Centre, Taronga Zoo, PO Box 20, Mosman, New South Wales 2088, Australia
4Department of Biological Sciences, University of Maine, Orono, Maine 04469-5722, USA

ABSTRACT: Polyclonal antibodies were produced for diagnosing chytridiomycosis in amphibians. Two sheep and 4 rabbits were inoculated with homogenized whole culture of Batrachochytrium dendrobatidis in Freund¹s complete adjuvant or triple adjuvant. Antisera from all animals reacted strongly with all stages of B. dendrobatidis and stained the walls, cytoplasm, rhizoids and zoospores in an indirect immunoperoxidase test. Significant cross-reactivity occurred only with some fungi in the Chytridiomycota, and there are no members of this phylum besides B. dendrobatidis that infect frogs. The immunoperoxidase stain is a useful screening test when combined with recognition of the morphology and infection site of B. dendrobatidis.


KEY WORDS: Batrachochytrium dendrobatidis · Chytridiomycosis · Fungus · Amphibians · Immunoperoxidase · Polyclonal antibodies · Diagnosis


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