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Diseases of Aquatic Organisms

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DAO 96:169-174 (2011)  -  DOI:

Fluorescent probes as a tool for labelling and tracking the amphibian chytrid fungus Batrachochytrium dendrobatidis

Sarah M. Herbert1,2,*, Tommy L. F. Leung1,3, Phillip J. Bishop1

1Department of Zoology, University of Otago, Dunedin 9016, New Zealand
2Present address: EcoGecko Consultants Ltd, Wellington 6012, New Zealand
3Present address: Zoology, School of Environmental and Rural Science, University of New England, Armidale, New South Wales 2351, Australia

ABSTRACT: The dissemination of the virulent pathogen Batrachochytrium dendrobatidis (Bd) has contributed to the decline and extinction of many amphibian species worldwide. Several different strains have been identified, some of which are sympatric. Interactions between co-infecting strains of a pathogen can have significant influences on disease epidemiology and evolution; therefore the dynamics of multi-strain infections is an important area of research. We stained Bd cells with 2 fluorescent BODIPY® fatty acid probes to determine whether these can potentially be used to distinguish and track Bd cell lines in multi-strain experiments. Bd cells in broth culture were stained with 5 concentrations of green-fluorescent BODIPY FL and red-fluorescent BODIPY 558/568 and visualised under an epifluorescent microscope for up to 16 d post-dye. Dyed strains were also assessed for growth inhibition. The most effective concentration for both dyes was 10 µM. This concentration of dye produced strong fluorescence for 12 to 16 d in Bd cultures held at 23°C (3 to 4 generations), and did not inhibit Bd growth. Cells dyed with BODIPY FL and BODIPY 558/568 can be distinguished from each other on the basis of their fluorescence characteristics. Therefore, it is likely that this technique will be useful for research into multi-strain dynamics of Bd infections.

KEY WORDS: Amphibian chytrid fungus · Batrachochytrium dendrobatidis · Epifluorescence microscopy · Co-infection · BODIPY FL · BODIPY 558/568

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Cite this article as: Herbert SM, Leung TLF, Bishop PJ (2011) Fluorescent probes as a tool for labelling and tracking the amphibian chytrid fungus Batrachochytrium dendrobatidis. Dis Aquat Org 96:169-174.

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