Efficacy of SYBR 14/propidium iodide viability stain for the amphibian chytrid fungus Batrachochytrium dendrobatidis.
The amphibian chytrid fungus Batrachochytrium dendrobatidis is a recently described pathogen that has been implicated as a causal agent in the global decline in amphibians. Research into its biology and epidemiology has frequently involved in vitro experimentation. However, this research is currently limited by the inability to differentiate between viable and inviable zoospores. Stains are frequently used to determine cell viability, and this study tested a 2-colour fluorescence assay for the detection and quantification of viable B. dendrobatidis zoospores. The results show that the nucleic acid stains SYBR 14 and propidium iodide are effective in distinguishing live from dead zoospores, and a protocol has been optimized for their use. This viability assay provides an efficient and reliable tool that will have applications in B. dendrobatidis challenge and amphibian exposure experiments.
School of Environmental and Life Sciences, University of Newcastle, Callaghan Drive, Callaghan, New South Wales 2308, Australia. michelle.stockwell@newcastle.edu.au
M P
MP
Stockwell
J
J
Clulow
M J
MJ
Mahony
D000663:Amphibians / Q000382:microbiology; D000818:Animals; D008411:Chytridiomycota / Q000302:isolation & purification*; D007202:Indicators and Reagents; D009930:Organic Chemicals; D011419:Propidium; D013194:Staining and Labeling
D007202:Indicators and Reagents; D009930:Organic Chemicals; C093207:SYBR-14; D011419:Propidium