Inter-Research > DAO > v155 > p141-146  
Diseases of Aquatic Organisms

via Mailchimp

DAO 155:141-146 (2023)  -  DOI:

Comparison of methods for determining Batrachochytrium dendrobatidis zoospore viability

Adeline Chew1, Matt West1, Laura A. Brannelly2,*

1School of Biosciences, Faculty of Science, The University of Melbourne, Parkville, Victoria 3010, Australia
2Melbourne Veterinary School, Faculty of Sciences, The University of Melbourne, Werribee, Victoria 3030, Australia
*Corresponding author:

ABSTRACT: The emerging fungal pathogen Batrachochytrium dendrobatidis (Bd) threatens hundreds of amphibian species globally. During laboratory-based experiments it is often essential to quantify live Bd cells, but a comparison of the effectiveness of methods for counting and assessing the viability of the infectious zoospore life stage has not been done. A direct comparison of staining methods that assess viability will ensure that the most accurate and efficient method is used. Here, we compared the use of 2 relatively cheap common stains, trypan blue and methylene blue, and assessed their accuracy and precision for estimating the viability of Bd zoospores during both manual counting and colorimetric assays. We stained known proportions of killed Bd zoospores (0, 0.25, 0.50, 0.75, and 1.00) with each stain and estimated the proportion of stained (dead) and unstained (viable) cells in each sample using both manual counting and colorimetric assays. Trypan blue was found to be a much more effective stain than methylene blue for both microscopy and colorimetric assays. Additionally, counting zoospores via microscopy was both a more accurate and precise technique. We recommend using manual counts via microscopy using the trypan blue stain for assessing Bd zoospore viability.

KEY WORDS: Batrachochytrium dendrobatidis · Chytridiomycosis · Cell viability · Trypan blue · Methylene blue · Spectrophotometer · Counts

Full text in pdf format
Cite this article as: Chew A, West M, Brannelly LA (2023) Comparison of methods for determining Batrachochytrium dendrobatidis zoospore viability. Dis Aquat Org 155:141-146.

Export citation
Share:    Facebook - - linkedIn

 Previous article Next article