Development and worldwide use of non-lethal, and minimal population-level impact, protocols for the isolation of amphibian chytrid fungi

Matthew C Fisher (Corresponding Author), Pria Ghosh, Jennifer M G Shelton, Kieran Bates, Lola Brookes, Claudia Wierzbicki, Gonçalo M Rosa, Rhys A Farrer, David M Aanensen, Mario Alvarado-Rybak, Arnaud Bataille, Lee Berger, Susanne Böll, Jaime Bosch, Frances C Clare, Elodie A Courtois, Angelica Crottini, Andrew A Cunningham, Thomas M Doherty-Bone, Fikirte GebresenbetDavid J Gower, Jacob Höglund, Timothy Y James, Thomas S Jenkinson, Tiffany A Kosch, Carolina Lambertini, Anssi Laurila, Chun-Fu Lin, Adeline Loyau, An Martel, Sara Meurling, Claude Miaud, Pete Minting, Serge Ndriantsoa, Simon J O'Hanlon, Frank Pasmans, Tsanta Rakotonanahary, Falitiana C E Rabemananjara, Luisa P Ribeiro, Dirk S Schmeller, Benedikt R Schmidt, Lee Skerratt, Freya Smith, Claudio Soto-Azat, Giulia Tessa, Luís Felipe Toledo, Andrés Valenzuela-Sánchez, Ruhan Verster, Judit Vörös, Bruce Waldman, Rebecca J Webb, Che Weldon, Emma Wombwell, Kelly R Zamudio, Joyce E Longcore, Trenton W J Garner

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Abstract

Parasitic chytrid fungi have emerged as a significant threat to amphibian species worldwide, necessitating the development of techniques to isolate these pathogens into culture for research purposes. However, early methods of isolating chytrids from their hosts relied on killing amphibians. We modified a pre-existing protocol for isolating chytrids from infected animals to use toe clips and biopsies from toe webbing rather than euthanizing hosts, and distributed the protocol to researchers as part of the BiodivERsA project RACE; here called the RML protocol. In tandem, we developed a lethal procedure for isolating chytrids from tadpole mouthparts. Reviewing a database of use a decade after their inception, we find that these methods have been applied across 5 continents, 23 countries and in 62 amphibian species. Isolation of chytrids by the non-lethal RML protocol occured in 18% of attempts with 207 fungal isolates and three species of chytrid being recovered. Isolation of chytrids from tadpoles occured in 43% of attempts with 334 fungal isolates of one species (Batrachochytrium dendrobatidis) being recovered. Together, these methods have resulted in a significant reduction and refinement of our use of threatened amphibian species and have improved our ability to work with this group of emerging pathogens.

Original languageEnglish
Article number7772
JournalScientific Reports
Volume8
DOIs
Publication statusPublished - 17 May 2018

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Keywords

  • biological techniques
  • ecological epidemiology
  • microbiology

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