TY - JOUR T1 - Development and worldwide use of a non-lethal and minimal population-level impact protocols for the isolation of chytrids from amphibians JF - bioRxiv DO - 10.1101/246538 SP - 246538 AU - Matthew C. Fisher AU - Pria Ghosh AU - Jennifer M. G. Shelton AU - Kieran Bates AU - Lola Brookes AU - Claudia Wierzbicki AU - Gonçalo M. Rosa AU - Rhys A. Farrer AU - David M. Aanensen AU - Mario Alvarado-Rybak AU - Arnaud Bataille AU - Lee Berger AU - Susanne Böll AU - Jaime Bosch AU - France C. Clare AU - Elodie Courtois AU - Angelica Crottini AU - Andrew A. Cunningham AU - Thomas M. Doherty-Bone AU - Fikirte Gebresenbet AU - David J. Gower AU - Jacob Höglund AU - Timothy Y. James AU - Thomas S. Jenkinson AU - Tiffany A. Kosch AU - Carolina Lambertini AU - Anssi Laurila AU - Chun-Fu Lin AU - Adeline Loyau AU - An Martel AU - Sara Meurling AU - Claude Miaud AU - Pete Minting AU - Serge Ndriantsoa AU - Frank Pasmans AU - Tsanta Rakotonanahary AU - Falitiana C. E. Rabemananjara AU - Luisa P. Ribeiro AU - Dirk S. Schmeller AU - Benedikt R. Schmidt AU - Lee Skerratt AU - Freya Smith AU - Claudio Soto-Azat AU - Giulia Tessa AU - Luís Felipe Toledo AU - Andrés Valenzuela-Sánchez AU - Ruhan Verster AU - Judit Vörös AU - Bruce Waldman AU - Rebecca J. Webb AU - Che Weldon AU - Emma Wombwell AU - Kelly R. Zamudio AU - Joyce E. Longcore AU - Trenton W.J. Garner Y1 - 2018/01/01 UR - http://biorxiv.org/content/early/2018/01/14/246538.abstract N2 - Parasitic chytrid fungi have emerged as a significant threat to amphibian species worldwide, necessitating the development of techniques to isolate these pathogens into sterile 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 interested researchers worldwide 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 widely applied across at least 5 continents, 23 countries and in 62 amphibian species, and have been successfully used to isolate chytrids in remote field locations. 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 important group of emerging fungal pathogens. ER -