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

Fisher, Matthew C., Ghosh, Pria, Shelton, Jennifer M.G., Bates, Kieran, Brookes, Lola, Wierzbicki, Claudia, Rosa, Gonçalo M., Farrer, Rhys A., Aanensen, David M., Alvarado-Rybak, Mario, Bataille, Arnaud, Berger, Lee, Böll, Susanne, Bosch, Jaime, Clare, Frances C., Courtois, Elodie A., Crottini, Angelica, Cunningham, Andrew A., Doherty-Bone, Thomas M., Gebresenbet, Fikirte, Gower, David J., Höglund, Jacob, James, Timothy Y., Jenkinson, Thomas S., Kosch, Tiffany A., Lambertini, Carolina, Laurila, Anssi, Lin, Chun-Fu, Loyau, Adeline, Martel, An, Meurling, Sara, Miaud, Claude, Minting, Pete, Ndriantsoa, Serge, O'Hanlon, Simon J., Pasmans, Frank, Rakotonanahary, Tsanta, Rabemananjara, Falitiana C.E., Ribeiro, Luisa P., Schmeller, Dirk S., Schmidt, Benedikt R., Skerratt, Lee, Smith, Freya, Soto-Azat, Claudio, Tessa, Giulia, Toledo, Luís Felipe, Valenzuela-Sánchez, Andrés, Verster, Ruhan, Vörös, Judit, Waldman, Bruce, Webb, Rebecca J., Weldon, Che, Wombwell, Emma, Zamudio, Kelly R., Longcore, Joyce E., and Garner, Trenton W.J. (2018) Development and worldwide use of non-lethal, and minimal population-level impact, protocols for the isolation of amphibian chytrid fungi. Scientific Reports, 8. 7772.

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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.

Item ID: 53874
Item Type: Article (Research - C1)
ISSN: 2045-2322
Copyright Information: Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
Funders: National Environmental Research Council (NERC), Leverhulme Trust (LT), UK, Morris Animal Foundation (MAF), Hunagrian Academy of Sciences (HAS), Conservation Leadership Programme Future Conservationist Award, Fondecyt, Mohamed bin Zayed Species Conservation Fund (MbZ), Fundação para a Ciência e a Tecnologia, Portugal (FCT), São Paulo Research Foundation (FAPESP), National Counsel of Technological and Scientific Development (CNPq), National Science Foundation (NSF), USA, Royal Geographical Society, Royal Zoological Society of Scotland, National Research Foundation of Korea (NRF)
Projects and Grants: NERC grant NE/K014455/1, NERC grant NE/E006701/1, NERC grant ANR-08-BDVA-002-03, LT RPG-2014-273, HAS Bolyai János Research Grant BO/00597/14, Fondecyt 1181758, MbZ project 152510704, FCT scholarship SFRH/BD/69194/2010, FAPESP #2016/25358-3, CNPq #300896/2016-6, NSF OISE-1159513, NRF 2015R1D1A1A01057282
Date Deposited: 06 Jun 2018 07:36
FoR Codes: 07 AGRICULTURAL AND VETERINARY SCIENCES > 0707 Veterinary Sciences > 070704 Veterinary Epidemiology @ 100%
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