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

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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.
Keywords
BATRACHOCHYTRIUM-DENDROBATIDIS, ORAL CHYTRIDIOMYCOSIS, SP-NOV, PATHOGEN, SALAMANDRIVORANS, INFECTION, DECLINES, TRADE, TIME, EXTINCTIONS

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MLA
Fisher, Matthew C et al. “Development and Worldwide Use of Non-lethal, and Minimal Population-level Impact, Protocols for the Isolation of Amphibian Chytrid Fungi.” SCIENTIFIC REPORTS 8 (2018): n. pag. Print.
APA
Fisher, M. C., Ghosh, P., Shelton, J. M., Bates, K., Brookes, L., Wierzbicki, C., Rosa, G. M., et al. (2018). Development and worldwide use of non-lethal, and minimal population-level impact, protocols for the isolation of amphibian chytrid fungi. SCIENTIFIC REPORTS, 8.
Chicago author-date
Fisher, Matthew C, Pria Ghosh, Jennifer MG Shelton, Kieran Bates, Lola Brookes, Claudia Wierzbicki, Goncalo M Rosa, et al. 2018. “Development and Worldwide Use of Non-lethal, and Minimal Population-level Impact, Protocols for the Isolation of Amphibian Chytrid Fungi.” Scientific Reports 8.
Chicago author-date (all authors)
Fisher, Matthew C, Pria Ghosh, Jennifer MG Shelton, Kieran Bates, Lola Brookes, Claudia Wierzbicki, Goncalo M Rosa, Rhys A Farrer, David M Aanensen, Mario Alvarado-Rybak, Arnaud Bataille, Lee Berger, Susanne Boell, Jaime Bosch, Frances C Clare, Elodie A Courtois, Angelica Crottini, Andrew A Cunningham, Thomas M Doherty-Bone, Fikirte Gebresenbet, David J Gower, Jacob Hoglund, Timothy Y James, Thomas S Jenkinson, Tiffany A Kosch, Carolina Lambertini, Anssi Laurila, Chun-Fu Lin, Adeline Loyau, An Martel, Sar Meurling, Claude Miaud, Pete Minting, Serge Ndriantsoa, Simon J O’Hanlon, Frank Pasmans, Tsanta Rakotonanahary, Falitiana CE Rabemananjara, Luisa P Ribeiro, Dirk S Schmeller, Benedikt R Schmidt, Lee Skerratt, Freya Smith, Claudio Soto-Azat, Giulia Tessa, Luis Felipe Toledo, Andres Valenzuela-Sanchez, Ruhan Verster, Judit Voeroes, Bruce Waldman, Rebecca J Webb, Che Weldon, Emma Wombwell, Kelly R Zamudio, Joyce E Longcore, and Trenton WJ Garner. 2018. “Development and Worldwide Use of Non-lethal, and Minimal Population-level Impact, Protocols for the Isolation of Amphibian Chytrid Fungi.” Scientific Reports 8.
Vancouver
1.
Fisher MC, Ghosh P, Shelton JM, Bates K, Brookes L, Wierzbicki C, et al. Development and worldwide use of non-lethal, and minimal population-level impact, protocols for the isolation of amphibian chytrid fungi. SCIENTIFIC REPORTS. 2018;8.
IEEE
[1]
M. C. Fisher et al., “Development and worldwide use of non-lethal, and minimal population-level impact, protocols for the isolation of amphibian chytrid fungi,” SCIENTIFIC REPORTS, vol. 8, 2018.
@article{8610490,
  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.},
  articleno    = {7772},
  author       = {Fisher, Matthew C and Ghosh, Pria and Shelton, Jennifer MG and Bates, Kieran and Brookes, Lola and Wierzbicki, Claudia and Rosa, Goncalo M and Farrer, Rhys A and Aanensen, David M and Alvarado-Rybak, Mario and Bataille, Arnaud and Berger, Lee and Boell, Susanne and Bosch, Jaime and Clare, Frances C and Courtois, Elodie A and Crottini, Angelica and Cunningham, Andrew A and Doherty-Bone, Thomas M and Gebresenbet, Fikirte and Gower, David J and Hoglund, Jacob and James, Timothy Y and Jenkinson, Thomas S and Kosch, Tiffany A and Lambertini, Carolina and Laurila, Anssi and Lin, Chun-Fu and Loyau, Adeline and Martel, An and Meurling, Sar and Miaud, Claude and Minting, Pete and Ndriantsoa, Serge and O'Hanlon, Simon J and Pasmans, Frank and Rakotonanahary, Tsanta and Rabemananjara, Falitiana CE and Ribeiro, Luisa P and Schmeller, Dirk S and Schmidt, Benedikt R and Skerratt, Lee and Smith, Freya and Soto-Azat, Claudio and Tessa, Giulia and Toledo, Luis Felipe and Valenzuela-Sanchez, Andres and Verster, Ruhan and Voeroes, Judit and Waldman, Bruce and Webb, Rebecca J and Weldon, Che and Wombwell, Emma and Zamudio, Kelly R and Longcore, Joyce E and Garner, Trenton WJ},
  issn         = {2045-2322},
  journal      = {SCIENTIFIC REPORTS},
  keywords     = {BATRACHOCHYTRIUM-DENDROBATIDIS,ORAL CHYTRIDIOMYCOSIS,SP-NOV,PATHOGEN,SALAMANDRIVORANS,INFECTION,DECLINES,TRADE,TIME,EXTINCTIONS},
  language     = {eng},
  pages        = {8},
  title        = {Development and worldwide use of non-lethal, and minimal population-level impact, protocols for the isolation of amphibian chytrid fungi},
  url          = {http://dx.doi.org/10.1038/s41598-018-24472-2},
  volume       = {8},
  year         = {2018},
}

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