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

Author(s):Show less -

Fisher, Matthew C. ^[1] ; Ghosh, Pria ^{[1, 2]} ; Shelton, Jennifer M. G. ^[1] ; Bates, Kieran ^[1] ; Brookes, Lola ^[3] ; Wierzbicki, Claudia ^[1] ; Rosa, Goncalo M. ^{[3, 4]} ; Farrer, Rhys A. ^[1] ; Aanensen, David M. ^{[1, 5]} ; Alvarado-Rybak, Mario ^[6] ; Bataille, Arnaud ^{[7, 8, 9]} ; Berger, Lee ^[10] ; Boell, Susanne ^[11] ; Bosch, Jaime ^[12] ; Clare, Frances C. ^[1] ; Courtois, Elodie A. ^[13] ; Crottini, Angelica ^[14] ; Cunningham, Andrew A. ^[3] ; Doherty-Bone, Thomas M. ^[15] ; Gebresenbet, Fikirte ^[16] ; Gower, David J. ^[17] ; Hoglund, Jacob ^[18] ; James, Timothy Y. ^[19] ; Jenkinson, Thomas S. ^[19] ; Kosch, Tiffany A. ^{[7, 10]} ; Lambertini, Carolina ^[20] ; Laurila, Anssi ^[18] ; Lin, Chun-Fu ^[21] ; Loyau, Adeline ^{[22, 23]} ; Martel, An ^[24] ; Meurling, Sara ^[18] ; Miaud, Claude ^[25] ; Minting, Pete ^[26] ; Ndriantsoa, Serge ^[27] ; O'Hanlon, Simon J. ^{[3, 1]} ; Pasmans, Frank ^[24] ; Rakotonanahary, Tsanta ^[27] ; Rabemananjara, Falitiana C. E. ^{[27, 28]} ; Ribeiro, Luisa P. ^[20] ; Schmeller, Dirk S. ^{[22, 23]} ; Schmidt, Benedikt R. ^{[29, 30]} ; Skerratt, Lee ^[10] ; Smith, Freya ^[31] ; Soto-Azat, Claudio ^[6] ; Tessa, Giulia ^[32] ; Toledo, Luis Felipe ^[20] ; Valenzuela-Sanchez, Andres ^{[6, 33]} ; Verster, Ruhan ^[2] ; Voeroes, Judit ^[34] ; Waldman, Bruce ^[7] ; Webb, Rebecca J. ^[10] ; Weldon, Che ^[2] ; Wombwell, Emma ^[3] ; Zamudio, Kelly R. ^[35] ; Longcore, Joyce E. ^[36] ; Garner, Trenton W. J. ^{[3, 2, 32]}

Affiliation:Show less -

^[1] Imperial Coll London, Sch Publ Hlth, Fac Med, Dept Infect Dis Epidemiol, St Marys Campus, London W2 1PG - England

^[2] North West Univ, Unit Environm Sci & Management, Private Bag x6001, ZA-2520 Potchefstroom - South Africa

^[3] Inst Zool, Regents Pk, London NW1 4RY - England

^[4] Univ Lisbon, Fac Ciencias, Ctr Ecol Evolut & Environm Changes CE3C, Lisbon - Portugal

^[5] Ctr Genom Pathogen Surveillance, Wellcome Genome Campus, Hinxton, Cambs - England

^[6] Univ Andres Bello, Fac Ecol & Recursos Nat, Ctr Invest Sustentabilidad, Republ 440, Santiago - Chile

^[7] Seoul Natl Univ, Sch Biol Sci, Lab Behav & Populat Ecol, Seoul 08826 - South Korea

^[8] CIRAD, UMR ASTRE, F-34398 Montpellier - France

^[9] Univ Montpellier, ASTRE, CIRAD, INRA, Montpellier - France

^[10] James Cook Univ, Coll Publ Hlth Med & Vet Sci, Hlth Res Grp 1, Townsville, Qld 4811 - Australia

^[11] Agcy Populat Ecol & Nat Conservancy, Gerbrunn - Germany

^[12] CSIC, Museo Nacl Ciencias Nat, C Jose Gutierrez Abascal 2, E-28006 Madrid - Spain

^[13] Univ Guyane, CNRS, IFREMER, LEEISA, Cayenne 97300 - French Guiana

^[14] Univ Porto, InBIO, CIBIO Ctr Invest Biodiversidade & Recursos Genet, P-4485661 Vairao - Portugal

^[15] Royal Zool Soc Scotland, Conservat Programmes, Edinburgh, Midlothian - Scotland

^[16] Oklahoma State Univ, Dept Integrat Biol, Life Sci West 113, Stillwater, OK 74078 - USA

^[17] Nat Hist Museum, Life Sci, London SW7 5BD - England

^[18] Uppsala Univ, EBC, Dept Ecol & Genet, Norbyv 18D, SE-75236 Uppsala - Sweden

^[19] Univ Michigan, Dept Ecol & Evolutionary Biol, Ann Arbor, MI 48109 - USA

^[20] Univ Estadual Campinas, Inst Biol, Dept Biol Anim, Lab Hist Nat Anfibios Brasileiros, BR-13083862 Campinas, SP - Brazil

^[21] Endem Species Res Inst, Zool Div, 1 Ming Shen East Rd, Nantou 552 - Taiwan

^[22] UFZ Helmholtz Ctr Environm Res, Dept Conservat Biol, Permoserstr 15, D-04318 Leipzig - Germany

^[23] Univ Toulouse, CNRS, ECOLAB, INPT, UPS, Toulouse - France

^[24] Univ Ghent, Dept Pathol Bacteriol & Avian Dis, Fac Vet Med, Salisburylaan 133, B-9820 Merelbeke - Belgium

^[25] Univ Paul Valery Montpellier, Univ Montpellier, PSL Res Univ, CEFE, UMR 5175, CNRS, EPHE, Biogeog & Ecol Vertebres, Montpellier - France

^[26] Amphibian & Reptile Conservat ARC Trust, 655A Christchurch Rd, Bournemouth BH1 4AP, Dorset - England

^[27] Durrell Wildlife Conservat Trust, Madagascar Programme, Antananarivo - Madagascar

^[28] IUCN SSC Amphibian Specialist Grp Madagascar, Antananarivo 101 - Madagascar

^[29] Univ Zurich, Dept Evolutionary Biol & Environm Studies, Winterthurerstr 190, CH-8057 Zurich - Switzerland

^[30] Univ Neuchatel, Info Fauna Karch, Bellevaux 51, UniMail Batiment 6, CH-2000 Neuchatel - Switzerland

^[31] APHA, Natl Wildlife Management Ctr, Woodchester Pk GL10 3UJ, Glos - England

^[32] Nonprofit Assoc Zirichiltaggi Sardinia Wildlife C, Str Vicinale Filigheddu 62-C, I-07100 Sassari - Italy

^[33] ONG Ranita Darwin, Nataniel Cox 152, Santiago - Chile

^[34] Hungarian Nat Hist Museum, Dept Zool, Collect Amphibians & Reptiles, Baross U 13, H-1088 Budapest - Hungary

^[35] Cornell Univ, Dept Ecol & Evolutionary Biol, Ithaca, NY 14853 - USA

^[36] Univ Maine, Sch Biol & Ecol, Orono, ME 04469 - USA

• Document type: Journal article
• Source: SCIENTIFIC REPORTS; v. 8, MAY 17 2018.
• Web of Science Citations: 8

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. (AU)

• FAPESP's process: 16/25358-3 - The chytrid fungus: from its origins to its consequences
• Grantee: Luis Felipe de Toledo Ramos Pereira
• Support Opportunities: Research Projects - Thematic Grants