Development and worldwide use of non-lethal, and minimal population-level impact, protocols for the isolation of amphibian chytrid fungi.
Matthew C FisherPria GhoshJennifer M G SheltonKieran BatesLola BrookesClaudia WierzbickiGonçalo M RosaRhys A FarrerDavid M AanensenMario Alvarado-RybakArnaud BatailleLee BergerSusanne BöllJaime BoschFrances C ClareElodie A CourtoisAngelica CrottiniAndrew A CunninghamThomas M Doherty-BoneFikirte GebresenbetDavid J GowerJacob HöglundTimothy Y JamesThomas S JenkinsonTiffany A KoschCarolina LambertiniAnssi LaurilaChun-Fu LinAdeline LoyauAn MartelSara MeurlingClaude MiaudPete MintingSerge NdriantsoaSimon J O'HanlonFrank PasmansTsanta RakotonanaharyFalitiana C E RabemananjaraLuisa P RibeiroDirk S SchmellerBenedikt R SchmidtLee SkerrattFreya SmithClaudio Soto-AzatGiulia TessaLuís Felipe ToledoAndrés Valenzuela-SánchezRuhan VersterJudit VörösBruce WaldmanRebecca J WebbChe WeldonEmma WombwellKelly R ZamudioJoyce E LongcoreTrenton W J GarnerPublished in: Scientific reports (2018)
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.