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      Discriminating lineages of Batrachochytrium dendrobatidis using quantitative PCR

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          Abstract

          The ability to detect and monitor infectious disease in a phylogenetically informative manner is critical for their management. Phylogenetically informative diagnostic tests enable patterns of pathogen introduction or changes in the distribution of genotypes to be measured, enabling research into the ecology of the pathogen. Batrachochytrium dendrobatidis ( Bd), a causative agent of chytridiomycosis in amphibian populations, emerged worldwide in the 21st century and is composed of six lineages which are display varying levels of virulence in their hosts. Research into the distribution, ecology and pathogenicity of these lineages has been hampered by an inability to type lineage efficiently. Here, we describe a lineage‐specific TaqMan qPCR assay that differentiates the two lineages of Bd most commonly associated with chytridiomycosis: BdGPL and BdCAPE. We demonstrate how this assay can be used for the surveillance of wild populations of amphibians in Southern Africa using skin swabs, tissue samples and cultured isolates.

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          see also the Perspective by Claudio Azat

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          Most cited references26

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          Nuclear ribosomal internal transcribed spacer (ITS) region as a universal DNA barcode marker for Fungi.

          Six DNA regions were evaluated as potential DNA barcodes for Fungi, the second largest kingdom of eukaryotic life, by a multinational, multilaboratory consortium. The region of the mitochondrial cytochrome c oxidase subunit 1 used as the animal barcode was excluded as a potential marker, because it is difficult to amplify in fungi, often includes large introns, and can be insufficiently variable. Three subunits from the nuclear ribosomal RNA cistron were compared together with regions of three representative protein-coding genes (largest subunit of RNA polymerase II, second largest subunit of RNA polymerase II, and minichromosome maintenance protein). Although the protein-coding gene regions often had a higher percent of correct identification compared with ribosomal markers, low PCR amplification and sequencing success eliminated them as candidates for a universal fungal barcode. Among the regions of the ribosomal cistron, the internal transcribed spacer (ITS) region has the highest probability of successful identification for the broadest range of fungi, with the most clearly defined barcode gap between inter- and intraspecific variation. The nuclear ribosomal large subunit, a popular phylogenetic marker in certain groups, had superior species resolution in some taxonomic groups, such as the early diverging lineages and the ascomycete yeasts, but was otherwise slightly inferior to the ITS. The nuclear ribosomal small subunit has poor species-level resolution in fungi. ITS will be formally proposed for adoption as the primary fungal barcode marker to the Consortium for the Barcode of Life, with the possibility that supplementary barcodes may be developed for particular narrowly circumscribed taxonomic groups.
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            Amphibian fungal panzootic causes catastrophic and ongoing loss of biodiversity

            Anthropogenic trade and development have broken down dispersal barriers, facilitating the spread of diseases that threaten Earth’s biodiversity. We present a global, quantitative assessment of the amphibian chytridiomycosis panzootic, one of the most impactful examples of disease spread, and demonstrate its role in the decline of at least 501 amphibian species over the past half-century, including 90 presumed extinctions. The effects of chytridiomycosis have been greatest in large-bodied, range-restricted anurans in wet climates in the Americas and Australia. Declines peaked in the 1980s, and only 12% of declined species show signs of recovery, whereas 39% are experiencing ongoing decline. There is risk of further chytridiomycosis outbreaks in new areas. The chytridiomycosis panzootic represents the greatest recorded loss of biodiversity attributable to a disease.
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              Batrachochytrium Dendrobatidis gen. et sp. nov., a Chytrid Pathogenic to Amphibians

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                Author and article information

                Contributors
                pria.ghosh@gmail.com
                ruhan.verster@gmail.com
                t.sewell@imperial.ac.uk
                simon.ohanlon@imperial.ac.uk
                Lola.Brookes@ioz.ac.uk
                adrien.rieux@cirad.fr
                Trent.Garner@ioz.ac.uk
                che.weldon@nwu.ac.za
                matthew.fisher@imperial.ac.uk
                Journal
                Mol Ecol Resour
                Mol Ecol Resour
                10.1111/(ISSN)1755-0998
                MEN
                Molecular Ecology Resources
                John Wiley and Sons Inc. (Hoboken )
                1755-098X
                1755-0998
                12 February 2021
                July 2021
                : 21
                : 5 ( doiID: 10.1111/men.v21.5 )
                : 1452-1459
                Affiliations
                [ 1 ] MRC Centre for Global Infectious Disease Analysis Department of Infectious Disease Epidemiology Imperial College London London UK
                [ 2 ] Unit for Environmental Sciences and Management North‐West University Potchefstroom South Africa
                [ 3 ] Institute of Zoology Zoological Society of London London UK
                [ 4 ] The Royal Veterinary College Hatfield UK
                [ 5 ] CIRAD, UMR, PVBMT St Pierre La Réunion France
                Author notes
                [*] [* ] Correspondence

                Pria N. Ghosh, MRC Centre for Global Infectious Disease Analysis, Department of Infectious Disease Epidemiology, Imperial College London W2 1PG, London, UK.

                Email: pria.ghosh@ 123456gmail.com

                Author information
                https://orcid.org/0000-0002-9668-6662
                https://orcid.org/0000-0003-2974-1155
                https://orcid.org/0000-0002-7239-4817
                https://orcid.org/0000-0002-6852-3911
                https://orcid.org/0000-0001-5354-0244
                https://orcid.org/0000-0002-7221-0010
                https://orcid.org/0000-0002-0336-9706
                https://orcid.org/0000-0001-6716-0787
                https://orcid.org/0000-0002-1862-6402
                Article
                MEN13299
                10.1111/1755-0998.13299
                8651002
                33232563
                5ea169e6-d5dd-4df6-906f-aa2b96795c38
                © 2020 The Authors. Molecular Ecology Resources published by John Wiley & Sons Ltd

                This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 03 November 2020
                : 15 August 2020
                : 09 November 2020
                Page count
                Figures: 4, Tables: 0, Pages: 8, Words: 4571
                Funding
                Funded by: Morris Animal Foundation , doi 10.13039/100001250;
                Funded by: Leverhulme Trust , doi 10.13039/501100000275;
                Award ID: RPG‐2014‐273
                Funded by: Natural Environment Research Council , doi 10.13039/501100000270;
                Award ID: NERC NE/E006841/1
                Categories
                Resource Article
                RESOURCE ARTICLES
                Molecular and Statistical Advances
                Custom metadata
                2.0
                July 2021
                Converter:WILEY_ML3GV2_TO_JATSPMC version:6.0.9 mode:remove_FC converted:07.12.2021

                Ecology
                batrachochytrium dendrobatidis,chytrid,diagnostic,lineage,qpcr
                Ecology
                batrachochytrium dendrobatidis, chytrid, diagnostic, lineage, qpcr

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