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      Chytridiomycosis Outbreak in a Chilean Giant Frog ( Calyptocephalella gayi) Captive Breeding Program: Genomic Characterization and Pathological Findings

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          Abstract

          Emerging infectious diseases in wildlife are increasingly associated with animal mortality and species declines, but their source and genetic characterization often remains elusive. Amphibian chytridiomycosis, caused by the fungus Batrachochytrium dendrobatidis ( Bd), has been associated with catastrophic and well-documented amphibian population declines and extinctions at the global scale. We used histology and whole-genome sequencing to describe the lesions caused by, and the genetic variability of, two Bd isolates obtained from a mass mortality event in a captive population of the threatened Chilean giant frog ( Calyptocephalella gayi). This was the first time an association between Bd and high mortality had been detected in this charismatic and declining frog species. Pathological examinations revealed that 30 dead metamorphosed frogs presented agnathia or brachygnathia, a condition that is reported for the first time in association with chytridiomycosis. Phylogenomic analyses revealed that Bd isolates (PA1 and PA2) from captive C. gayi group with other Bd isolates (AVS2, AVS4, and AVS7) forming a single highly supported Chilean Bd clade within the global panzootic lineage of Bd ( BdGPL). These findings are important to inform the strengthening of biosecurity measures to prevent the impacts of chytridiomycosis in captive breeding programs elsewhere.

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          The Sequence Alignment/Map format and SAMtools

          Summary: The Sequence Alignment/Map (SAM) format is a generic alignment format for storing read alignments against reference sequences, supporting short and long reads (up to 128 Mbp) produced by different sequencing platforms. It is flexible in style, compact in size, efficient in random access and is the format in which alignments from the 1000 Genomes Project are released. SAMtools implements various utilities for post-processing alignments in the SAM format, such as indexing, variant caller and alignment viewer, and thus provides universal tools for processing read alignments. Availability: http://samtools.sourceforge.net Contact: rd@sanger.ac.uk
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            Fast and accurate short read alignment with Burrows–Wheeler transform

            Motivation: The enormous amount of short reads generated by the new DNA sequencing technologies call for the development of fast and accurate read alignment programs. A first generation of hash table-based methods has been developed, including MAQ, which is accurate, feature rich and fast enough to align short reads from a single individual. However, MAQ does not support gapped alignment for single-end reads, which makes it unsuitable for alignment of longer reads where indels may occur frequently. The speed of MAQ is also a concern when the alignment is scaled up to the resequencing of hundreds of individuals. Results: We implemented Burrows-Wheeler Alignment tool (BWA), a new read alignment package that is based on backward search with Burrows–Wheeler Transform (BWT), to efficiently align short sequencing reads against a large reference sequence such as the human genome, allowing mismatches and gaps. BWA supports both base space reads, e.g. from Illumina sequencing machines, and color space reads from AB SOLiD machines. Evaluations on both simulated and real data suggest that BWA is ∼10–20× faster than MAQ, while achieving similar accuracy. In addition, BWA outputs alignment in the new standard SAM (Sequence Alignment/Map) format. Variant calling and other downstream analyses after the alignment can be achieved with the open source SAMtools software package. Availability: http://maq.sourceforge.net Contact: rd@sanger.ac.uk
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              Cutadapt removes adapter sequences from high-throughput sequencing reads

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                Author and article information

                Contributors
                Journal
                Front Vet Sci
                Front Vet Sci
                Front. Vet. Sci.
                Frontiers in Veterinary Science
                Frontiers Media S.A.
                2297-1769
                24 September 2021
                2021
                : 8
                : 733357
                Affiliations
                [1] 1Sustainability Research Centre & PhD Program in Conservation Medicine, Faculty of Life Sciences, Universidad Andres Bello , Santiago, Chile
                [2] 2Institute of Zoology, Zoological Society of London , London, United Kingdom
                [3] 3Núcleo de Ciencias Aplicadas en Ciencias Veterinarias y Agronómicas, Universidad de las Américas , Santiago, Chile
                [4] 4Criadero y Centro de Exhibición de la Rana Chilena Calyptocephalella gayi , Santiago, Chile
                [5] 5Department of Infectious Disease Epidemiology, MRC Centre for Global Infectious Disease Analysis, School of Public Health, Imperial College London , London, United Kingdom
                [6] 6ONG Ranita de Darwin , Valdivia, Chile
                [7] 7Instituto de Conservación, Biodiversidad y Territorio, Facultad de Ciencias Forestales y Recursos Naturales, Universidad Austral de Chile , Valdivia, Chile
                Author notes

                Edited by: Amanda Linda Jean Duffus, Gordon State College, United States

                Reviewed by: Rebecca Hardman, The University of Tennessee, Knoxville, United States; An Martel, Ghent University, Belgium

                *Correspondence: Mario Alvarado-Rybak maalry@ 123456gmail.com

                This article was submitted to Veterinary Infectious Diseases, a section of the journal Frontiers in Veterinary Science

                Article
                10.3389/fvets.2021.733357
                8497818
                34631859
                5a83de9c-31a0-4b5f-b462-7316c7de398a
                Copyright © 2021 Alvarado-Rybak, Acuña, Peñafiel-Ricaurte, Sewell, O'Hanlon, Fisher, Valenzuela-Sánchez, Cunningham and Azat.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 30 June 2021
                : 27 August 2021
                Page count
                Figures: 4, Tables: 0, Equations: 0, References: 68, Pages: 9, Words: 6610
                Funding
                Funded by: Fondo Nacional de Desarrollo Científico y Tecnológico, doi 10.13039/501100002850;
                Categories
                Veterinary Science
                Original Research

                agnathia,amphibians,batrachochytrium dendrobatidis,bdgpl,brachygnathia,chile,emerging infectious disease,whole-genome sequencing

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