Identification of a piscine reovirus-related pathogen in proliferative darkening syndrome (PDS) infected brown trout (Salmo trutta fario) using a next-generation technology detection pipeline

Autor: Kuehn, Ralph, Stoeckle, Bernhard C., Young, Marc, Popp, Lisa, Taeubert, Jens-Eike, Pfaffl, Michael W., Geist, Juergen
Jazyk: angličtina
Rok vydání: 2018
Předmět:
Next-Generation Sequencing
Trout
Bioinformatics
Molecular biology
Microarrays
Immunology
lcsh:Medicine
Sequence Databases
Gene Expression
Research and Analysis Methods
Pathology and Laboratory Medicine
Database and Informatics Methods
Fish Diseases
Sequencing techniques
Species Specificity
Medicine and Health Sciences
Genetics
Animals
DNA sequencing
lcsh:Science
Immune Response
Phylogeny
Oligonucleotide Array Sequence Analysis
Sequence Analysis
RNA
Gene Expression Profiling
lcsh:R
Organisms
Biology and Life Sciences
Eukaryota
Computational Biology
High-Throughput Nucleotide Sequencing
Genomics
Genome Analysis
Europe
Orthoreovirus
Fish
Biological Databases
Molecular biology techniques
Bioassays and Physiological Analysis
Liver
Osteichthyes
Vertebrates
RNA
Viral
lcsh:Q
Pathogens
Sequence Analysis
Transcriptome Analysis
Research Article
Zdroj: PLoS ONE
PLoS ONE, Vol 13, Iss 10, p e0206164 (2018)
ISSN: 1932-6203
Popis: The proliferative darkening syndrome (PDS) is an annually recurring disease that causes species-specific die-off of brown trout (Salmo trutta fario) with a mortality rate of near 100% in pre-alpine rivers of central Europe. So far the etiology and causation of this disease is still unclear. The objective of this study was to identify the cause of PDS using a next-generation technology detection pipeline. Following the hypothesis that PDS is caused by an infectious agent, brown trout specimens were exposed to water from a heavily affected pre-alpine river with annual occurrence of the disease. Specimens were sampled over the entire time period from potential infection through death. Transcriptomic analysis (microarray) and RT-qPCR of brown trout liver tissue evidenced strong gene expression response of immune-associated genes. Messenger RNA of specimens with synchronous immune expression profiles were ultra-deep sequenced using next-generation sequencing technology (NGS). Bioinformatic processing of generated reads and gap-filling Sanger re-sequencing of the identified pathogen genome revealed strong evidence that a piscine-related reovirus is the causative organism of PDS. The identified pathogen is phylogenetically closely related to the family of piscine reoviruses (PRV) which are considered as the causation of different fish diseases in Atlantic and Pacific salmonid species such as Salmo salar and Onchorhynchus kisutch. This study also highlights that the approach of first screening immune responses along a timeline in order to identify synchronously affected stages in different specimens which subsequently were ultra-deep sequenced is an effective approach in pathogen detection. In particular, the identification of specimens with synchronous molecular immune response patterns combined with NGS sequencing and gap-filling re-sequencing resulted in the successful pathogen detection of PDS.
Databáze: OpenAIRE
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