Isolation and characterization of the fall Chinook aquareovirus

Virol J. 2017 Sep 5;14(1):170. doi: 10.1186/s12985-017-0839-9.

Abstract

Background: Salmon are paramount to the economy, ecology, and history of the Pacific Northwest. Viruses constitute one of the major threats to salmon health and well-being, with more than twenty known virus species that infect salmon. Here, we describe the isolation and characterization of the fall Chinook aquareovirus, a divergent member of the species Aquareovirus B within the family Reoviridae.

Methods: The virus was first found in 2014 as part of a routine adult broodstock screening program in which kidney and spleen tissue samples from healthy-appearing, adult fall Chinook salmon (Oncorhynchus tshawytscha) returning to a hatchery in Washington State produced cytopathic effects when inoculated onto a Chinook salmon embryo cell line (CHSE-214). The virus was not able to be confirmed by an RT-PCR assay using existing aquareovirus pan-species primers, and instead was identified by metagenomic next-generation sequencing. Metagenomic next-generation sequencing was used to recover the full genome and completed using 3' RACE.

Results: The genome of the fall Chinook aquareovirus contains 11 segments of double-stranded RNA totaling 23.3 kb, with each segment flanked by the canonical sequence termini found in the aquareoviruses. Sequence comparisons and a phylogenetic analysis revealed a nucleotide identity of 63.2% in the VP7 gene with the Green River Chinook virus, placing the new isolate in the species Aquareovirus B. A qRT-PCR assay was developed targeting the VP2, which showed rapid growth of the isolate during the initial 5 days in culture using CHSE-214 cells.

Conclusions: This sequence represents the first complete genome of an Aquareovirus B species. Future studies will be required to understand the potential pathogenicity and epidemiology of the fall Chinook aquareovirus.

Keywords: Aquareovirus; Aquareovirus B; Chinook; Fall chinook; Salmon virus.

MeSH terms

  • Animals
  • Antigens, Viral / chemistry
  • Antigens, Viral / genetics
  • Cell Line
  • Fish Diseases / pathology
  • Fish Diseases / virology*
  • Genome, Viral*
  • Metagenomics
  • Phylogeny
  • RNA, Double-Stranded / genetics
  • RNA, Viral / genetics*
  • Real-Time Polymerase Chain Reaction
  • Reoviridae / classification
  • Reoviridae / genetics*
  • Reoviridae / growth & development
  • Reoviridae / isolation & purification*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Salmon / virology*
  • Sequence Homology, Amino Acid
  • Sequence Homology, Nucleic Acid

Substances

  • Antigens, Viral
  • RNA, Double-Stranded
  • RNA, Viral