Abstract

Viral erythrocytic necrosis (VEN) affects over 20 species of marine and anadromous fishes in the North Atlantic and North Pacific Oceans. However, the distribution and strain variation of its viral causative agent, erythrocytic necrosis virus (ENV), has not been well characterized within Pacific salmon. Here, metatranscriptomic sequencing of Chinook salmon revealed that ENV infecting salmon was closely related to ENV from Pacific herring, with inferred amino-acid sequences from Chinook salmon being 99% identical to those reported for herring. Sequence analysis also revealed 89 protein-encoding sequences attributed to ENV, greatly expanding the amount of genetic information available for this virus. High-throughput PCR of over 19,000 fish showed that ENV is widely distributed in the NE Pacific Ocean and was detected in 12 of 16 tested species, including in 27% of herring, 38% of anchovy, 17% of pollock, and 13% of sand lance. Despite frequent detection in marine fish, ENV prevalence was significantly lower in fish from freshwater (0.03%), as assessed with a generalized linear mixed effects model (p = 5.5 × 10−8). Thus, marine fish are likely a reservoir for the virus. High genetic similarity between ENV obtained from salmon and herring also suggests that transmission between these hosts is likely.

Highlights

  • Viral erythrocytic necrosis (VEN) is a disease associated with severe blood abnormalities in infected fish which has caused mass mortality in Pacific herring (Clupea pallasii) [1]

  • Metatranscriptomic sequencing of Chinook salmon with high erythrocytic necrosis virus (ENV) loads revealed high sequence identity between ENV sequences found in Chinook salmon and viral sequences from GenBank associated with Pacific herring

  • Nucleotide variation of ENV from herring and Chinook salmon was low, with ENV sequences from Pacific herring [2,4] having >99% nucleotide identity with the sequence obtained from Chinook salmon in this study, indicating that both viruses belong within the same genus, and likely the same species

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Summary

Introduction

Viral erythrocytic necrosis (VEN) is a disease associated with severe blood abnormalities in infected fish which has caused mass mortality in Pacific herring (Clupea pallasii) [1]. The disease is traditionally diagnosed by microscopic examination of stained blood smears for the presence of inclusion bodies within the cytoplasm of infected erythrocytes. Viruses 2019, 11, 358 infected erythrocytes contained icosahedral virions, which were named erythrocytic necrosis virus (ENV) [2]. Based on the small amount of available ENV genomic sequence, the virus has been assigned to a new putative genus within the Iridoviridae (a family of double-stranded DNA viruses), comprising other erythrocytic viruses from ectothermic hosts [2,4]. Further genomic sequencing of ENV may provide a reference for studying genetic variation geographically, and among fish species. The only verified ENV sequences available in GenBank encode ATPase, the major capsid protein (MCP), DNA-dependent DNA polymerase, and DNA-dependent RNA polymerase

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