Infection experiments with novel Piscine orthoreovirus from rainbow trout (Oncorhynchus mykiss) in salmonids

PLoS One. 2017 Jul 5;12(7):e0180293. doi: 10.1371/journal.pone.0180293. eCollection 2017.

Abstract

A new disease in farmed rainbow trout (Onchorhyncus mykiss) was described in Norway in 2013. The disease mainly affected the heart and resembled heart and skeletal muscle inflammation (HSMI) in Atlantic salmon (Salmo salar L.). HSMI is associated with Piscine orthoreovirus (PRV), and a search for a similar virus in the diseased rainbow trout led to detection of a sequence with 85% similarity to PRV. This finding called for a targeted effort to assess the risk the new PRV-variant pose on farmed rainbow trout and Atlantic salmon by studying infection and disease pathogenesis, aiming to provide more diagnostic knowledge. Based on the genetic relationship to PRV, the novel virus is referred to as PRV-Oncorhynchus mykiss (PRV-Om) in contrast to PRV-Salmo salar (PRV-Ss). In experimental trials, intraperitoneally injected PRV-Om was shown to replicate in blood in both salmonid species, but more effectively in rainbow trout. In rainbow trout, the virus levels peaked in blood and heart of cohabitants 6 weeks post challenge, along with increased expression of antiviral genes (Mx and viperin) in the spleen, with 80-100% of the cohabitants infected. Heart inflammation was diagnosed in all cohabitants examined 8 weeks post challenge. In contrast, less than 50% of the Atlantic salmon cohabitants were infected between 8 and 16 weeks post challenge and the antiviral response in these fish was very low. From 12 weeks post challenge and onwards, mild focal myocarditis was demonstrated in a few virus-positive salmon. In conclusion, PRV-Om infects both salmonid species, but faster transmission, more notable antiviral response and more prominent heart pathology were observed in rainbow trout.

MeSH terms

  • Animals
  • Denmark
  • Fish Diseases / diagnosis
  • Fish Diseases / transmission
  • Fish Diseases / virology*
  • Fish Proteins / blood
  • Fish Proteins / genetics
  • Gene Expression
  • Heart / virology
  • Hemoglobins / analysis
  • Host-Pathogen Interactions
  • Muscle, Skeletal / virology
  • Norway
  • Oncorhynchus mykiss / blood
  • Oncorhynchus mykiss / genetics
  • Oncorhynchus mykiss / virology*
  • Orthoreovirus / genetics
  • Orthoreovirus / pathogenicity
  • Orthoreovirus / physiology*
  • RNA, Viral / genetics
  • Reoviridae Infections / diagnosis
  • Reoviridae Infections / transmission
  • Reoviridae Infections / virology*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Salmo salar / blood
  • Salmo salar / genetics
  • Salmo salar / virology*
  • Virulence

Substances

  • Fish Proteins
  • Hemoglobins
  • RNA, Viral

Grants and funding

The pilot study (trial 1), the Atlantic salmon main study (trial 3), and histopathological analyses for both species were funded by the Norwegian Veterinary Institute (NVI). The rainbow trout study (trial 2) was funded by the EU Reference Laboratory for Fish Diseases, DTU National Veterinary Institute and FAPESP (Sao Paulo Research Foundation) Research internship abroad (2015/04027-6). Experiments performed at the Norwegian University of Life Sciences were funded by the Norwegian Research Council grant # 237315/E40. Many thanks to Renate Johansen and Monika Hjortaas (NVI) for fruitful initial discussions on the pilot study design, and to Even Thoen (NVI) for practical assistance setting up the pilot study. The Atlantic salmon trial at VESO Vikan was managed by Anne Ramstad and crew. We greatly appreciate the help from several NVI and DTU Vet technicians. In particular; Elisabeth Dahl Nybø (NVI) for sample handling in the Atlantic salmon main study and RNA preparation for immune analysis, Randi Faller (NVI) for RT-qPCR analysis of immune genes, Linh Tran (NVI) for immunohistochemistry, Didde Hedegaard Sørensen and Troels Secher Rundqvist (DTU Vet) for nucleic acid purification and RT-qPCR and Kari Karbech Mouritsen (DTU Vet) for sample handling and preparation.