Sammendrag
Introduction:
Heart- and skeletal muscle inflammation (HSMI) is an emerging disease in Atlantic salmon (Salmo salar) farming. HSMI is associated with Piscine orthoreovirus (PRV) infection, but PRV is ubiquitous in farmed Atlantic salmon and thus present also in apparently healthy individuals. This has brought speculations if additional etiological factors are required, and experiments focusing on the causal relationship between PRV and HSMI are highly warranted.
Methodology:
A major bottleneck in PRV research has been the lack of cell lines that allow propagation of the virus. To bypass this, we propagated PRV in salmon, bled the fish at the peak of the infection, and purified virus particles from blood cells by gradient centrifugation. Electron microscopy, western blot and high-throughput sequencing all verified the purity of the viral particles. Two doses of purified PRV particles were inoculated into Atlantic salmon, which were cohabitated 50:50 with naïve individuals for eight weeks. PRV load and antiviral responses were examined by RT-qPCR, and histological analysis and immunostaining for PRV was performed in heart and muscle.
Results:
The purified virus replicated in inoculated fish, spread to naïve cohabitants, and induced histopathological changes consistent with HSMI. PRV specific staining was demonstrated in the pathological lesions. A dose-dependent response was observed; a high dose of virus gave earlier peak of the viral load and development of histopathological changes compared to a lower dose, but no difference in the severity of the disease.
Conclusions:
The experiment demonstrated that PRV can be purified from blood cells, and that PRV is the etiological agent of HSMI in Atlantic salmon.
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