title: Natural infection of Atlantic salmon (Salmo salar L.) with salmonid alphavirus 3 generates numerous viral deletion mutants.
abstract: Salmon pancreas disease virus (SPDV) also referred to as salmonid alphavirus (SAV) is a virus causing pancreas disease in Atlantic salmon (Salmo salar L.) and rainbow trout (Oncorhynchus mykiss). Although the virus causes an economically important disease, relatively few full-length genome sequences of SAV strains are currently available. Here, we report full-length genome sequences of nine SAV3 strains from sites farming Atlantic salmon geographically spread along the Norwegian coastline. The virus genomes were sequenced directly from infected heart tissue, to avoid culture selection bias. Sequence analysis confirmed a high level of sequence identity within SAV3 strains, with a mean nucleotide diversity of 0.11 %. Sequence divergence was highest in 6K and E2, while lowest in the capsid protein and the non-structural proteins (nsP4 and nsP2). This study reports for the first time that numerous defective viruses containing genome deletions are generated during natural infection with SAV. Deletions occurred in all virus strains and were not distributed randomly throughout the genome but instead tended to aggregate in certain areas. We suggest imprecise homologous recombination as an explanation for generation of defective viruses with genome deletions. The presence of such viruses, provides a possible explanation for the difficulties in isolating SAV in cell culture. Primary virus isolation was successfully achieved for only two of eight strains, despite extensive attempts using three different cell lines. Both SAV isolates were easily propagated further and concomitant viral deletion mutants present in clinically infected heart tissue were maintained following serial passage in CHH-1 cells.
authors.affiliation: Norwegian School of Veterinary Science, Department of Basic Sciences and Aquatic Medicine, P.O. Box 8146 Dep., 0033 Oslo Norway.
authors.forename: Elin
authors.initials: E
authors.lastname: Petterson
authors.affiliation: Norwegian School of Veterinary Science, Department of Basic Sciences and Aquatic Medicine, P.O. Box 8146 Dep., 0033 Oslo Norway.
authors.forename: Marit
authors.initials: M
authors.lastname: Stormoen
authors.affiliation: Norwegian School of Veterinary Science, Department of Basic Sciences and Aquatic Medicine, P.O. Box 8146 Dep., 0033 Oslo Norway.
authors.forename: Øystein
authors.initials: Ø
authors.lastname: Evensen
authors.affiliation: Norwegian School of Veterinary Science, Department of Basic Sciences and Aquatic Medicine, P.O. Box 8146 Dep., 0033 Oslo Norway.
authors.forename: Aase B
authors.initials: AB
authors.lastname: Mikalsen
authors.affiliation: Norwegian School of Veterinary Science, Department of Basic Sciences and Aquatic Medicine, P.O. Box 8146 Dep., 0033 Oslo Norway.
authors.forename: Øyvind
authors.initials: Ø
authors.lastname: Haugland
mesh_terms: D000524:Alphavirus / Q000235:genetics* / Q000302:isolation & purification; D018354:Alphavirus Infections / Q000662:veterinary* / Q000821:virology; D000818:Animals; D005393:Fish Diseases / Q000821:virology*; D014644:Genetic Variation; D016679:Genome, Viral; D006321:Heart / Q000821:virology; D008969:Molecular Sequence Data; D009664:Norway; D012367:RNA, Viral / Q000235:genetics; D020317:Salmo salar / Q000821:virology*; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion
chemical_list: D012367:RNA, Viral