title: In vitro adaptation of SAV3 in cell culture correlates with reduced in vivo replication capacity and virulence to Atlantic salmon (Salmo salar L.) parr.
abstract: Salmonid alphavirus (SAV) is the causative agent of pancreas disease affecting Atlantic salmon and rainbow trout and causes a major burden to the aquaculture industry. This study describes a Norwegian subtype SAV3 virus isolate (SAV3-H10) subjected to serial passages in Chinook salmon embryo cells (CHSE-214) followed by Asian Grouper skin cells (AGK). Two passages from CHSE and one after transfer to AGK cells were chosen for further investigation, based on variation in degree and development of cytopathic effect (CPE). After plaque purification, several in vitro studies were performed. Cell viability after infection, viral replication and ability to cause morphological changes in CHSE and AGK cells was studied for the three isolates. The AGK-transferred isolate was identified with the strongest abilities to reduce cell viability, replicate more and cause more CPE in cell culture when compared with the early and late CHSE-grown isolates. Subsequently, the isolates were tested in an experimental fish challenge, showing higher viral load and higher pathological score for the least cell-cultured isolate. Full-length sequencing of the viral genome of the three isolates revealed divergence in four amino acid positions and the AGK-grown isolate also had a 3 nt deletion in the 3'UTR. In conclusion, we show that cell culture of SAV3-H10 selects for strains inducing earlier CPE in vitro with increased viral replication. In vivo, the effect is reversed, with lower replication levels and lower pathology scores in target organs. This study outlines a path to identify potential virulence motifs of SAV3.
authors: Norwegian University of Life Sciences, Department of Basic Science and Aquatic Medicine, PO Box 8146 Dep., 0033 Oslo, Norway.
authors: Elin
authors: E
authors: Petterson
authors: Norwegian University of Life Sciences, Department of Basic Science and Aquatic Medicine, PO Box 8146 Dep., 0033 Oslo, Norway.
authors: Tz-Chun
authors: TC
authors: Guo
authors: Norwegian University of Life Sciences, Department of Basic Science and Aquatic Medicine, PO Box 8146 Dep., 0033 Oslo, Norway.
authors: Øystein
authors: Ø
authors: Evensen
authors: Norwegian University of Life Sciences, Department of Basic Science and Aquatic Medicine, PO Box 8146 Dep., 0033 Oslo, Norway.
authors: Øyvind
authors: Ø
authors: Haugland
authors: Norwegian University of Life Sciences, Department of Basic Science and Aquatic Medicine, PO Box 8146 Dep., 0033 Oslo, Norway.
authors: Aase B
authors: AB
authors: Mikalsen
mesh_terms: D000220:Adaptation, Biological; D000524:Alphavirus / Q000235:genetics* / Q000254:growth & development* / Q000472:pathogenicity / Q000502:physiology; D000818:Animals; D002470:Cell Survival; D003588:Cytopathogenic Effect, Viral; D004847:Epithelial Cells / Q000166:cytology / Q000502:physiology / Q000821:virology; D050296:Microbial Viability; D020317:Salmo salar / Q000821:virology*; D012692:Serial Passage; D014774:Virulence; D014776:Virus Cultivation; D014779:Virus Replication