title: Isolation of salmon pancreas disease virus (SPDV) in cell culture and its ability to protect against infection by the 'wild-type' agent.
abstract: A Scottish salmon pancreas disease virus (SPDV) has been isolated and its optimum growth conditions determined. Although several fish cell lines have been tested, successful culture was achieved only with CHSE-214 cells. Cytopathic effects were observed after 5 days. The highest virus titres, calculated by microtitration assay, were reached at 15 degrees C. After 7-9 days post-inoculation, CHSE-214 cell supernatants contained between 10(7)-10(5) TCID50 ml(-1) The cultured isolate is chloroform- and pH 3.0-sensitive, and virions are 50-60 nm in diameter. These characteristics are similar to the Irish SPDV isolates. The culture isolate induced typical pancreas disease (PD) lesions in experimentally infected Atlantic salmon and convalescent fish were resistant to experimental infection with PD-infective kidney homogenates obtained by serial in vivo passages from a PD-infected farmed salmon (termed wild-type SPDV). Furthermore, fish immunised with the inactivated cultured virus were protected against a cohabitation challenge with the wild-type virus. Immunised fish sera showed virus-neutralising activity before challenge (7 weeks post-immunisation) and from 3-6 weeks post-challenge, when sera from non-immunised fish did not neutralise the virus. At 6 weeks post-cohabitation challenge, previously immunised fish had neutralising titres of up to 1:65. Following intraperitoneal (i.p.) challenge, immunised fish showed neutralising titres as high as 1:226 at 8 weeks post-challenge. Non-immunised fish injected i.p. with the wild-type virus developed serum-neutralising activity against the cultured isolate when sampled 8 weeks after infection, confirming an antigenic relationship between the wild-type and cultured virus. The results demonstrate that the tissue culture-adapted isolate of SPDV could be successfully used to protect against challenge by the wild-type virus and could therefore have potential use as an inactivated vaccine against PD.
authors.affiliation: Marine Laboratory Aberdeen, Scotland, UK.
authors.forename: M V
authors.initials: MV
authors.lastname: López-Dóriga
authors.forename: D A
authors.initials: DA
authors.lastname: Smail
authors.forename: R J
authors.initials: RJ
authors.lastname: Smith
authors.forename: A
authors.initials: A
authors.lastname: Doménech
authors.forename: J
authors.initials: J
authors.lastname: Castric
authors.forename: P D
authors.initials: PD
authors.lastname: Smith
authors.forename: A E
authors.initials: AE
authors.lastname: Ellis
mesh_terms: D000524:Alphavirus / Q000276:immunology / Q000302:isolation & purification*; D018354:Alphavirus Infections / Q000276:immunology / Q000662:veterinary* / Q000821:virology; D000818:Animals; D002478:Cells, Cultured; D005393:Fish Diseases / Q000276:immunology* / Q000821:virology; D006863:Hydrogen-Ion Concentration; D007274:Injections, Intraperitoneal / Q000662:veterinary; D009500:Neutralization Tests / Q000662:veterinary; D017686:Oncorhynchus mykiss; D010179:Pancreas / Q000473:pathology; D010182:Pancreatic Diseases / Q000276:immunology / Q000662:veterinary* / Q000821:virology; D020317:Salmo salar; D012487:Salmonidae; D013696:Temperature; D015164:Vaccines, Inactivated; D019562:Viral Load / Q000662:veterinary
chemical_list: D015164:Vaccines, Inactivated