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dc.contributor.advisorDel-Pozo, Jorgeen
dc.contributor.advisorCollet, Bertranden
dc.contributor.advisorDutia, Bernadetteen
dc.contributor.authorNoguera, Patricia Alinaen
dc.date.accessioned2018-11-20T11:39:02Z
dc.date.available2018-11-20T11:39:02Z
dc.date.issued2018-11-30
dc.identifier.urihttp://hdl.handle.net/1842/33257
dc.description.abstractSalmon Pancreas Disease Virus (SPDV) is the only viral species of the genus Alphavirus, family Togaviridae, affecting fish. SPDV induces two conditions historically recognised independently as Pancreas disease (PD) and Sleeping disease (SD), affecting Atlantic salmon (Salmo salar L) and rainbow trout (Oncorhynchus mykiss), respectively. Infection by SPDV can lead to clinical disease with characteristic acinar pancreatic necrosis and a range of myopathies of the skeletal and heart muscle. Mortality is not a necessary outcome of the disease and usually is not significant. However, affected fish stop eating and therefore present a reduced growth rate and the disease can also leave visible lesions at the fillet level that lead to downgrading at slaughter. SPDV can affect in the fresh and sea water environments, but a higher and most relevant impact reported in the latter. Historically, PD has posed a significant challenge to the Atlantic salmon farming industry in the UK, as well as in other salmon producing countries. This thesis was developed and conducted at Marine Scotland Science (MSS), the Scottish National Reference Laboratory, with the aim to contribute to knowledge gaps identified by the industry and research communities. The focus was on development and improvement of in vivo and in vitro infection models to assist with host pathogen interaction studies. In vivo work was to establish an experimental challenge model to induce SPDV infection in a more natural way than by intra-peritoneal (IP) injection. The first step involved selection of an infective SPDV isolate through a comparative IP challenge study. An infective isolate was then used to establish a co-habitation challenge model in “post smolts”, the sea-water stage predominantly affected by PD. Additionally, during this experiment assessment of viral tissue tropism along time and potential intra-subtype differences in infectivity was undertaken. In vitro work accounted for the more innovative part of this thesis with the development, optimization and application of an ex vivo cardiac primary culture originated from Atlantic salmon embryos. While fish origin aggregates of self-contracting cardiomyocytes had been previously isolated and suggested as a robust tool on human biomedical research and pharmacological and toxicology testing, paradoxically very little has been done to explore the approach of ex vivo primary cultures as a disease model with the specific goal for health issues affecting fish. The work involved an adaptation and refinement to produce salmon cardiac primary cultures (SCPCs). Once this was achieved, SCPCs could be kept under laboratory conditions with minimal maintenance for periods up to 6 months. Following this work, SCPCs were successfully challenged with different SPDV isolates as well as another cardiotropic viral agent (Infectious Salmon Anaemia, ISA). The kinetics of SPDV and ISA viral infection and one element of the immune response (i.e. expression of mx gene) were studied. As part of this study, the comparative response of SCPCs of diverse genetic backgrounds (i.e. IPN resistant vs. IPN sensitive) was also assessed. Differences were observed, which highlights potential usefulness of SCPCs to examine genotype-based differences in response to viral disease. Finally, SCPCs were used to examine the SPDV infection cycle ultrastructure by transmission electron microscopy (TEM). This work resulted in novel insights on the replication cycle of SPDV, drawing from the extensive literature in mammalian alphavirus work. With SPDV and other virus associated myocarditis severely affecting Atlantic salmon aquaculture at present, I believe that the SCPCs model represents the most relevant contribution of this PhD.en
dc.contributor.sponsorotheren
dc.language.isoen
dc.publisherThe University of Edinburghen
dc.relation.hasversionBruno DW, Noguera PA, Poppe TT. A Colour Atlas of Salmonid Diseases [Internet]. Dordrecht: Springer Netherlands; 2013. doi:10.1007/978-94-007-2010-7en
dc.relation.hasversionNoguera P, Collet B, Klinger M, Örün H, del Pozo J. Use of Salmon Cardiac Primary Cultures (SCPCs) of different genotypes for comparative kinetics of mx expression, viral load and ultrastructure pathology, after infection with Salmon Pancreas Disease Virus (SPDV). Fish Shellfish Immunol. 2018;72: 181–186. doi:10.1016/j.fsi.2017.10.059en
dc.relation.hasversionBruno D, Noguera P, Black J, Murray W, Macqueen D, Matejusova I. Identification of a wild reservoir of salmonid alphavirus in common dab Limanda limanda, with emphasis on virus culture and sequencing. Aquac Environ Interact. 2014;5: 89–98. doi:10.3354/aei00097en
dc.relation.hasversionNoguera PA, Bruno DW. Liver involvement in post-smolt Atlantic salmon, Salmo salar L., infected with infectious pancreatic necrosis virus (IPNV): A retrospective histopathological study. J Fish Dis. 2010;33: 819–832.en
dc.relation.hasversionCollet B, Urquhart K, Noguera P, Larsen KH, Lester K, Smail D, et al. A method to measure an indicator of viraemia in Atlantic salmon using a reporter cell line. J Virol Methods [Internet]. 2013 Aug [cited 2016 Apr 12];191(2):113–7. Available from: http://www.sciencedirect.com/science/article/pii/S0166093413001353en
dc.relation.hasversionNoguera PA, Grunow B, Klinger M, Lester K, Collet B, del-Pozo J. Atlantic salmon cardiac primary cultures: An in vitro model to study viral host pathogen interactions and pathogenesis. Vaughan L, editor. PLoS One. Elsevier; 2017;12: e0181058. doi:10.1371/journal.pone.0181058en
dc.relation.hasversionNoguera P, Collet B, Klinger M, Örün H, del Pozo J. Use of Salmon Cardiac Primary Cultures (SCPCs) of different genotypes for comparative kinetics of mx expression, viral load and ultrastructure pathology, after infection with Salmon Pancreas Disease Virus (SPDV). Fish Shellfish Immunol. 2018;72: 181–186. doi:10.1016/j.fsi.2017.10.059en
dc.subjectAtlantic salmonen
dc.subjectcardiotropic infectionsen
dc.subjectSPDVen
dc.subjectdisease modelen
dc.subjecthost pathogen interactionen
dc.subjectcardiac primary culturesen
dc.titleIn vivo and in vitro studies of Salmon Pancreas Disease Virus (SPDV) in Atlantic salmon (Salmo salar L.)en
dc.typeThesis or Dissertationen
dc.type.qualificationlevelDoctoralen
dc.type.qualificationnamePhD Doctor of Philosophyen
dc.rights.embargodate2019-11-30
dcterms.accessRightsRestricted Accessen


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