A method to measure an indicator of viraemia in Atlantic salmon using a reporter cell line

Collet, Bertrand; Urquhart, Katy; Noguera, Patricia A.; Larsen, Katrine H.; Lester, Katherine; Smail, D. A.; Bruno, D. W.

doi:10.1016/j.jviromet.2013.04.009

Cited by 12 publications

(9 citation statements)

References 34 publications

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“…It is therefore pertinent to investigate gene expression levels directly in the blood cells, to which the non-lethal model is restricted. In addition, a large number of immune parameters can be measured in small volume of plasma such as antibody levels, neutralisation activity of viraemia [ 35 ].…”

Section: Discussionmentioning

confidence: 99%

Individual Monitoring of Immune Response in Atlantic Salmon Salmo salar following Experimental Infection with Infectious Salmon Anaemia Virus (ISAV)

et al. 2015

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Monitoring the immune response in fish over the progression of a disease is traditionally carried out by experimental infection whereby animals are killed at regular intervals and samples taken. We describe here a novel approach to infectiology for salmonid fish where blood samples are collected repeatedly in a small group of PIT-tagged animals. This approach contributes to the reduction of animals used in research and to improved data quality. Two groups of 12 PIT-tagged Atlantic salmon (Salmo salar) were i.p infected with Infectious Salmon Anaemia Virus (ISAV) or culture medium and placed in 1 m3 tanks. Blood samples were collected at 0, 4, 8, 12, 16, 21 and 25 days post infection. The viral load, immune and stress response were determined in individual fish by real-time quantitative PCR (QPCR) on the blood cells, as well as the haematocrit used as an indicator of haemolysis, a clinical consequence of ISAV infection. “In-tank” anaesthesia was used in order to reduce the stress related to chase and netting prior to sampling. The data were analysed using a statistical approach which is novel with respect to its use in fish immunology. The repeated blood collection procedure did not induce stress response as measured by HSP70 and HSP90 gene expression in the un-infected animals. A strong increase in viraemia as well as a significant induction of Mx and γIP gene expression were observed in the infected group. Interleukin 10 was found induced at the later stage of the infection whereas no induction of CD8 or γ IFN could be detected. These results and the advantages of this approach are discussed.

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Section: Discussionmentioning

confidence: 99%

Individual Monitoring of Immune Response in Atlantic Salmon Salmo salar following Experimental Infection with Infectious Salmon Anaemia Virus (ISAV)

et al. 2015

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“…Mortality due these conditions varies, but all of them can also cause severe growth rate reduction and downgrading at harvest, with consequential impact on commercial value. These infections exhibit differences in cardiac immunopathological responses [25,26] and some important insights into host pathogen interactions have been gained through in vitro studies on non-cardiac cell lines [12,27,28]. However, understanding the basis of cardiac diseases in fish, as with other hosts, has been hampered by the lack of appropriate models that mimic the complexity of the organ [29].…”

Section: Introductionmentioning

confidence: 99%

Atlantic salmon cardiac primary cultures: An in vitro model to study viral host pathogen interactions and pathogenesis

et al. 2017

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Development of Salmon Cardiac Primary Cultures (SCPCs) from Atlantic salmon pre-hatch embryos and their application as in vitro model for cardiotropic viral infection research are described. Producing SCPCs requires plating of trypsin dissociated embryos with subsequent targeted harvest from 24h up to 3 weeks, of relevant tissues after visual identification. SCPCs are then transferred individually to chambered wells for culture in isolation, with incubation at 15–22°. SCPCs production efficiency was not influenced by embryo’s origin (0.75/ farmed or wild embryo), but mildly influenced by embryonic developmental stage (0.3 decline between 380 and 445 accumulated thermal units), and strongly influenced by time of harvest post-plating (0.6 decline if harvested after 72 hours). Beating rate was not significantly influenced by temperature (15–22°) or age (2–4 weeks), but was significantly lower on SCPCs originated from farmed embryos with a disease resistant genotype (F = 5.3, p<0.05). Two distinct morphologies suggestive of an ex vivo embryonic heart and a de novo formation were observed sub-grossly, histologically, ultra-structurally and with confocal microscopy. Both types contained cells consistent with cardiomyocytes, endothelium, and fibroblasts. Ageing of SCPCs in culture was observed with increased auto fluorescence in live imaging, and as myelin figures and cellular degeneration ultra-structurally. The SCPCs model was challenged with cardiotropic viruses and both the viral load and the mx gene expression were measurable along time by qPCR. In summary, SCPCs represent a step forward in salmon cardiac disease research as an in vitro model that partially incorporates the functional complexity of the fish heart.

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“…Gilthead seabream Mx promoters (pMx1, pMx2 and pMx3) were activated by poly I:C in suitably transfected RTG-2 cells (González-Mariscal et al, 2014), which are permissive to the infection by several fish viruses (Isshiki et al, 2001;Song et al, 2005;Ariel et al, 2009;Collet et al, 2013). Thus, RTG-2 cells are a validated experimental system for the functional characterization of virus-host interactions.…”

Section: Discussionmentioning

confidence: 99%

Gilthead seabream (Sparus aurata) Mx gene promoters respond differentially to IPNV and VHSV infections in RTG-2 cells

González-Mariscal

Fernández-Trujillo

Alonso

et al. 2016

Veterinary Immunology and Immunopathology

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A method to measure an indicator of viraemia in Atlantic salmon using a reporter cell line

Cited by 12 publications

References 34 publications

Individual Monitoring of Immune Response in Atlantic Salmon Salmo salar following Experimental Infection with Infectious Salmon Anaemia Virus (ISAV)

Individual Monitoring of Immune Response in Atlantic Salmon Salmo salar following Experimental Infection with Infectious Salmon Anaemia Virus (ISAV)

Atlantic salmon cardiac primary cultures: An in vitro model to study viral host pathogen interactions and pathogenesis

Gilthead seabream (Sparus aurata) Mx gene promoters respond differentially to IPNV and VHSV infections in RTG-2 cells

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