This work was funded by the National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs, grant G1100675). The authors are grateful to the Marine Scotland aquarium staff (Ben Williamson, Louise Feehan and Mark Paterson) for help in the experimental work. Hendrix-Genetix (formerly Landcatch Natural Selection) is also acknowledged for providing PIT tagged fish for the purpose of this project.Peer reviewedPublisher PD

Collet, Bertrand

Collins, Catherine

Garcia Perez, Sandro

Hall, Malcolm

Monte, Milena

Secombes, Chris J

Urquhart, Katy

PLoS ONE

PubMed

This work was funded by the National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs, grant G1100675). The authors are grateful to the Marine Scotland aquarium staff (Ben Williamson, Louise Feehan and Mark Paterson) for help in the experimental work. Hendrix-Genetix (formerly Landcatch Natural Selection) is also acknowledged for providing PIT tagged fish for the purpose of this project.Peer reviewe

Aberdeen University Research

Individual monitoring of immune response in Atlantic salmon Salmo salar following experimental infection with Infectious Salmon Anaemia Virus (ISAV)

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

Bertrand Collet

Katy Urquhart

Milena Monte

Catherine Collins

Sandro Garcia Perez

Chris J Secombes

Malcolm Hall

Directory of Open Access Journals

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

peer reviewedMonitoring 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 gammaIP 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 gamma IFN could be detected. These results and the advantages of this approach are discussed

Mira Monte, Milena

Secombes, Chris J.

Open Repository and Bibliography - Liège

Chris J. Secombes

Crossref

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

<div><p>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 (<i>Salmo salar</i>) were i.p infected with Infectious Salmon Anaemia Virus (ISAV) or culture medium and placed in 1 m<sup>3</sup> 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.</p></div

Bertrand Collet (485023)

Katy Urquhart (799009)

Milena Monte (799010)

Catherine Collins (186960)

Sandro Garcia Perez (799011)

Chris J. Secombes (421713)

Malcolm Hall (799012)

The Francis Crick Institute

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

http://orbi.ulg.ac.be/bitstream/2268/205288/1/journal.pone.0137767.pdf

Individual monitoring of immune response in Atlantic salmon Salmo salar following experimental infection with Infectious Salmon Anaemia Virus (ISAV)

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Aberdeen University Research

Directory of Open Access Journals

Open Repository and Bibliography - Liège

Crossref

The Francis Crick Institute