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

Bertrand Collet, Katy Urquhart, Milena Monte, Catherine Collins, Sandro Garcia Perez, Chris J Secombes, Malcolm Hall

Research output: Contribution to journalArticle

13 Citations (Scopus)
5 Downloads (Pure)

Abstract

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 IP 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 IFN could be detected. These results and the advantages of this approach are discussed.
Original languageEnglish
Article numbere0137767
JournalPloS ONE
Volume10
Issue number9
DOIs
Publication statusPublished - 23 Sep 2015

Fingerprint

Isavirus
Infectious salmon anemia virus
Salmo salar
Immunologic Monitoring
Viruses
Fish
Fishes
Animals
Blood
immune response
Monitoring
monitoring
Infection
fish
Gene expression
infection
stress response
blood
animals
Immunology

Cite this

Individual monitoring of immune response in Atlantic salmon Salmo salar following experimental infection with Infectious Salmon Anaemia Virus (ISAV). / Collet, Bertrand; Urquhart, Katy; Monte, Milena; Collins, Catherine; Garcia Perez, Sandro; Secombes, Chris J; Hall, Malcolm.

In: PloS ONE, Vol. 10, No. 9, e0137767, 23.09.2015.

Research output: Contribution to journalArticle

Collet, Bertrand ; Urquhart, Katy ; Monte, Milena ; Collins, Catherine ; Garcia Perez, Sandro ; Secombes, Chris J ; Hall, Malcolm. / Individual monitoring of immune response in Atlantic salmon Salmo salar following experimental infection with Infectious Salmon Anaemia Virus (ISAV). In: PloS ONE. 2015 ; Vol. 10, No. 9.
@article{9b6e1929f47d4c9fbce676687f0bd5a8,
title = "Individual monitoring of immune response in Atlantic salmon Salmo salar following experimental infection with Infectious Salmon Anaemia Virus (ISAV)",
abstract = "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 IP 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 IFN could be detected. These results and the advantages of this approach are discussed.",
author = "Bertrand Collet and Katy Urquhart and Milena Monte and Catherine Collins and {Garcia Perez}, Sandro and Secombes, {Chris J} and Malcolm Hall",
note = "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.",
year = "2015",
month = "9",
day = "23",
doi = "10.1371/journal.pone.0137767",
language = "English",
volume = "10",
journal = "PloS ONE",
issn = "1932-6203",
publisher = "PUBLIC LIBRARY SCIENCE",
number = "9",

}

TY - JOUR

T1 - Individual monitoring of immune response in Atlantic salmon Salmo salar following experimental infection with Infectious Salmon Anaemia Virus (ISAV)

AU - Collet, Bertrand

AU - Urquhart, Katy

AU - Monte, Milena

AU - Collins, Catherine

AU - Garcia Perez, Sandro

AU - Secombes, Chris J

AU - Hall, Malcolm

N1 - 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.

PY - 2015/9/23

Y1 - 2015/9/23

N2 - 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 IP 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 IFN could be detected. These results and the advantages of this approach are discussed.

AB - 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 IP 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 IFN could be detected. These results and the advantages of this approach are discussed.

U2 - 10.1371/journal.pone.0137767

DO - 10.1371/journal.pone.0137767

M3 - Article

VL - 10

JO - PloS ONE

JF - PloS ONE

SN - 1932-6203

IS - 9

M1 - e0137767

ER -