Infectious Salmon Anemia Virus RNA in Fish Cell Cultures and in Tissue Sections of Atlantic Salmon Experimentally Infected with Infectious Salmon Anemia Virus
Current understanding of the etiopathogenesis of infectious salmon anemia (ISA) virus (ISAV) infection in fish comes mostly from virus detection in homogenized tissues taken from ISA-suspected mortalities. This study combined in situ hybridization (ISH) and histology to demonstrate viral RNA transcr...
Published in: | Journal of Veterinary Diagnostic Investigation |
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Online Access: | http://dx.doi.org/10.1177/104063870301500502 https://journals.sagepub.com/doi/pdf/10.1177/104063870301500502 |
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crsagepubl:10.1177/104063870301500502 2024-10-06T13:47:18+00:00 Infectious Salmon Anemia Virus RNA in Fish Cell Cultures and in Tissue Sections of Atlantic Salmon Experimentally Infected with Infectious Salmon Anemia Virus Moneke, Emeka E. Kibenge, Molly J. T. Groman, David Johnson, Gerald R. Ikede, Basil O. Kibenge, Frederick S. B. 2003 http://dx.doi.org/10.1177/104063870301500502 https://journals.sagepub.com/doi/pdf/10.1177/104063870301500502 en eng SAGE Publications https://journals.sagepub.com/page/policies/text-and-data-mining-license Journal of Veterinary Diagnostic Investigation volume 15, issue 5, page 407-417 ISSN 1040-6387 1943-4936 journal-article 2003 crsagepubl https://doi.org/10.1177/104063870301500502 2024-09-10T04:25:35Z Current understanding of the etiopathogenesis of infectious salmon anemia (ISA) virus (ISAV) infection in fish comes mostly from virus detection in homogenized tissues taken from ISA-suspected mortalities. This study combined in situ hybridization (ISH) and histology to demonstrate viral RNA transcripts in different fish cell lines infected with ISAV and in tissues collected during the clinical phase of ISAV infection in Atlantic salmon. For this, a riboprobe to mRNA transcripts of ISAV RNA segment 8 was shown to detect viral mRNA in ISAV-infected TO, CHSE-214, and SHK-1 cell cultures. Specific hybridization was initially detected exclusively in the nuclei of infected cells, which is consistent with the nuclear transcription of orthomyxoviruses. For use of the riboprobe on fish tissues fixed in paraformaldehyde or formalin, the conditions used to permeabilize tissues before ISH (Proteinase K or Tween 20) were first optimized. Tissues were collected 15—20 days after challenge from 7 fresh mortalities of Atlantic salmon parr (∼20 g) showing severe gross and microscopic lesions, consistent with ISAV infection. Reverse transcription-polymerase chain reaction on tissue pools confirmed the presence of ISAV in each of the 7 fish. Of the tissues examined in each fish, the heart and liver consistently showed the strongest hybridization signal and, therefore, the most in situ virus, which was located in the endothelium of small blood vessels and in macrophage-like cells. Article in Journal/Newspaper Atlantic salmon SAGE Publications Journal of Veterinary Diagnostic Investigation 15 5 407 417 |
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English |
description |
Current understanding of the etiopathogenesis of infectious salmon anemia (ISA) virus (ISAV) infection in fish comes mostly from virus detection in homogenized tissues taken from ISA-suspected mortalities. This study combined in situ hybridization (ISH) and histology to demonstrate viral RNA transcripts in different fish cell lines infected with ISAV and in tissues collected during the clinical phase of ISAV infection in Atlantic salmon. For this, a riboprobe to mRNA transcripts of ISAV RNA segment 8 was shown to detect viral mRNA in ISAV-infected TO, CHSE-214, and SHK-1 cell cultures. Specific hybridization was initially detected exclusively in the nuclei of infected cells, which is consistent with the nuclear transcription of orthomyxoviruses. For use of the riboprobe on fish tissues fixed in paraformaldehyde or formalin, the conditions used to permeabilize tissues before ISH (Proteinase K or Tween 20) were first optimized. Tissues were collected 15—20 days after challenge from 7 fresh mortalities of Atlantic salmon parr (∼20 g) showing severe gross and microscopic lesions, consistent with ISAV infection. Reverse transcription-polymerase chain reaction on tissue pools confirmed the presence of ISAV in each of the 7 fish. Of the tissues examined in each fish, the heart and liver consistently showed the strongest hybridization signal and, therefore, the most in situ virus, which was located in the endothelium of small blood vessels and in macrophage-like cells. |
format |
Article in Journal/Newspaper |
author |
Moneke, Emeka E. Kibenge, Molly J. T. Groman, David Johnson, Gerald R. Ikede, Basil O. Kibenge, Frederick S. B. |
spellingShingle |
Moneke, Emeka E. Kibenge, Molly J. T. Groman, David Johnson, Gerald R. Ikede, Basil O. Kibenge, Frederick S. B. Infectious Salmon Anemia Virus RNA in Fish Cell Cultures and in Tissue Sections of Atlantic Salmon Experimentally Infected with Infectious Salmon Anemia Virus |
author_facet |
Moneke, Emeka E. Kibenge, Molly J. T. Groman, David Johnson, Gerald R. Ikede, Basil O. Kibenge, Frederick S. B. |
author_sort |
Moneke, Emeka E. |
title |
Infectious Salmon Anemia Virus RNA in Fish Cell Cultures and in Tissue Sections of Atlantic Salmon Experimentally Infected with Infectious Salmon Anemia Virus |
title_short |
Infectious Salmon Anemia Virus RNA in Fish Cell Cultures and in Tissue Sections of Atlantic Salmon Experimentally Infected with Infectious Salmon Anemia Virus |
title_full |
Infectious Salmon Anemia Virus RNA in Fish Cell Cultures and in Tissue Sections of Atlantic Salmon Experimentally Infected with Infectious Salmon Anemia Virus |
title_fullStr |
Infectious Salmon Anemia Virus RNA in Fish Cell Cultures and in Tissue Sections of Atlantic Salmon Experimentally Infected with Infectious Salmon Anemia Virus |
title_full_unstemmed |
Infectious Salmon Anemia Virus RNA in Fish Cell Cultures and in Tissue Sections of Atlantic Salmon Experimentally Infected with Infectious Salmon Anemia Virus |
title_sort |
infectious salmon anemia virus rna in fish cell cultures and in tissue sections of atlantic salmon experimentally infected with infectious salmon anemia virus |
publisher |
SAGE Publications |
publishDate |
2003 |
url |
http://dx.doi.org/10.1177/104063870301500502 https://journals.sagepub.com/doi/pdf/10.1177/104063870301500502 |
genre |
Atlantic salmon |
genre_facet |
Atlantic salmon |
op_source |
Journal of Veterinary Diagnostic Investigation volume 15, issue 5, page 407-417 ISSN 1040-6387 1943-4936 |
op_rights |
https://journals.sagepub.com/page/policies/text-and-data-mining-license |
op_doi |
https://doi.org/10.1177/104063870301500502 |
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Journal of Veterinary Diagnostic Investigation |
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15 |
container_issue |
5 |
container_start_page |
407 |
op_container_end_page |
417 |
_version_ |
1812175531685183488 |