Development of immunohistochemistry and enzyme-linked immunosorbent assays for the detection of circulating antibodies against Enteromyxum scophthalmi (Myxozoa) in turbot (Scophthalmus maximus L.)
Immunohistochemistry and enzyme-linked immunosorbent assays were developed for the detection of specific antibodies against the myxosporean parasite Enteromyxum scophthalmi in turbot (Scophthalmus maximus L.). Fish which had survived a previous epizootic were exposed to the parasite by cohabitation...
Published in: | Fish & Shellfish Immunology |
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Format: | Article in Journal/Newspaper |
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Elsevier
2004
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Online Access: | http://hdl.handle.net/10261/253756 https://doi.org/10.1016/j.fsi.2004.04.007 https://doi.org/10.13039/501100000780 https://doi.org/10.13039/501100006280 |
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ftcsic:oai:digital.csic.es:10261/253756 2024-02-11T10:08:26+01:00 Development of immunohistochemistry and enzyme-linked immunosorbent assays for the detection of circulating antibodies against Enteromyxum scophthalmi (Myxozoa) in turbot (Scophthalmus maximus L.) Sitjà-Bobadilla, Ariadna Redondo, Mª José Macías, M. A. Ferreiro, Isabel Riaza, Ana Álvarez-Pellitero, Mª del Pilar European Commission Stolt Sea Farm Ministerio de Ciencia y Tecnología (España) 2004-10 http://hdl.handle.net/10261/253756 https://doi.org/10.1016/j.fsi.2004.04.007 https://doi.org/10.13039/501100000780 https://doi.org/10.13039/501100006280 unknown Elsevier http://dx.doi.org/10.1016/j.fsi.2004.04.007 Sí doi:10.1016/j.fsi.2004.04.007 issn: 1050-4648 Fish and Shellfish Immunology 17(4): 335-345 (2004) http://hdl.handle.net/10261/253756 http://dx.doi.org/10.13039/501100000780 http://dx.doi.org/10.13039/501100006280 none Aquaculture ELISA Enteromyxosis Humoral immune response Immunohistochemistry Myxosporea Turbot artículo http://purl.org/coar/resource_type/c_6501 2004 ftcsic https://doi.org/10.1016/j.fsi.2004.04.00710.13039/50110000078010.13039/501100006280 2024-01-16T11:15:14Z Immunohistochemistry and enzyme-linked immunosorbent assays were developed for the detection of specific antibodies against the myxosporean parasite Enteromyxum scophthalmi in turbot (Scophthalmus maximus L.). Fish which had survived a previous epizootic were exposed to the parasite by cohabitation with infected animals, and 83 days later the plasma was tested for the presence of antibodies. Plasma of non-exposed fish was used as negative control. Immunohistochemistry (IHC) using rabbit anti-turbot IgM antibody was first used to detect these antibodies, and to study to which parasite structures they were directed against. Also, an antibody-ELISA using whole cell lysates of the parasite as antigen, and a monoclonal antibody anti-turbot IgM, was developed. All the exposed fish were found to have specific antibodies against the parasite, and none of them developed signs of disease or died during the experiment. Primary cells were the main parasite stage immunolabelled, and the staining was distinctly located on the cytoplasm and the cytoplasmic membrane. IHC was more sensitive than ELISA, as the endpoint was two to four fold higher with the former technique. Although there was great individual variation, the antibody titres found can be considered high, reaching up to 1:32,000 with ELISA and 1:64,000 with IHC. The results suggest that turbot showing acquired immunity against E. scophthalmi, could develop resistance against new infections. This study was funded by the FEDER research grant 1FD97-0679-C02-01 (European Union, Spanish Government and Stolt Sea Farm, S.A.) and research project AGL2001-2241-C01-01 (Spanish Ministry of Science and Technology). Additional funding was provided by Stolt Sea Farm, S.A. through a research contract. We are grateful to J. Monfort for the histological processing, and to Dr. M.I. Quiroga from the Veterinary Faculty of Lugo (University of Santiago) for providing some paraffin blocks. Article in Journal/Newspaper Scophthalmus maximus Turbot Digital.CSIC (Spanish National Research Council) Fish & Shellfish Immunology 17 4 335 345 |
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Open Polar |
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Digital.CSIC (Spanish National Research Council) |
op_collection_id |
ftcsic |
language |
unknown |
topic |
Aquaculture ELISA Enteromyxosis Humoral immune response Immunohistochemistry Myxosporea Turbot |
spellingShingle |
Aquaculture ELISA Enteromyxosis Humoral immune response Immunohistochemistry Myxosporea Turbot Sitjà-Bobadilla, Ariadna Redondo, Mª José Macías, M. A. Ferreiro, Isabel Riaza, Ana Álvarez-Pellitero, Mª del Pilar Development of immunohistochemistry and enzyme-linked immunosorbent assays for the detection of circulating antibodies against Enteromyxum scophthalmi (Myxozoa) in turbot (Scophthalmus maximus L.) |
topic_facet |
Aquaculture ELISA Enteromyxosis Humoral immune response Immunohistochemistry Myxosporea Turbot |
description |
Immunohistochemistry and enzyme-linked immunosorbent assays were developed for the detection of specific antibodies against the myxosporean parasite Enteromyxum scophthalmi in turbot (Scophthalmus maximus L.). Fish which had survived a previous epizootic were exposed to the parasite by cohabitation with infected animals, and 83 days later the plasma was tested for the presence of antibodies. Plasma of non-exposed fish was used as negative control. Immunohistochemistry (IHC) using rabbit anti-turbot IgM antibody was first used to detect these antibodies, and to study to which parasite structures they were directed against. Also, an antibody-ELISA using whole cell lysates of the parasite as antigen, and a monoclonal antibody anti-turbot IgM, was developed. All the exposed fish were found to have specific antibodies against the parasite, and none of them developed signs of disease or died during the experiment. Primary cells were the main parasite stage immunolabelled, and the staining was distinctly located on the cytoplasm and the cytoplasmic membrane. IHC was more sensitive than ELISA, as the endpoint was two to four fold higher with the former technique. Although there was great individual variation, the antibody titres found can be considered high, reaching up to 1:32,000 with ELISA and 1:64,000 with IHC. The results suggest that turbot showing acquired immunity against E. scophthalmi, could develop resistance against new infections. This study was funded by the FEDER research grant 1FD97-0679-C02-01 (European Union, Spanish Government and Stolt Sea Farm, S.A.) and research project AGL2001-2241-C01-01 (Spanish Ministry of Science and Technology). Additional funding was provided by Stolt Sea Farm, S.A. through a research contract. We are grateful to J. Monfort for the histological processing, and to Dr. M.I. Quiroga from the Veterinary Faculty of Lugo (University of Santiago) for providing some paraffin blocks. |
author2 |
European Commission Stolt Sea Farm Ministerio de Ciencia y Tecnología (España) |
format |
Article in Journal/Newspaper |
author |
Sitjà-Bobadilla, Ariadna Redondo, Mª José Macías, M. A. Ferreiro, Isabel Riaza, Ana Álvarez-Pellitero, Mª del Pilar |
author_facet |
Sitjà-Bobadilla, Ariadna Redondo, Mª José Macías, M. A. Ferreiro, Isabel Riaza, Ana Álvarez-Pellitero, Mª del Pilar |
author_sort |
Sitjà-Bobadilla, Ariadna |
title |
Development of immunohistochemistry and enzyme-linked immunosorbent assays for the detection of circulating antibodies against Enteromyxum scophthalmi (Myxozoa) in turbot (Scophthalmus maximus L.) |
title_short |
Development of immunohistochemistry and enzyme-linked immunosorbent assays for the detection of circulating antibodies against Enteromyxum scophthalmi (Myxozoa) in turbot (Scophthalmus maximus L.) |
title_full |
Development of immunohistochemistry and enzyme-linked immunosorbent assays for the detection of circulating antibodies against Enteromyxum scophthalmi (Myxozoa) in turbot (Scophthalmus maximus L.) |
title_fullStr |
Development of immunohistochemistry and enzyme-linked immunosorbent assays for the detection of circulating antibodies against Enteromyxum scophthalmi (Myxozoa) in turbot (Scophthalmus maximus L.) |
title_full_unstemmed |
Development of immunohistochemistry and enzyme-linked immunosorbent assays for the detection of circulating antibodies against Enteromyxum scophthalmi (Myxozoa) in turbot (Scophthalmus maximus L.) |
title_sort |
development of immunohistochemistry and enzyme-linked immunosorbent assays for the detection of circulating antibodies against enteromyxum scophthalmi (myxozoa) in turbot (scophthalmus maximus l.) |
publisher |
Elsevier |
publishDate |
2004 |
url |
http://hdl.handle.net/10261/253756 https://doi.org/10.1016/j.fsi.2004.04.007 https://doi.org/10.13039/501100000780 https://doi.org/10.13039/501100006280 |
genre |
Scophthalmus maximus Turbot |
genre_facet |
Scophthalmus maximus Turbot |
op_relation |
http://dx.doi.org/10.1016/j.fsi.2004.04.007 Sí doi:10.1016/j.fsi.2004.04.007 issn: 1050-4648 Fish and Shellfish Immunology 17(4): 335-345 (2004) http://hdl.handle.net/10261/253756 http://dx.doi.org/10.13039/501100000780 http://dx.doi.org/10.13039/501100006280 |
op_rights |
none |
op_doi |
https://doi.org/10.1016/j.fsi.2004.04.00710.13039/50110000078010.13039/501100006280 |
container_title |
Fish & Shellfish Immunology |
container_volume |
17 |
container_issue |
4 |
container_start_page |
335 |
op_container_end_page |
345 |
_version_ |
1790607770798522368 |