Detection of the myxosporean parasite Parvicapsula pseudobranchicola in Atlantic salmon (Salmo salar L.) using in situ hybridization (ISH)
Abstract Background Parvicapsula pseudobranchicola is a marine myxosporean parasite infecting farmed Atlantic salmon (Salmo salar). A major site for the parasite is the pseudobranch, which may be destroyed in heavily infected fish. Parvicapsulosis may be associated with significant mortality, althou...
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ftdoajarticles:oai:doaj.org/article:a8a624d2f5494ffdbc2f2fd98714ba39 2023-07-02T03:31:41+02:00 Detection of the myxosporean parasite Parvicapsula pseudobranchicola in Atlantic salmon (Salmo salar L.) using in situ hybridization (ISH) Turhan Markussen Celia Agusti Egil Karlsbakk Are Nylund Øyvind Brevik Haakon Hansen 2015-02-01T00:00:00Z https://doi.org/10.1186/s13071-015-0718-4 https://doaj.org/article/a8a624d2f5494ffdbc2f2fd98714ba39 EN eng BMC https://doi.org/10.1186/s13071-015-0718-4 https://doaj.org/toc/1756-3305 doi:10.1186/s13071-015-0718-4 1756-3305 https://doaj.org/article/a8a624d2f5494ffdbc2f2fd98714ba39 Parasites & Vectors, Vol 8, Iss 1, Pp 1-6 (2015) Parvicapsula pseudobranchicola Myxozoa Atlantic salmon Parasites in situ hybridization ISH Infectious and parasitic diseases RC109-216 article 2015 ftdoajarticles https://doi.org/10.1186/s13071-015-0718-4 2023-06-11T00:37:44Z Abstract Background Parvicapsula pseudobranchicola is a marine myxosporean parasite infecting farmed Atlantic salmon (Salmo salar). A major site for the parasite is the pseudobranch, which may be destroyed in heavily infected fish. Parvicapsulosis may be associated with significant mortality, although the main effect of infections seems to be runting. In situ hybridization (ISH) is, in the absence of specific antibodies, the preferred method for the detection of cell- and tissue tropisms of myxozoans in the early phases of infection of the host, and provides information about the possible association between the pathogen and pathology. A positive diagnosis of parvicapsulosis is based on histopathology and PCR. The aim of the present work was to develop a specific, sensitive and robust ISH assay for the detection of P. pseudobranchicola in tissues. Methods The ISH method was designed to specifically target P. pseudobranchicola 18S rDNA/rRNA using a locked nucleic acid (LNA) modified oligonucleotide probe. The method was tested on paraffin embedded P. pseudobranchicola infected pseudobranchs. The infections were confirmed by light microscopy revealing the presence of typical P. pseudobranchicola trophozoites and spores, and the presence of parasite was confirmed with real-time RT-PCR. Results Specific regions stained by ISH overlapped well with the parasitized and degenerated regions in neighbouring HE stained sections. No staining was observed in pseudobranchs of Atlantic salmon which had been held in P. pseudobranchicola-free water. Conclusions We report here the development of a sensitive ISH assay for the detection of P. pseudobranchicola in paraffin embedded tissue. The technique will be valuable in the study of host entry, early proliferation, pre-spore development, pathology and tissue tropism in Atlantic salmon. Article in Journal/Newspaper Atlantic salmon Salmo salar Directory of Open Access Journals: DOAJ Articles Parasites & Vectors 8 1 105 |
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English |
topic |
Parvicapsula pseudobranchicola Myxozoa Atlantic salmon Parasites in situ hybridization ISH Infectious and parasitic diseases RC109-216 |
spellingShingle |
Parvicapsula pseudobranchicola Myxozoa Atlantic salmon Parasites in situ hybridization ISH Infectious and parasitic diseases RC109-216 Turhan Markussen Celia Agusti Egil Karlsbakk Are Nylund Øyvind Brevik Haakon Hansen Detection of the myxosporean parasite Parvicapsula pseudobranchicola in Atlantic salmon (Salmo salar L.) using in situ hybridization (ISH) |
topic_facet |
Parvicapsula pseudobranchicola Myxozoa Atlantic salmon Parasites in situ hybridization ISH Infectious and parasitic diseases RC109-216 |
description |
Abstract Background Parvicapsula pseudobranchicola is a marine myxosporean parasite infecting farmed Atlantic salmon (Salmo salar). A major site for the parasite is the pseudobranch, which may be destroyed in heavily infected fish. Parvicapsulosis may be associated with significant mortality, although the main effect of infections seems to be runting. In situ hybridization (ISH) is, in the absence of specific antibodies, the preferred method for the detection of cell- and tissue tropisms of myxozoans in the early phases of infection of the host, and provides information about the possible association between the pathogen and pathology. A positive diagnosis of parvicapsulosis is based on histopathology and PCR. The aim of the present work was to develop a specific, sensitive and robust ISH assay for the detection of P. pseudobranchicola in tissues. Methods The ISH method was designed to specifically target P. pseudobranchicola 18S rDNA/rRNA using a locked nucleic acid (LNA) modified oligonucleotide probe. The method was tested on paraffin embedded P. pseudobranchicola infected pseudobranchs. The infections were confirmed by light microscopy revealing the presence of typical P. pseudobranchicola trophozoites and spores, and the presence of parasite was confirmed with real-time RT-PCR. Results Specific regions stained by ISH overlapped well with the parasitized and degenerated regions in neighbouring HE stained sections. No staining was observed in pseudobranchs of Atlantic salmon which had been held in P. pseudobranchicola-free water. Conclusions We report here the development of a sensitive ISH assay for the detection of P. pseudobranchicola in paraffin embedded tissue. The technique will be valuable in the study of host entry, early proliferation, pre-spore development, pathology and tissue tropism in Atlantic salmon. |
format |
Article in Journal/Newspaper |
author |
Turhan Markussen Celia Agusti Egil Karlsbakk Are Nylund Øyvind Brevik Haakon Hansen |
author_facet |
Turhan Markussen Celia Agusti Egil Karlsbakk Are Nylund Øyvind Brevik Haakon Hansen |
author_sort |
Turhan Markussen |
title |
Detection of the myxosporean parasite Parvicapsula pseudobranchicola in Atlantic salmon (Salmo salar L.) using in situ hybridization (ISH) |
title_short |
Detection of the myxosporean parasite Parvicapsula pseudobranchicola in Atlantic salmon (Salmo salar L.) using in situ hybridization (ISH) |
title_full |
Detection of the myxosporean parasite Parvicapsula pseudobranchicola in Atlantic salmon (Salmo salar L.) using in situ hybridization (ISH) |
title_fullStr |
Detection of the myxosporean parasite Parvicapsula pseudobranchicola in Atlantic salmon (Salmo salar L.) using in situ hybridization (ISH) |
title_full_unstemmed |
Detection of the myxosporean parasite Parvicapsula pseudobranchicola in Atlantic salmon (Salmo salar L.) using in situ hybridization (ISH) |
title_sort |
detection of the myxosporean parasite parvicapsula pseudobranchicola in atlantic salmon (salmo salar l.) using in situ hybridization (ish) |
publisher |
BMC |
publishDate |
2015 |
url |
https://doi.org/10.1186/s13071-015-0718-4 https://doaj.org/article/a8a624d2f5494ffdbc2f2fd98714ba39 |
genre |
Atlantic salmon Salmo salar |
genre_facet |
Atlantic salmon Salmo salar |
op_source |
Parasites & Vectors, Vol 8, Iss 1, Pp 1-6 (2015) |
op_relation |
https://doi.org/10.1186/s13071-015-0718-4 https://doaj.org/toc/1756-3305 doi:10.1186/s13071-015-0718-4 1756-3305 https://doaj.org/article/a8a624d2f5494ffdbc2f2fd98714ba39 |
op_doi |
https://doi.org/10.1186/s13071-015-0718-4 |
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Parasites & Vectors |
container_volume |
8 |
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1 |
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105 |
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