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      Histopathological investigation of complex gill disease in sea farmed Atlantic salmon

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          Abstract

          Various agents including Ca. Piscichlamydia salmonis, Ca. Branchiomonas cysticola, Desmozoon lepeophtherii, Paramoeba perurans and salmon gill poxvirus may be associated with complex gill disease in Atlantic salmon. Co-infections involving two or more of these agents are common and histopathological interpretation of lesions is therefore challenging. In this study, we developed a semi-quantitative scoring system for examination of histopathological gill lesions in sea-farmed Atlantic salmon suffering from gill disease. Following qPCR analysis of gills sampled for Ca. P. salmonis, Ca. B. cysticola, D. lepeophtherii and P. perurans from 22 geographically spread outbreaks, five cases representing different infectious loads and combinations of agents were chosen for histopathological scoring. Twenty-eight histological features were evaluated and potential associations between individual pathological changes and the occurrence of individual agents studied. The inter-observer agreement in interpretation of histological parameters between the three pathologists involved, was calculated to validate robustness of the scoring scheme. Seventeen histological parameters met the criteria for inter-observer agreement analysis and were included in the calculation. The three most frequent findings were identification of subepithelial leukocytes, epithelial cell hyperplasia and mucus cell hyperplasia. While few findings could be specifically related to particular agents, necrosis in hyperplastic lesions, pustules and necrosis of subepithelial cells appeared to be associated with the presence of Ca. B. cysticola. Further, lesion profiles clearly support the previously identified association between P. perurans and pathological changes associated with AGD. Very few pathological changes were observed in the single case in which Ca. P. salmonis was the dominating agent. Some lesions were only very rarely observed e.g. chloride cell necrosis, epithelial cell apoptosis, lamellar deposition of melanin and haemophagocytosis. The scoring scheme developed and applied was robust and sensitive. A less extensive scheme for routine diagnostic use is proposed.

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          Interrater agreement and interrater reliability: key concepts, approaches, and applications.

          Evaluations of interrater agreement and interrater reliability can be applied to a number of different contexts and are frequently encountered in social and administrative pharmacy research. The objectives of this study were to highlight key differences between interrater agreement and interrater reliability; describe the key concepts and approaches to evaluating interrater agreement and interrater reliability; and provide examples of their applications to research in the field of social and administrative pharmacy. This is a descriptive review of interrater agreement and interrater reliability indices. It outlines the practical applications and interpretation of these indices in social and administrative pharmacy research. Interrater agreement indices assess the extent to which the responses of 2 or more independent raters are concordant. Interrater reliability indices assess the extent to which raters consistently distinguish between different responses. A number of indices exist, and some common examples include Kappa, the Kendall coefficient of concordance, Bland-Altman plots, and the intraclass correlation coefficient. Guidance on the selection of an appropriate index is provided. In conclusion, selection of an appropriate index to evaluate interrater agreement or interrater reliability is dependent on a number of factors including the context in which the study is being undertaken, the type of variable under consideration, and the number of raters making assessments. Copyright © 2013 Elsevier Inc. All rights reserved.
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            A review of infectious gill disease in marine salmonid fish.

            Infectious gill diseases of marine salmonid fish present a significant challenge in salmon-farming regions. Infectious syndromes or disease conditions affecting marine-farmed salmonids include amoebic gill disease (AGD), proliferative gill inflammation (PGI) and tenacibaculosis. Pathogens involved include parasites, such as Neoparamoeba perurans, bacteria, such as Piscichlamydia salmonis and Tenacibaculum maritimum, and viruses, such as the Atlantic salmon paramyxovirus (ASPV). The present level of understanding of these is reviewed with regard to risk factors, potential impacting factors, methods of best practice to mitigate infectious gill disease, as well as knowledge gaps and avenues for future research. © 2011 Blackwell Publishing Ltd.
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              Neoparamoeba perurans n. sp., an agent of amoebic gill disease of Atlantic salmon (Salmo salar).

              Amoebic gill disease (AGD) is a potentially fatal disease of some marine fish. Two amphizoic amoebae Neoparamoeba pemaquidensis and Neoparamoeba branchiphila have been cultured from AGD-affected fish, yet it is not known if one or both are aetiological agents. Here, we PCR amplified the 18S rRNA gene of non-cultured, gill-derived (NCGD) amoebae from AGD-affected Atlantic salmon (Salmo salar) using N. pemaquidensis and N. branchiphila-specific oligonucleotides. Variability in PCR amplification led to comparisons of 18S rRNA and 28S rRNA gene sequences from NCGD and clonal cultured, gill-derived (CCGD) N. pemaquidensis and N. branchiphila. Phylogenetic analyses inferred from either 18S or 28S rRNA gene sequences unambiguously segregated a lineage consisting of NCGD amoebae from other members of the genus Neoparamoeba. Species-specific oligonucleotide probes that hybridise 18S rRNA were designed, validated and used to probe gill tissue from AGD-affected Atlantic salmon. The NCGD amoebae-specific probe bound AGD-associated amoebae while neither N. pemaquidensis nor N. branchiphila were associated with AGD-lesions. Together, these data indicate that NCGD amoebae are a new species, designated Neoparamoeba perurans n.sp. and this is the predominant aetiological agent of AGD of Atlantic salmon cultured in Tasmania, Australia.
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                Author and article information

                Contributors
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: VisualizationRole: Writing – original draft
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: InvestigationRole: Writing – original draft
                Role: ConceptualizationRole: Formal analysisRole: InvestigationRole: Writing – original draft
                Role: ConceptualizationRole: Formal analysisRole: InvestigationRole: Writing – original draft
                Role: Data curationRole: Formal analysisRole: Writing – original draft
                Role: ConceptualizationRole: Funding acquisitionRole: MethodologyRole: SupervisionRole: Writing – original draft
                Role: ConceptualizationRole: Formal analysisRole: Project administrationRole: ResourcesRole: Writing – original draft
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                3 October 2019
                2019
                : 14
                : 10
                : e0222926
                Affiliations
                [1 ] Norwegian Veterinary Institute, Oslo, Norway
                [2 ] Norwegian Veterinary Institute, Bergen, Norway
                Friedrich-Loeffler-Institut Bundesforschungsinstitut fur Tiergesundheit, GERMANY
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Author information
                http://orcid.org/0000-0001-5098-7905
                Article
                PONE-D-19-14344
                10.1371/journal.pone.0222926
                6776330
                31581255
                0b3c99f4-569f-4973-b065-61604711835f
                © 2019 Gjessing et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 21 May 2019
                : 10 September 2019
                Page count
                Figures: 6, Tables: 3, Pages: 18
                Funding
                Funded by: Norwegian Seafood Research Fund
                Award ID: 900800
                Funded by: Norwegian research council
                Award ID: 233858
                This work was supported by: MCG, Norwegian Seafood Research Fund project 900800: https://www.fhf.no/; Norwegian research council 233858: https://www.forskningsradet.no/. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Anatomy
                Animal Anatomy
                Aquatic Respiratory Anatomy
                Gills
                Medicine and Health Sciences
                Anatomy
                Animal Anatomy
                Aquatic Respiratory Anatomy
                Gills
                Biology and Life Sciences
                Zoology
                Animal Anatomy
                Aquatic Respiratory Anatomy
                Gills
                Biology and Life Sciences
                Anatomy
                Respiratory System
                Gills
                Medicine and Health Sciences
                Anatomy
                Respiratory System
                Gills
                Biology and Life Sciences
                Cell Biology
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                Epithelial Cells
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                Anatomy
                Biological Tissue
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                Medicine and Health Sciences
                Anatomy
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                Epithelium
                Epithelial Cells
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                Diagnostic Medicine
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                Necrosis
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