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Farmed Atlantic salmon (Salmo salar) are prone to various conditions affecting the quality of the fillet. A well-known but so far poorly understood condition is the focal red changes in muscle, often referred to as haemorrhages. Such changes are characterized by muscle necrosis, haemorrhages and acute inflammation. They can progress into focal melanised changes, a chronic inflammatory condition with melanin-producing leukocytes. The initial cause of intramuscular haemorrhages is unknown. In this study, we aimed to reveal some of their key immunological features. Samples of red focal changes were investigated by immunohistochemistry (IHC), in situ hybridization (ISH) and RT-qPCR for various immune markers. The results were compared with samples of melanised changes and control muscle, subjected to the same analyses. In all red changes, infiltrates with mononuclear cells were detected, consisting mostly of MHC class I/II + cells, but also of CD3 + and CD8 + cells. ISH studies on IgM showed few to moderate amounts of B-cells in red focal changes. Trends in the RT-qPCR showed upregulation of genes related to innate immunity in the red changes, whereas genes related to adaptive immunity were upregulated in the melanised changes. An important result was the significant downregulation of the anti-inflammatory cytokine IL10 in all red changes. Our findings indicate that we can rule out an auto invasive nature of the changes. The downregulation of IL10 at an early phase is a trait for the condition., Competing Interests: Declaration of Competing Interest The authors declare that they have no competing interests., (Copyright © 2020 The Author(s). Published by Elsevier B.V. All rights reserved.)

ABSTRACT Melanisation can occur in the musculature of fish. A well‐known form is the melanised focal changes, or ‘black spots’, in the fillet of farmed Atlantic salmon (Salmo salar). The aetiology of black spots has not been fully determined, though recent research has emphasised the role of fat necrosis in their development. The initial stages of the changes are observed as focal haemorrhages or ‘red spots’, and these can progress into melanised focal changes (MFCs). The focal haemorrhages are acute changes characterised by necrotic myocytes and adipocytes and diffuse haemorrhage in the tissue. These changes evolve into a chronic inflammation dominated by fibrosis, encapsulated lipid droplets or pseudocysts, presence of epithelioid cells, granulomas of varying character, giant cells and melano‐macrophages, whose presence accounts for the discolouration. The inflammation ranges from mild to severe, and the severity of the lesion has been associated with localised piscine orthoreovirus 1 (PRV 1) replication in macrophages and melano‐macrophages within granulomas. The possibility of a genetic impact on the condition has not been supported by available data. The lipid composition and the antioxidative properties of the feed have been shown to affect the development of the changes. Physiological and environmental factors are also believed to influence the prevalence and severity of the condition. Here, we review the current state of knowledge concerning melanisation in fish skeletal musculature, with a special emphasis on the MFCs in Atlantic salmon. [ABSTRACT FROM AUTHOR]

Melanized focal changes in skeletal muscle of farmed Atlantic salmon (Salmo salar) are a major quality problem. The aetiology is unknown, but infection with Piscine orthoreovirus (PRV) has been associated with the condition. Here, we addressed the pathogenesis of red and melanized focal changes and their association with PRV. First, a population of farmed fish (PRV‐negative prior to sea transfer) was sequentially investigated throughout the seawater period. The fish were autopsied and tested for PRV infection. Muscular changes were described by macroscopy and histology, and a classification system was established. Second, in an experimental infection trial, PRV was injected intramuscularly to induce changes. The farmed fish was gradually infected with PRV. Red focal changes occurred throughout the observation period with a low prevalence regardless of PRV status. Melanized changes were highly diverse and their prevalence increased during the trial. Changes of low macroscopic grade and histological category were more prevalent in PRV‐negative fish. Diffuse granulomatous melanized changes only occurred after PRV infection. No muscular changes were observed in the experimentally challenged fish. Our studies do not indicate that PRV infection causes red focal changes, but seems important in the development of granulomatous melanized changes. [ABSTRACT FROM AUTHOR]

Abstract: The heart is considered the powerhouse of the cardiovascular system. Heart and skeletal muscle inflammation (HSMI), cardiomyopathy syndrome (CMS) and pancreas disease (PD) are cardiac diseases of marine farmed Atlantic salmon (Salmo salar) which commonly affect the heart in addition to the skeletal muscle, liver and pancreas. The main findings of these diseases are necrosis and inflammatory cells infiltrates affecting different regions of the heart. In order to better characterize the cardiac pathology, study of the inflammatory cell characteristics and cell cycle protein expression was undertaken by immunohistochemistry. Immunohistochemistry was performed on paraffin embedded hearts from confirmed diseased cases applying specific antibodies. The inflammatory cells were predominantly CD3+ T lymphocytes. The PD diseased hearts exhibited moderate hypoxia inducible factor-1α (HIF1α) immuno-reaction that suggested tissue hypoxia while recombinant tumor necrosis factor-α (rTNFα) antibody identified putative macrophages and eosinophilic granulocytes (EGCs) in addition to endocardial cells around lesions. There were strong to low levels of major histocompatibility complex (MHC) class II immunostaining in the diseased hearts associated with macrophage-like and lymphocyte-like cells. The diseased hearts expressed strong to low levels of apoptotic cells identified by caspase 3 and terminal deoxynucleotidyl transferase nick-end labeling (TUNEL) staining. The strong signals for proliferative cell nuclear antigen (PCNA) and TUNEL, and moderate levels of caspase 3 immuno-reactivity suggested a high cell turnover where DNA damage/repair might be occurring in the diseased hearts. Interestingly, the apparently similar cardiac diseases exhibited differences in the immunopathological responses in Atlantic salmon. [Copyright &y& Elsevier]

• IgD mRNA was the most expressed Ig in hind-gut suffering feed-induced inflammation. IgM RNA expression was also abundant. • IgT-expressing cells were scarce. • IgD-positive cells could be involved in mucosal hypersensitivity reactions caused by food allergens. • Dislocated epithelial cells were found in association with inflammation, as has been reported in previous studies. [ABSTRACT FROM AUTHOR]

Abstract: Development of diagnostic and prophylactic methodologies is dependent on knowledge of the host’s defence system and reaction to different vaccine adjuvants. Here we present a sequential morphological study of peritonitis and inflammatory cell processing of incomplete Freund’s adjuvant (IFA) in intraperitoneally injected Atlantic cod. The peritoneal tissue responses were characterised using necropsy, histology and electron microscopy. An extensive inflammatory response as characterised by leukocyte morphology and contents of enzymes, presence of apoptotic cells and IFN-γ-expressing cells was observed. Three days post injection, IFA droplets were surrounded by different types of inflammatory cells and two different patterns could be discerned. The first was characterised by flattened and concentrically arranged interdigitating cells connected by desmosomes and with macrophage-like cells (MLCs) predominant in the periphery. The second type possessed four stratified layers with an inner layer containing many apoptotic MLCs; a second layer containing flattened and shrunken cells and outer layers comprising moderately flattened cells and an outermost layer of mononuclear cells expressing IFN-γ. Oil was detected both inside and outside MLCs. The two types of processes, of which the second was clearly stratified, were similar to those observed in other teleosts, indicating a variety of reaction modes or alternatively sequential process development. The numerous dead MLCs contributed to inflammation. [Copyright &y& Elsevier]

Animal models are invaluable tools in cancer research. In this context, salmon is a promising candidate. Intestinal adenocarcinoma with metastases may be induced as a consequence of a plant-based diet triggering the inflammation – dysplasia- carcinogenesis pathway. Here, we investigate the stroma and the presence and nature of immune cells in such tumors by staining for mast cells, immunohistochemistry for T cells and antigen-presenting cells and in situ hybridization for B cells. In intestinal tumors, substantial amounts of T cells were detected in the stroma, whilst MHC class II+ cells were mainly among the cancerous cells. Ig+ cells were observed primarily in the tumor periphery. Mast cells showed a strong association with stroma. In metastases, scarce amounts of T cells were detected, whilst MHC I and II-reactivity varied, some tumors being completely negative. Ig+ cells were scattered around the metastatic tissue in no particular pattern, but were occasionally observed within clusters of tumor cells. Small numbers of mast cells were detected in the stroma. To the best of our knowledge, this is the first report addressing immune cells in fish tumors. The teleost tumor microenvironment seems comparable to that of mammals, making fish interesting model animals in oncoimmunology research. [ABSTRACT FROM AUTHOR]