Characterization of early phases of cardiomyopathy syndrome pathogenesis in Atlantic salmon (Salmo salar L.) through various diagnostic methods

Figures

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  FIGURE 1. Histopathology scores of the three cardiac compartments atrium, ventricular spongiosum and ventricular compactum for all sampling time points, in accordance with Table 2. Average values plus standard deviation of the mean (SD) are shown (n = 5 at 0‐day post‐challenge (dpc), n = 15 at 10 and 20 dpc and n = 13 at 52 dpc)

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  FIGURE 2. Normal atrium and ventricular spongiosum found at 0 dpc (exemplified by fish [F] 1 and 2, respectively), and typical CMS lesions (arrows) in atrium and ventricular spongiosum found at 52 dpc in F9 and 15, respectively. HE‐staining and standard light microscopy at 200x magnification (bar = 250 μm)

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  FIGURE 3. Levels of PMCV‐specific RNA given as Cq values, for all sampling time points, in days post‐challenge (dpc) in three heart compartments, mid‐kidney and skin/muscle and blood components and surface mucus. (a) Tissue samples of cardiac atrium, ventricular spongiosum, ventricular compactum, mid‐kidney and skin/muscle. (b) Parallel samples of blood: whole blood, plasma and blood cell pellet. (c) Mucus samples at three different locations of the surface of the fish: Pectoral fins, lateral line and anus. Results are presented as box plot with whiskers representing minimum and maximum values in the individual data set. n = 4 at 0 dpc and n = 15 at other time points, excluding ventricular spongiosum at 52 dpc where n = 4. Note the inverted numbering of the Y‐axis

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  FIGURE 4. Examples from RNAscope in situ hybridization of cardiac atrium and ventricular spongiosum, represented by hearts of fish F1, F7, F2 and F1 at 0, 10, 20 and 52 dpc, respectively, and mid‐kidney represented by F3, F3, F2 and F2 at 0, 10, 20 and 52 dpc, respectively. Detection of PMCV‐specific RNA is visible as dark red staining (arrows in images of sparse staining). S = spongiosum, T = lumen of tubuli, M = melanin deposits and/or melanomacrophages in the interstitium of the kidney. All images 400× magnification, bar = 50 μm, standard light microscopy

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  FIGURE 5. Details of RNAscope in situ hybridization results. Detection of PMCV‐specific RNA is visible as dark red staining. (a) Staining in endothelial nuclei in a coronary vessel (arrows). L = lumen of coronary vessel. (b) Staining (arrow) in atrial myocardial cells. Note the lack of staining in the myocardial nucleus (MN). Both images, 400 x magnification, bar = 50 μm, standard light microscopy