Vaccine-associated granulomatous inflammation and melanin accumulation in Atlantic salmon, Salmo salar L., white muscle.

The purpose of this study was to investigate the nature of variably sized pigmented foci encountered in fillets of farmed Atlantic salmon, Salmo salar L. The material was sampled on the fillet production line and on salmon farms from fish with an average size of 3 kg from various producers. The fish had been routinely vaccinated by injection. Gross pathology, histology, immunohistochemistry using antisera against major histocompatibility complex (MHC) class II beta chain and transmission electron microscopy (TEM) were used to characterize the changes. Macroscopically, melanized foci were seen penetrating from the peritoneum deep into the abdominal wall, sometimes right through to the skin, and also embedded in the caudal musculature. Histological investigation revealed muscle degeneration and necrosis, fibrosis and granulomatous inflammation containing varying numbers of melano-macrophages. Vacuoles, either empty or containing heterogeneous material, were frequently seen. The presence of abundant MHC class II+ cells indicated an active inflammatory condition. TEM showed large extracellular vacuoles and leucocytes containing homogeneous material of lipid-like appearance. The results showed that the melanized foci in Atlantic salmon fillet resulted from an inflammatory condition probably induced by vaccination. The described condition is not known in wild salmon and in farmed salmon where injection vaccination is not applied.

Characterization of macrophages and occurrence of T cells in intestinal lesions of subclinical paratuberculosis in goats.

The granulomatous lesions of subclinical paratuberculosis of goats were examined with emphasis on phenotypic characteristics of macrophages and the presence of different subpopulations of T cells. The macrophages in the granulomatous lesions were morphologically homogeneous in histological sections but showed varying expression of the macrophage marker CD68 (a glycoprotein found mainly in late endosomal and lysosomal membranes) and varying acid phosphatase activity. The lesional macrophages showed decreased expression of complement receptor 3 and major histocompatibility complex proteins, which are markers associated with phagocytosis and antigen-presentation, respectively. The granulomas showed low proliferation activity as measured by the proliferation-associated protein Ki-67, indicating that most cells were recruited to the lesions. Few apoptotic cells were demonstrated by the TUNEL technique, suggesting a low cell turnover in the lesions. CD4(+) T cells constituted the main T-cell population among the CD68(+) macrophages in the granulomatous lesions, and few CD8(+) T cells and gamma delta T cells were observed within the lesions, suggesting the limited ability of these cells to influence the granulomatous lesions in caprine subclinical paratuberculosis. Both WC1(+) and WC1(-) gamma delta T cells were present in the small intestinal wall, but the latter were the more numerous. No difference in the numbers of these cells was observed between the subclinically infected animals and control animals.

Production of rabbit antisera against recombinant MHC class II beta chain and identification of immunoreactive cells in Atlantic salmon (Salmo salar).

In the present work, rabbit antisera recognising the Atlantic salmon (Salmo salar) MHC class II beta chain polypeptide were produced and used in immunoblotting, immunohistochemistry and immunogold electron microscopy. A construct encoding the beta1 and beta2 domains fused to the E. coli protein thioredoxin was used to express the recombinant MHC class II beta chain. Immunoblotting revealed a band of approximately 30kDa in total protein samples from head kidney, spleen, gills, thymus and blood leukocytes, while being absent in muscle. The distribution of MHC class II positive cells was immunohistochemically demonstrated in Atlantic salmon epithelial and haemopoietic tissues. Ultrastructural demonstration of immunoreactive organelles in mid-kidney cells was performed by immunogold electron microscopy. The results indicate expression in lymphocytes, macrophages, epithelial cells and endritic-like cells. This is the first study to address morphological MHC class II expression in a fish species.