Stimulation of nonspecific defense mechanisms and the specific immune response in fish
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Geology topics
Publications and source records attributed to D. P. Anderson.
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No abstract available at this time
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To determine the earliest time after in vivo immunisation that the spleen could be excised and held in vitro to detect an immune response, lake trout ( Salvelinus namaycush ) were exposed to DNP-Ficoll or Yersinia ruckeri O antigen administered by intraperitoneal injection or by bath. The spleens were excised from the fish at selected times after immunisation, placed in vitro and held in tissue culture media at 14°C until 10 days after the in vivo immunisation. They then were analysed for an immune response by counting the numbers of plaqueforming cells (PFC). PFC were first detected in samples taken 3 days after injection with either antigen. With bath immunisation, however, PFC were first seen 6 days post-immunisation when using the Y. ruckeri O antigen and no PFC above background levels appeared in fish bathed in the DNP-Ficoll solution. On day 10 after immunisation, the spleens taken directly from immunised fish always produced more PFC than when the spleens were taken through in vitro culture. A unique feature of this method, is that an immune response to an antigen may be detected by excising the spleen and holding it in in vitro culture without necessarily holding the fish for long periods of time after antigen injection or bath. This technique also adds more information on how rapidly the bacterins or antigens are processed by fish to initiate an immune response.
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Review of: Systemic Pathology of Fish: A Text and Atlas of Comparative Tissue Responses in Diseases of Teleosts. By Hugh W. Ferguson. Iowa State University Press, Ames. 1989. 263 pages. $54.95.
The spleen and head kidney of striped bass Morone saxatilis were examined by electron microscopy, including examination of ultrastructural localization of peroxidase. The spleen is composed of a loosely organized reticulum that supports the red and white pulp. The principal hematopoietic and mature blood cells observed were erythrocytes, erythroblasts, lymphocytes, macrophages, thrombocytes, and aggregates of pigmented macrophages. In addition, limited numbers of neutrophils, neutrophilic myelocytes (immature neutrophils), plasma cells, eosinophils and monocytes were identified. Reticular cells and fibers appeared to provide a structural framework for both cells and blood sinuses in the spleen. Like the spleen, the head kidney contained erythrocytes, macrophages, monocytes, lymphocytes, and other less frequently observed cell types. A distinct difference in the cytologic composition of the two organs, however, was in the greater number of neutrophilic myelocytes found in the head kidney, where these cells were often arranged in groups. Positive peroxidase reactions were observed for neutrophils, neutrophilic myelocytes, macrophage phagolysosomes, and pigmented macrophages in both the spleen and head kidney.
A cell line from rainbow trout ( Oncorhynchus mykiss ) with phagocytic-like characteristics was established. First we found that it was necessary to inject the fish with antibiotics (penicillinstreptomycin) in order to clear tissue of bacterial contamination. Two days later, the spleen was excised and minced with scissors in MEM10. The large number of erythrocytes in the suspension were removed by low centrifugation, then the tissue fragments were incubated on tissue culture flasks at 18°C for 30 days with complete changes of media every other day. After 30 days in culture, primary explants from spleen were composed of three types of cells. The majority were large mononucleate, plecmorphic cells with thinly spread cytoplasm. These cells were trypsinized and subcultured. Another cell types was simple, round cells, somewhat similar to mature lymphocytes, while the third type appeared to be similar to fibroblastic cells. After 9 months with the culture in 17 to 20 passages, the predominate cell type was showing similarities to phagocytic cell. These cells had a marked reticular cytoplasm and grew individually without parallel orientations. By enzyme-specific stains, the cells were observed to have abundant acid phosphatase and β-glucuronidase activities of the cytoplasm. They were negative for peroxidase. The cells were also found to have highly adherent and phagocytic properties when incubated with latex particles. The morphological, enzymatic, and functional findings suggested the possibility that these cells in culture have some properties similar to the cells which are composed of reticulo-endothelial system.