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  • 1
    ISSN: 1432-0568
    Keywords: Key words Monoclonal antibody ; Monocytes/macrophages ; Fish ; Ontogeny
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  A monoclonal antibody against carp macrophages (WCL15) has been utilised in flow cytometry, immuno-histochemistry and immuno-electron microscopy to assess the distribution of monocytes/macrophages in developing carp lymphoid tissues. In suspensions of living cells WCL15 reacted strongly with cytoplasm and plasmic membrane of macrophages. It also cross-reacted with a subpopulation of thrombocytes, but this reaction could be neglected by double immunostaining in combination with a thrombocyte-specific marker. In Bouin-fixed tissues the antibody distinctly recognised macrophages. Macrophages were found from day 2 post-fertilisation in head kidney and in the dorsal portion of the yolk sac epithelium. From 1 week onwards macrophages were found scattered in thymus and gut and during the second week in spleen. Macrophages increased in number in all lymphoid tissues until the 6–8th week post-fertilisation, but they decreased except in thymus, where they became localised mainly in the cortical-medullary boundary, and in white pulp areas of head kidney. The role of macrophages in allowing an early non-specific defence in young fish and in co-operating during the differentiation processes of T-cells and B-cells is discussed.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Journal of fish biology 49 (1996), S. 0 
    ISSN: 1095-8649
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Monoclonal antibodies (WCL6) specific for carp Cyprinus carpio thrombocytes were produced by immunizing mice with membrane lysates of IgM-negative peripheral blood leucocytes (PBL) and selected on the negative reaction with B cells. WCL6 was reactive with a membrane molecule of approximately 90 kDa and to a lesser extent with molecules of approximately 95 and 110 kDa. In general, between 30 and 40% of PBL were WCL6+ and appeared to be round to spindle-shaped cells. Immunohistochemical analysis of cryo-sections showed much higher numbers of WCL6+ cells in the spleen than in the pronephros, intestine and thymus. Flow cytometric analysis of cell suspensions isolated from these organs only revealed WCL6+ cells (4–10%) in the spleen. Electron microscopy of immunogold-stained WCL6+ PBL showed round but also some spindle-shaped cells with canalicular and granular structures, and a more irregular and electron-dense nucleus than found in lymphocytes. WCL6+ cells with electrondense pyknotic nuclei (without a clear nuclear envelope) were found also and their frequency increased with the length of the isolation and staining procedure used. In the spleen, several differentiation steps of WCL6+ cells were found and hence the spleen seems to be the thrombopoietic organ in carp. Thrombocytes from blood could be activated with collagen; the collagen-activated cells showed a higher side (90°) scatter by flow cytometric analysis and finally considerable cell death.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Journal of fish biology 35 (1989), S. 0 
    ISSN: 1095-8649
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Uptake and transport of soluble (ferritin) and particulate (Vibrio anguillarum) antigen from intestinal lumen to mucosal macrophages was studied using immunocytochemical and electron-microscopical techniques. Both antigens were taken up by epithelial cells of the second gut segment, reached the supranuclear vacuoles and were finally transported to large intraepithelial macrophages. In contrast with particulate antigen, antigenic determinants of ferritin were demonstrated in the basal part of the epithelium and appeared to be released at the mucosal site.After anal intubation many small macrophages penetrated the intestinal epithelium, took up antigen and after 24 h disappeared again from the intestinal mucosa. This feature was antigen independent and also occurred after anal PBS-intubation. The larger, less mobile macrophages stayed in the intestinal epithelium and finally exposed antigenic determinants (of both antigens) at their outer surface, suggesting an antigen-presenting function. Whether these large macrophages can induce a local or mucosal immune response and whether the smaller mobile macrophages are involved in a systemic response is discussed.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1095-8649
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Mucosal and systemic (serum) immune responses were studied after oral, anal or intramuscular (i.m.) immunization with particulate (Vibrio anguillarum) or soluble (ferritin) antigen. Antigen specific antibodies were found by ELISA in skin mucus after repeated oral or anal administration of bacteria, but not after immunization with ferritin. Daily feeding with bacteria did not give detectable antibodies in serum, while regular oral administration of ferritin resulted in an increase of specific antibodies during the first 3 weeks. From that time immunosuppression was observed, as the antibody titre decreased despite the continued ferritin feeding. Immunosuppression was also found after a second anal intubation or i.m. injection with ferritin, independent of the route of priming (i.m. or anal). On the contrary, a second anal intubation of bacteria resulted in a secondary serum response. These results combined with those reported in Parts I and II of the study indicate an important immunological role for the second gut segment. Because mucosal as well as serum responses can be obtained by anal immunization with bacteria, the significance for oral vaccination is discussed.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Journal of fish biology 35 (1989), S. 0 
    ISSN: 1095-8649
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Lymphocytes, plasma cells, granulocytes (three to four types), macrophages and monocyte-like cells were ultrastructurally distinguished in the intestinal mucosa of carp. Neutrophilic granulocytes and lymphoid cells were present in and under the epithelium throughout the gut. In contrast to macrophages which dominated in the epithelium of the second segment, basophilic and eosinophilic granulocytes (and their intermediates) were mainly found in the connective tissue of the first segment. Applying monoclonal antibodies against serum immunoglobulin (Ig) in an immunogold technique, only a minority of lymphoid cells appeared to be Ig-immunoreactive at their external membrane, suggesting the presence of many more T than B cells in the intestinal mucosa. Except for cells which resembled immature plasma cells, plasma cells did not show, or hardly showed, Ig at their surface. In contrast with the head kidney, plasma cells with an Ig-immunoreactive cytoplasm were scarce in the intestinal mucosa. As mucosa plasma cells were regularly found with the electron microscope, they possibly contain another class of Ig. Macrophages and monocyte-like cells were also found to be Ig-immunoreactive, suggesting the presence of immune complexes at their external membrane. The immunological significance of B- and T-like lymphocytes next to immune complex-binding and antigen-presenting macrophages in the second gut segment is discussed.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1433-2981
    Keywords: Antibacterial drugs ; Eel ; Fish culture ; Immunomodulation ; Leucocytes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In two experiments with European eel (Anguilla anguilla L., 1758) the effects of flumequine (FQ), oxytetracycline (OTC) and furazolidone (FZ) on peripheral blood leucocytes were tested using measurements of differential white blood cell counts in blood smears, flow cytometry and respiratory burst activity of adherent cells. Results revealed that FQ and OTC affected (different) leucocyte populations, whereas no effect of FZ was detected in these experiments. These effects should be taken into account when drugs are registered for official approval in veterinary (fish) medicine.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Using the semi-thin/ultra-thin technique six different immunoreactive endocrine cell types are ultrastructurally identified in 0.5% glutaraldehyde fixed gut of B. conchonius. In addition two of them (gastrin-and PP-immunoreactive cells) are also characterized with the immunogold method, showing that the immunoreactivity is only restricted to the secretory granules. Size distribution histograms and the average diameters of 30% (d30) of the largest granules are given, showing a gradual increase in granule size from unspecific immunoreactive cells, (d30=110 nm) via gastrin-(119 nm), VIP-like-(127 nm), met-enkephalin-(143 nm) and PP-(174 nm) to glucagon-immunoreactive cells (178 nm). The presence of PP-and glucagon-immunoreactivity in the same cells and the consequence for their granule size is discussed. In the distal part of the gut endocrine cells are found showing no immunoreactivity with the antisera used; their granules (d30=144 nm) were, although not significantly, larger then those of VIP-like-immunoreactive cells, also found in that part of the gut. It is supposed that they represent substance P-immunoreactive cells. Unfortanately, secretory granules of several cell types showed about 20% more shrinkage in 0.5% glutaraldehyde fixed tissue, than in osmicated tissue.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunocytochemical double staining techniques were used to study PP- and glucagon-like-immunoreactivity in pancreatic endocrine cells of mouse. An antiserum against FMRFamide appeared to react with all PP-immunoreactive endocrine cells. With fluorescence microscopy most PP/FMRFamide-immunoreactive cells also showed glucagon-immunoreactivity, but cells containing only PP-or glucagon-like substances were found as well. The proportion of cells containing PP-, glucagon, and both immunoreactivities varied strongly from islet to islet in all parts of the pancreas. Using an electron microscopical immunogold double staining procedure on Lowicryl-embedded pancreas, PP/FMRFamide-and glucagon-immunoreactivity appeared to be present in the majority of endocrine A cells; both immunoreactivities were randomly distributed within the granules of these cells. Cells containing only PP/FMRFamide-or glucagon-immunoreactivity were also found. Glucagon-and a faint FMRFamide-immunoreactivity was also observed in osmicated epon-embedded tissue. Independent of their immunoreactivity all positive cells showed the same round electron dense secretory granules.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1432-0878
    Keywords: Pancreatic endocrine cells ; Enteroendocrine cells ; Ultrastructure ; Cyprinidae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The pancreatic endocrine cells of Barbus conchonius are concentrated in a large (principal) islet, located near the gall bladder, and in a number of smaller islets. Five types of endocrine cells can be distinguished in these pancreatic islets: B cells, A1 (or D cells), 2 types of A2 cells (A2r cells with round granules; and A2fl cells with flocculent granules) and a scarce 5th cell type. The hormones produced by B and A2fl cells are probably insulin and glucagon respectively. The A2r cell contains granules with the same diameter as the granules of the enteroendocrine type III cell of the gut. Both cell types may resemble the enteroglucagon-producing EG cell of mammals. The function of the A1 cells, which are frequently found without secretory granules, and of the 5th cell type, will be discussed. The pancreatic islets of B. conchonius are strongly innervated, which suggests the presence of a direct nervous control system. Some intermediate or mixed cells containing exocrine and endocrine A2r granules are found contiguous with the principal islet. The origin of pancreatic endocrine cells is also the subject of discussion.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1432-0878
    Keywords: Development ; Enterocytes ; Fish ; Mitosis ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The processes of proliferation, cell division and differentiation of intestinal epithelial cells have been studied during development of the fish, Barbus conchonius. On the 3rd day, nearly all cells of the presumptive gut proliferate. Once the intestinal epithelium begins to differentiate, a decreasing percentage of proliferative cells can be found. On the 7th day, when intestinal folds start to develop, the proliferative cells become restricted to the future basal parts of the folds. Ultrastructural examination of 3H-thymidine-labeled cells and mitotic cells of 6-day-old larvae shows that functional enterocytes are proliferative. The same feature is suggested for older fish. Proliferating undifferentiated “dark” cells, characterized by many free ribosomes and a few organelles, are also present in the intestinal epithelium of larval fish; they are considered to be stem cells, mainly for goblet cells. Proliferating goblet cells and enteroendocrine cells were not observed. The latter cell type is scarce and has a long turnover time. A common feature of all these dividing cells is the presence of isolated spherical to cylindrical lamellar structures which may have lost contact with the cell membrane during prophase; they probably regain this contact by fusion with the cell membrane at the end of mitosis.
    Type of Medium: Electronic Resource
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