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A direct monoclonal antibody sandwich immunoassay for detection of Bonamia ostreae (Ascetospora) in hemolymph samples of the flat oyster Ostrea edulis (Mollusca : Bivalvia) ArchiMer
Cochennec, Nathalie; Hervio, Dominique; Panatier, B; Boulo, Viviane; Mialhe, Eric; Rogier, H; Grizel, Henri; Paolucci, F.
Currently recognition of molluscan diseases relies heavily upon light microscopic examination of stained histological sections of molluscan tissues Such histopathological methods are time consuming and relatively inefficient In the study of ongoing epizootics Alternatively, more effluent investigative tools are needed to accelerate disease recognition and quantification Such methods are needed to prevent control and eradicate molluscan diseases This paper reports the development of a direct sandwich enzyme-lmmunosorbent assay (ELISA) for the ascetosporan intrahemocytic parasite Bonamiao ostreae of the flat oyster Ostrea edulis. Bonamiasis is a commercially important epizootic disease of oysters A Bonamia-specific monoclonal antibody is employed in the...
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Ano: 1992 URL: http://archimer.ifremer.fr/doc/1992/publication-2765.pdf
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In vitro culture of mollusc hemocytes. Functional study of burst respiratory activity and analysis of interactions with protozoan and procaryotic pathogens ArchiMer
Boulo, Viviane; Hervio, Dominique; Morvan, Annie; Bachere, Evelyne; Mialhe, Eric.
The chief importance of hemocytes as immune effectors in molluscs has led to develop researches for in vitro culture. The chemiluminescence technique has been used to estimate the phagocytosis capacity of Crassostrea gigas hemocytes according to different culture media and times. Sea water was used as a basic medium. Additives, such as antibiotics or glucose and buffers, such as Tris or Hepes, were tested as weIl as artificial sea water. Hemocyte chemiluminescence activities were recorded at t=o in relation to the medium composition. The highest values were observed when hemocytes were kept in sea water without any modification. Whichever medium, chemiluminescence activity was substantially decreased when hemocytes were previously kept in vitro for a few...
Tipo: Text Palavras-chave: Crassotrea gigas; Oyster; Bonamia ostreae; Pathogens; Molluscs; Hemocytes.
Ano: 1991 URL: http://archimer.ifremer.fr/doc/1991/acte-4115.PDF
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Localisation and characterization of acid phosphate activity in Bonamia ostreae (Ascetospora), an intrahemocytic protozoan parasite of the flat oyster Ostrea edulis (Bivalvia) ArchiMer
Hervio, Dominique; Chagot, Dominique; Godin, Pierre; Grizel, Henri; Mialhe, Eric.
Acid phosphatase activity was cytochemically detected at the ultrastructural level in the protozoan Bonamia ostreae, an intrahemocytic parasite of the flat oyster Ostrea edulis. Lead nitrate was used as capture agent for phosphate liberated during the hydrolysis of the subtrate (cytidine 5' monophosphate). Lead phosphate precipiation, indicative of enzyme activity, was located in membrane-bound organelles corresponding to the cytoplasmic inclusions known as "dense bodies". Acid phosphatase activity level and sodium L-tartrate inhibition were measured spectrophotometrically from a purified parasite suspension. The possible involvement of this enzyme in intracellular survival of B. ostrea is discussed.
Tipo: Text Palavras-chave: Acid phosphate activity; Ostrea edulis; Bonamia ostreae.
Ano: 1991 URL: http://archimer.ifremer.fr/doc/1991/publication-2755.pdf
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Localization and characterization of acid phosphatase activity in Bonamia ostreae (Ascetospora), an intrahemocytic protozoan parasite of the fIat oyster Ostrea edulis (Bivalvia) ArchiMer
Hervio, Dominique; Chagot, D; Godin, P; Grizel, Henri; Mialhe, Eric.
Acid phosphatase activity was cytochemically detected at the ultrastructural level in the protozoan Bonamia ostreae, an intrahemocytic parasite of the flat oyster Ostrea edulis. Lead nitrate was used as capture agent for phosphate liberated during the hydrolysis of the substrate (cytidine 5' monophosphate). Lead phosphate precipitation, indicative of enzyme activity, was located in membrane-bound organelles corresponding to the cytoplasmic inclusions known as 'dense bodies'. Acid phosphatase activity level and sodium L-tartrate inhibition were measured spectrophotometrically from a purified parasite suspension. The possible involvement of this enzyme in intracellular survival of B. ostreae is discussed.
Tipo: Text
Ano: 1991 URL: http://archimer.ifremer.fr/doc/00182/29311/28273.pdf
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Luminol-dependent chemiluminescence by hemocytes of two marine bivalves, Ostrea edulis and Crassostrea gigas ArchiMer
Bachere, Evelyne; Hervio, Dominique; Mialhe, Eric.
Microbicidal phagocytic function and oxidative metabolism of the hemocytes of 2 oyster species, Ostrea edulis and Crassostrea gigas, were investigated using a luminol-enhanced chemiluminescence (technique. First, experimental parameters adapted to marine bivalve hemocytes were established on hemolymph pools of C. gigas in order to obtain qualitatively and quantitatively homogeneous samples and so to perform statistically comparable assays. The use of Modified Alsever Solution (allowed hemocytes to be kept non-aggregated and in a non-stimulated state until starting the assays. A number of aliquots of 2 X 10(5) hemocytes with an MAS final concentration of 3.5 % showed high chemiluminescent response after stimulation by zymosan particles A particle:hemocyte...
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Ano: 1991 URL: http://archimer.ifremer.fr/doc/1991/publication-2761.pdf
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Monoclonal antibodies against Bonamia ostreae (Protozoa: Ascetospera), an intrahaemocytic parasite of flat oyster Ostrea edulis (Mollusca : Bivlavia) ArchiMer
Rogier, Hervé; Hervio, Dominique; Boulo, Viviane; Clavies, Christine; Hervaud, Eliane; Bachere, Evelyne; Mialhe, Eric; Grizel, Henri; Pau, Bernard; Paolucci, Francis.
The protozoan Bonamia ostreae (Ascetospora), a paasite of the flat oyster Ostrea edulis, was purified by differential and isopycnic centrifugations. Mice of the strain Balb/c were immunized with purified parasites and hybridomas were prepared by fusion of immunized mouse splenocytes with the mouse myeloma cell line P3-X63-Ag8-653. From the fusion, 12 clones were isolated and saved. The resulting antibodies were characterized with a solid phase radioimmunoassay (RIA). Antibodies from 7 clones reacted only with B. ostreae and not with normal host tissue. Although some of the antibiodies identified. One of these antibodies (20B2-1B12) has an apparent association constant of ca 3 X 10(8) M-1 and bound to a number of different sites on the parasite. These...
Tipo: Text Palavras-chave: Monoclonal antibodies; Oyster; Ostrea edulis; Bonamia ostreae.
Ano: 1991 URL: http://archimer.ifremer.fr/doc/1991/publication-2756.pdf
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Strategy of researches on marine bivalve defense mechanisms ArchiMer
Bachere, Evelyne; Mialhe, Eric; Noel, Thierry; Boulo, Viviane; Hervio, Dominique; Le Gall, Ghyslaine; Morvan, Annie.
Because of the impact of infectious diseases in marine mollusc aquaculture, research has been initially developed in pathology in term of epidemiology and diagnosis. The establishment of basic techniques for in vitro and in vivo studies of host/pathogen interactions and the progresses in the field of cellular and molecular biology have permitted to undertake researches in immunology. Parallel to fundamental interest in understanding defense mecanisms of molluscs against pathogens, applications may be expected for resistant strains obtained by genetic engineering.
Tipo: Text Palavras-chave: Defense mechanisms; Hosts; Immunology; Pathogens; Crassostrea gigas; Mytilus edulis.
Ano: 1991 URL: http://archimer.ifremer.fr/doc/00038/14884/12207.pdf
Registros recuperados: 7
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