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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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Adaptation of the sea-bass (Dicentrarchus Labrax) to fresh water: Role of aquaporins and Na+/K+-ATPases ArchiMer
Giffard Mena, I; Lorin Nebel, C; Charmantier, G; Castille, R; Boulo, Viviane.
Sea-bass (Dicentrarchus labrax) grow under different salinity regimes, from the open sea to lagoons and even rivers, but some mortality has been recorded in juvenile stages when exposed to low salinity water. Changes in water permeability of different osmoregulatory tissues could be the cause of reduction in blood osmotic pressure and death in some fish in fresh water (FW). In order to explore this condition, we have studied the changes of aquaporins (AQP1 and AQP3), alpha 1 and alpha 4 Na+/K(+-)ATPase transcript levels in the digestive tract, kidney and gills after a long-term exposure of juvenile sea-bass to sea water (SW) and FW fish able to survive in SW and FW are called SW-adapted fish (SWS), FW successfully-adapted fish (FWS) respectively, while...
Tipo: Text Palavras-chave: Salinity adaptation; Juvenile sea bass; Differential mortality; Alpha 4 Na+/K+ ATPase; Alpha l Na+/K+ ATPase; AQP3; AQP1.
Ano: 2008 URL: http://archimer.ifremer.fr/doc/2008/publication-4656.pdf
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An infection of Ruditapes decussatus (Bivalvia) by Rickettsia ArchiMer
Mialhe, Eric; Chagot, Dominique; Boulo, Viviane; Comps, Michel; Ruano, Francisco; Grizel, Henri.
The authors report the presence of a Rickettsia in Ruditapes decussatus from natural populations of the Algarve (Portugal). Light microscopic shows infections of varying importance in gill tissues. Some enclose small procaryotic colonies (cp) scattered in all lamellae; in others, the colonies are scarce and hypertrophic (100 mu m). These different pictures suggest an amplification process. Digestive diverticulae may also be infected, but to a lesser extent. The ultrastructural features of this intravacuolar (mvd: densified vacuolar membrane) procaryote (P) and its multiplication by scission without a complex developmental cycle allows its classification in the order Rickettsiales.
Tipo: Text Palavras-chave: Ruditapes philippinarum; Pathogens; Histopathology; Pathology.
Ano: 1987 URL: http://archimer.ifremer.fr/doc/1987/acte-3107.pdf
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Bivale mollusc cell culture ArchiMer
Mialhe, Eric; Boulo, Viviane; Grizel, Henri.
The historical background of bivalve cell culture and its potential uses in molluscan pathology are reviewed. Then, primary cultures for the study of some host-pathogen interactions at the cellular level are considered. Next, alternative methods to classical cell cultures are examined that may lead to the production of bivalve celllines.
Tipo: Text Palavras-chave: Cell culture; Pathology; Molluscs.
Ano: 1988 URL: http://archimer.ifremer.fr/doc/1988/publication-3122.pdf
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Bonamiasis : a model study of diseases in marine molluscs ArchiMer
Grizel, Henri; Mialhe, Eric; Chagot, Dominique; Boulo, Viviane; Bachere, Evelyne.
Research on bonamiasis, an epizootic disease of the edible oyster Ostrea edu/is caused by the protozoan Bonamia ostreae, is discussed in relation to oyster fanning, research technology, epizootiology, and management. Morphological and infectious characteristics of the parasite are described. Recent progress in isolation and purification of the parasite have permitted investigations into host defense mechanisms, parasite infectivity, and the development of a mol1usc-pathogen mode!.
Tipo: Text Palavras-chave: Epizootic disease; Pathology; Protozoan; Bonamia ostreae; Oysters.
Ano: 1988 URL: http://archimer.ifremer.fr/doc/1988/publication-3120.pdf
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Cellular and molecular hemocyte responses of the Pacific oyster, Crassostrea gigas, following bacterial infection with Vibrio aestuarianus strain 01/32 ArchiMer
Labreuche, Yannick; Lambert, Christophe; Soudant, Philippe; Boulo, Viviane; Huvet, Arnaud; Nicolas, Jean-louis.
The strategies used by bacterial pathogens to circumvent host defense mechanisms remain largely undefined in bivalve molluscs. In this study, we investigated experimentally the interactions between the Pacific oyster (Crassostrea gigas) immune system and Vibrio aestuarianus strain 01/32, a pathogenic bacterium originally isolated from moribund oysters. First, an antibiotic-resistant V. aestuarianus strain was used to demonstrate that only a limited number of bacterial cells was detected in the host circulatory system, suggesting that the bacteria may localize in some organs. Second, we examined the host defense responses to V. aestuarianus at the cellular and molecular levels, using flow-cytometry and real-time PCR techniques. We showed that hemocyte...
Tipo: Text Palavras-chave: Crassostrea gigas; Oyster; Pathogenesis; Vibrio aestuarianus; Bivalve immunity.
Ano: 2006 URL: http://archimer.ifremer.fr/doc/2006/publication-2182.pdf
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Défi MOREST - Mortalités etivales de l'huître creuse Crassostrea gigas - Caractérisation des facteurs de risques associés aux mortalités estivales - Synthèse du thème 4B - Caractérisation phénotypique des souches R Résistantes et S Susceptibles à la mortalité estivale. (La Rochelle 14-15 mars 2006) ArchiMer
Huvet, Arnaud; Royer, J.; Moal, J.; Burgeot, Thierry; Lapègue, Sylvie; Boulo, Viviane; Nicolas, Jean-louis; Lambert, C..
Les résultats du programme MOREST démontrent l'existence d'une composante génétique à la survie estivale du naissain et nous permettent de disposer aujourd'hui d'huTtres « résistantes» (R) et « susceptibles» (S) aux mortalités estivales (Dégremont et aL, 2002, 2005). La forte héritabilité estimée face à un tel système multifactoriel permet de suspecter que le caractère sélectionné puisse être gouverné par un ou un trés petit nombre de gènes sur une région chromosomique donnée. D'après le modèle d'interaction suspecté entre l'environnement, l'huître et des pathogènes opportunistes, une hiérarchie des facteurs, conduisant aux mortalités, permet de montrer qu'aucun d'entre eux ne peut provoquer à lui seul les mortalités mais que leur combinaison est...
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Ano: 2006 URL: http://archimer.ifremer.fr/doc/00306/41706/40917.pdf
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Differential distribution of V-type H+-ATPase and Na+/K+-ATPase in the branchial chamber of the palaemonid shrimp Macrobrachium amazonicum ArchiMer
Boudour-boucheker, Nesrine; Boulo, Viviane; Charmantier-daures, Mireille; Grousset, Evelyse; Anger, Klaus; Charmantier, Guy; Lorin-nebel, Catherine.
V-H+-ATPase and Na+/K+-ATPase were localized in the gills and branchiostegites of M. amazonicum and the effects of salinity on the branchial chamber ultrastructure and on the localization of transporters were investigated. Gills present septal and pillar cells. In freshwater (FW), the apical surface of pillar cells is amplified by extensive evaginations associated with mitochondria. V-H+-ATPase immunofluorescence was localized in the membranes of the apical evaginations and in clustered subapical areas of pillar cells, suggesting labeling of intracellular vesicle membranes. Na+/K+-ATPase labeling was restricted to the septal cells. No difference in immunostaining was recorded for both proteins according to salinity (FW vs. 25 PSU). In the branchiostegite,...
Tipo: Text Palavras-chave: Osmoregulation; Na+/K+-ATPase; V-type H+-ATPase; Gills; Branchiostegite.
Ano: 2014 URL: http://archimer.ifremer.fr/doc/00202/31352/37901.pdf
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Does the prevalence of Bonamia and Marteilia diseases be reduced on flat oysters (Ostrea edulis) of Atlantic and Mediterranean origin, when they are reared together with the Japanese oyster (Crassostrea gigas) in tidal ponds? ArchiMer
Bodoy, Alain; Bougrier, Serge; Geairon, Philippe; Garnier, Jacqueline; Boulo, Viviane; Heurtebise, Serge.
The flat oyster Ostrea edulis was originally reared in several regions of the European waters. The tidal saline ponds on the Atlantic coasts of France were highly productive before being hit by two consecutive epizooties. Parasitic diseases were described on this species, both of them leading 10 economic losses, as heavy mortalities have usually been recorded before the oysters reach the commercial size. Repeated experiments showed that the two diseases, Marteilia refringens and Bonamia ostreae are still active in this environment. Recently, some evidences were given that haemocytoblasts of Crassostrea gigas had the ability to lyze the Bonamia cells. Therefore, in an experiment of contiguous culture of the two species of oysters, the growth, mortality and...
Tipo: Text Palavras-chave: Molluscs; Mortalities; Growth; Pathology; Crassostra gigas; Ostrea edulis; Marteilia refringens; Bonamia ostreae.
Ano: 1991 URL: http://archimer.ifremer.fr/doc/1991/acte-3049.pdf
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Effects of fasting and re-alimentation on gill and intestinal morphology and indicators of osmoregulatory capacity in genetically selected sea bass (Dicentrarchus labrax) populations with contrasting tolerance to fasting ArchiMer
Alix, Maud; Blondeau-bidet, Eva; Grousset, Evelyse; Shiranghi, Ainaz; Vergnet, Alain; Guinand, Bruno; Chatain, Beatrice; Boulo, Viviane; Lignot, Jehan-herve.
Fasting and refeeding occur naturally in predators but this is largely ignored when dealing with farmed fish. Therefore, the effects of 3-week fasting and re-alimentation (2.5% of the individual body mass) were investigated using two genetically selected populations (F2 generation) of 250 g juvenile sea bass (Dicentrarchus labrax L.). Blood osmolarity, gill and intestinal morphology and expression of the sodium pump (Na+, K+-ATPase, NKA) were studied on two phenotypes showing different degrees of body mass loss during food deprivation: one group losing body mass rapidly during fasting (F +) and the other one limiting body mass loss during the same period (F-). Blood osmotic pressure significantly decreases due to re-alimentation in both groups, but this...
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Ano: 2017 URL: http://archimer.ifremer.fr/doc/00354/46521/46285.pdf
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Expression of immune-related genes in the oyster Crassostrea gigas during ontogenesis ArchiMer
Tirape, Ana; Bacque, Coralie; Brizard, Raphael; Vandenbulcke, Franck; Boulo, Viviane.
The work presented here reports the expression of immune-related genes during ontogenesis in the oyster Crassostrea gigar. Expression patterns of 18 selected genes showed that RNAs detected in oocytes and 2-4 cell embryos are of maternal origin and that gene transcription starts early after fertilization. The expression patterns of 4 genes (Cg-timp, Cg-tal, Cg-EcSOD and Drac3) suggested that hemocytes appear in the gastrula-trochophore stages. The localization of Cg-tal expression suggested that hematopoietic cells were derived from vessels and/or artery endothelia cells. Moreover, a bacterial challenge affected the level of expression of genes. Indeed, a change in expression levels was observed for Cg-LBP/BPI, Cg-timp, Drac3 and Cg-MyD88 genes in larval...
Tipo: Text Palavras-chave: Rel/NF kappa B pathway; Cell proliferation; Hematopoiesis; Mollusk immunity; Hemocytes; Bivalve mollusk.
Ano: 2007 URL: http://archimer.ifremer.fr/doc/2007/publication-2612.pdf
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First evidence of the activation of Cg-timp, an immune response component of pacific oysters, through a damage-associated molecular pattern pathway ArchiMer
Montagnani, Caroline; Avarre, Jean-christophe; De Lorgeril, Julien; Quiquand, M; Boulo, Viviane; Escoubas, Jean-michel.
In a previous work, we characterized a Crassostrea gigas cPNA (Cg-timp) encoding a protein which presents all the features of vertebrate tissue inhibitor of metalloproteinase (TIMP). The expression pattern of this gene led us to propose that Cg-timp is an important factor in oyster wound heating and defense mechanisms. Here we describe the analysis of Cg-timp expression in oysters challenged by live or dead bacteria as well as by bacterial secretory/excretory products and metalloproteinase. Surprisingly, bacterial secretory/excretory products activate Cg-timp gene expression whereas heat-inactivated ones do not. To address the question of the signal transduction pathway involved in Cg-timp gene activation, we isolated and sequenced Cg-timp promoter and...
Tipo: Text Palavras-chave: Damage associated molecular pattern; Tissue inhibitor of metalloproteinase; Metalloproteinase; Mollusk immunity; Crassostrea gigas; Oyster.
Ano: 2007 URL: http://archimer.ifremer.fr/doc/2007/publication-2094.pdf
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Generation and analysis of a 29,745 unique Expressed Sequence Tags from the Pacific oyster (Crassostrea gigas) assembled into a publicly accessible database: the GigasDatabase ArchiMer
Fleury, Elodie; Huvet, Arnaud; Lelong, Christophe; De Lorgeril, Julien; Boulo, Viviane; Gueguen, Yannick; Bachere, Evelyne; Tanguy, Arnaud; Moraga, Dario; Fabioux, Caroline; Lindeque, Penelope; Shaw, Jenny; Reinhardt, Richard; Prunet, Patrick; Davey, Grace; Lapegue, Sylvie; Sauvage, Christopher; Corporeau, Charlotte; Moal, Jeanne; Gavory, Frederick; Wincker, Patrick; Moreews, Francois; Klopp, Christophe; Mathieu, Michel; Boudry, Pierre; Favrel, Pascal.
Background: Although bivalves are among the most-studied marine organisms because of their ecological role and economic importance, very little information is available on the genome sequences of oyster species. This report documents three large-scale cDNA sequencing projects for the Pacific oyster Crassostrea gigas initiated to provide a large number of expressed sequence tags that were subsequently compiled in a publicly accessible database. This resource allowed for the identification of a large number of transcripts and provides valuable information for ongoing investigations of tissue-specific and stimulus-dependant gene expression patterns. These data are crucial for constructing comprehensive DNA microarrays, identifying single nucleotide...
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Ano: 2009 URL: http://archimer.ifremer.fr/doc/2009/publication-6770.pdf
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Genetically based resistance to summer mortality in the Pacific oyster (Crassostrea gigas) and its relationship with physiological, immunological characteristics and infection processes ArchiMer
Samain, Jean-francois; Degremont, Lionel; Soletchnik, Patrick; Haure, Joel; Bedier, Edouard; Ropert, Michel; Moal, Jeanne; Huvet, Arnaud; Bacca, Helene; Van Wormhoudt, A; Delaporte, Maryse; Costil, K; Pouvreau, Stephane; Lambert, Christian; Boulo, Viviane; Soudant, Philippe; Nicolas, Jean-louis; Leroux, Frederique; Renault, Tristan; Gagnaire, Beatrice; Geret, Florence; Boutet, Isabelle; Burgeot, Thierry; Boudry, Pierre.
Summer mortality of Pacific oysters is known in several countries. However no specific pathogen has been systematically associated with this phenomenon. A complex combination of environmental and biological parameters has been suggested as the cause and is now starting to be identified. A high genetic basis was found for survival in oysters when a first generation (G1) was tested in three sites during summer. This paper presents a synthesis on physiological characteristics of two selected groups ('R' and 'S', from families selected for resistance and susceptibility to summer mortality respectively), of the second and third generations. R and S showed improvement or reduction of survival compared with the control in both field and laboratory trials...
Tipo: Text Palavras-chave: ROS; Temperature; Stress; Hemocytes; Reproduction; Genetic selection; Resistant oyster; Crassostrea gigas; Pacific Oyster; Summer mortality.
Ano: 2007 URL: http://archimer.ifremer.fr/doc/2007/publication-2809.pdf
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Histological study of a cellular reaction in Ruditpaes decussatus infected by a protozoan ArchiMer
Chagot, Dominique; Comps, Michel; Boulo, Viviane; Ruano, Francisco; Grizel, Henri.
An infection caused by a Perkinsus -like parasite has been observed in Ruditapes decussatus from Portugal (Comps and Chagot, 1987). Following a pathological study carried out on clams affected by mortalities in the Algarve area, this parasite was found again, inducing a reaction process in the host. Histological examination reveals the parasite in the connective tissue of different organs. The adjoining tissues, particularly the epithelia, are not affected.
Tipo: Text Palavras-chave: Cells; Histology; Pathology; Ruditapes decussatus.
Ano: 1987 URL: http://archimer.ifremer.fr/doc/1987/acte-3109.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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Microarray-Based Identification of Gonad Transcripts Differentially Expressed Between Lines of Pacific Oyster Selected to Be Resistant or Susceptible to Summer Mortality ArchiMer
Fleury, Elodie; Moal, Jeanne; Boulo, Viviane; Daniel, Jean-yves; Mazurais, David; Henaut, Alain; Corporeau, Charlotte; Boudry, Pierre; Favrel, Pascal; Huvet, Arnaud.
Summer mortality of the Pacific oyster Crassostrea gigas is the result of a complex interaction between oysters, their environment, and pathogens. Heredity appears to be a major factor determining the sensitivity of oysters to summer mortality, allowing resistant (R) and susceptible (S) lines to be produced. We conducted genome-wide expression profiling of R and S gonads during the 3-month period preceding a summer mortality event, using a cDNA microarray that we designed. ANOVA analysis revealed that 34 genes were differentially expressed between R and S lines on four dates preceding the mortality event. Annotation of some of these genes highlights reproduction and its allocation and antioxidant defenses as the main pathways that operate differentially...
Tipo: Text Palavras-chave: Mollusca; Bivalvia; Gene expression; CDNA microarrays; Reproduction; Oxidative stress; Energy metabolism.
Ano: 2010 URL: http://archimer.ifremer.fr/doc/00005/11590/8258.pdf
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Monoclonal antibodies : a tool for molluscan pathology ArchiMer
Mialhe, Eric; Boulo, Viviane; Grizel, Henri; Rogier, Hervé; Paolucci, Francis.
Hybridoma technology is reviewed, and the characteristics of monoclonal antibodies are compared with those of polyclonal antibodies. The contribution of monoclonal antibodies to molluscan pathology is developed with special emphasis on their use as diagnostic tools. The results of studies with monoclonal antibodies prepared against the protozoan oyster pathogen Bonamia ostreae are briefly described.
Tipo: Text Palavras-chave: Pathology; Molluscs; Monoclonal antibody technology.
Ano: 1988 URL: http://archimer.ifremer.fr/doc/1988/publication-3121.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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Monoclonal antibodies against sporangia and spores of Marteilia sp. (Protozoa: Ascetospora) ArchiMer
Robledo, Jaf; Boulo, Viviane; Mialhe, Eric; Despres, Béatrice; Figueras, Antonio.
Digestive glands of mussels Mytilus edulis from Brittany, France, infected with Marteilia sp. (Ascetospora) were used to purify the parasite. A modification of a previously used purification protocol increased purification efficiency, permitting sporangial primordia and sporangia of Marteilia sp. to be obtained. Mouse (Balb/c) monoclonal antibodies were generated against this parasite. From the fusion, 26 monoclonal antibodies against Marteilia sp. were obtained. Antibodies from 6 clones reacted only with Marteilia sp. cells and not with normal host tissues. Four of these antibodies (1/1-3, 3/1-1, 4/1-1 and 6/2-3) reacted with the sporangia wall and two with the spore cytoplasm (9/1-1 and 12/5-1). Antibodies cross-reacted with Marteilia refringens from...
Tipo: Text Palavras-chave: Marteilia sp.; Mytilus edulis; Monoclonal antibodies.
Ano: 1994 URL: http://archimer.ifremer.fr/doc/00196/30730/29404.pdf
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