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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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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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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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The ClC-3 chloride channel and osmoregulation in the European Sea Bass, Dicentrarchus labrax ArchiMer
Bossus, Maryline; Charmantier, Guy; Blondeau-bidet, Eva; Valletta, Bianca; Boulo, Viviane; Lorin-nebel, Catherine.
Dicentrarchus labrax migrates between sea (SW), brackish and fresh water (FW) where chloride concentrations and requirements for chloride handling change: in FW, fish absorb chloride and restrict renal losses; in SW, they excrete chloride. In this study, the expression and localization of ClC-3 and Na+/K+-ATPase (NKA) were studied in fish adapted to SW, or exposed to FW from 10 min to 30 days. In gills, NKA-alpha 1 subunit expression transiently increased from 10 min and reached a stabilized intermediate expression level after 24 h in FW. ClC-3 co-localized with NKA in the basolateral membrane of mitochondria-rich cells (MRCs) at all conditions. The intensity of MRC ClC-3 immunostaining was significantly higher (by 50 %) 1 h after the transfer to FW,...
Tipo: Text Palavras-chave: ClC-3 chloride channel; Na+/K+-ATPase; Dicentrarchus labrax; Osmoregulation; Osmosensing.
Ano: 2013 URL: http://archimer.ifremer.fr/doc/00149/26044/25185.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
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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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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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Salinity tolerance, ontogeny of osmoregulation and zootechnical improvement in the larval rearing of the Caledonian Blue Shrimp, Litopenaeus stylirostris (Decapoda, Penaeidae) ArchiMer
Pham, Dominique; Charmantier, Guy; Wabete, Nelly; Boulo, Viviane; Broutoi, Francis; Mailliez, Jean-rene; Peignon, Jean-marie; Charmantier-daures, Mireille.
The ontogeny of osmoregulationwas investigated in Litopenaeus stylirostris by studying salinity tolerance and osmoregulatory capacity. Shrimp at different larval and postlarval stages were exposed to various salinities and survival was monitored for 24 h. Survival rates exceeded 80% at salinity over 25 ppt (750 mOsm.kg(-1)) at all the stages. At salinities below to 25 ppt, salinity tolerance was higher in nauplii and zoeae than in mysis larvae. Postlarval stages were able to withstand lower salinities, e.g. 6.0 ppt (176 mOsm.kg(-1)) at PL9 stage, but they were more sensitive than larvae to salinities over 35 ppt (1035 mOsm.kg(-1)). Zoea and mysis larvae slightly hyper-regulated at all tested salinities. After metamorphosis, postlarvae progressively...
Tipo: Text Palavras-chave: Penaeid; Litopenaeus stylirostris; Ontogeny; Osmoregulation; Growth.
Ano: 2012 URL: http://archimer.ifremer.fr/doc/00107/21843/20058.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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Osmoregulation in larvae and juveniles of two recently separated Macrobrachium species: Expression patterns of ion transporter genes ArchiMer
Boudour-boucheker, Nesrine; Boulo, Viviane; Charmantier-daures, Mireille; Anger, Klaus; Charmantier, Guy; Lorin-nebel, Catherine.
In this comparative study, osmoregulatory mechanisms were analyzed in two closely related species of palaemonid shrimp from Brazil, Macrobrachium pantanalense and Macrobrachium amazonicum. A previous investigation showed that all postembryonic stages of M. pantanalense from inland waters of the Pantanal are able to hyper-osmoregulate in fresh water, while this species was not able to hypo-osmoregulate at high salinities. In M. amazonicum originating from the Amazon estuary, in contrast, all stages are able to hypo-osmoregulate, but only first-stage larvae, late juveniles and adults are able to hyper-osmoregulate in fresh water. The underlying molecular mechanisms of these physiological differences have not been known. We therefore investigated the...
Tipo: Text Palavras-chave: Salinity; Crustaceans; Branchial chamber; NHE3 transporter; V-type H+-ATPase; Na+/K+-ATPase.
Ano: 2016 URL: http://archimer.ifremer.fr/doc/00313/42427/41899.pdf
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Monoclonal antibody against a gill Rickettsiales-like organism of Pecten maxiumus (Bivalvia): application to indirect immunofluorescence diagnosis ArchiMer
Le Gall, Gyslaine; Mourton, Chantal; Boulo, Viviane; Paolucci, Francis; Pau, Bernard; Mialhe, Eric.
Using hybridoma technology, monoclonal antibodies (MABs) were prepared against a Rickettsiales-like organism (RLO) from the gills of the Saint-Jacques scallop Pecten maximus. The 5D7 MAB was then selected as a specific reagent suitable for an indirect immunofluorescence assay (IIFA) and evaluated for RLO diagnosis on P. maxiumus gill smears. Monoclonal antibody 5D7 showed no cross-reactivity with an RLO associated with a related pectinid species Chlamys opercularis
Tipo: Text Palavras-chave: Diagnosis; Rickettsiales like organism; Monoclona antibody; Pecten maximus.
Ano: 1992 URL: http://archimer.ifremer.fr/doc/1992/publication-2762.pdf
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Phénomène des branchies orange dans les élevages de crevettes en Nouvelle-Calédonie ArchiMer
Lemonnier, Hugues; Barri, Kiam; Pham, Dominique; Wabete, Nelly; Boulo, Viviane; Lignot, Jean-herve; Royer, Florence; Hubert, Morgane; Blockmans, Bernard; Pierrot, Thomas; Laugier, Thierry; Tostin, Nathalie.
L'aquaculture est une activité importante du paysage économique de la Nouvelle-Calédonie., La crevette Bleue du Pacifique, Litopenaeus stylirostris est aujourd’hui l’espèce cible, élevée en semi-intensif, dans des bassins de terre de 4 à 10 hectares. Une coloration orange prononcée au niveau des branchies, comparable à celle observée suite à un stress hypoxique alors qu’aucun déficit en oxygène n’est constaté, est apparue récemment engendrant une décote du produit à la vente et donc un risque économique supplémentaire pour cette filière déjà fragilisée par des mortalités à vibrioses. Le phénomène se déclare à un poids moyen des animaux compris entre 10 et 18 g pour une durée moyenne d’élevage de 70 jours. L'intensité de la couleur évolue avec le stade de...
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Ano: 2014 URL: http://archimer.ifremer.fr/doc/00248/35909/34462.pdf
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Osmoregulation pattern and salinity tolerance of the white shrimp Litopenaeus vannamei (Boone, 1931) during post-embryonic development ArchiMer
Chong-robles, Jennyfers; Charmantier, Guy; Boulo, Viviane; Lizarraga-valdez, Joel; Enriquez-paredes, Luis M.; Giffard-mena, Ivone.
The euryhaline white shrimp Litopenaeus vannamei lives in both coastal and oceanic areas through ontogeny. Its osmoregulation pattern and variations in its tolerance to salinity are partially known from several studies under different experimental conditions (developmental stages, salinities and acclimation procedures). Although L. vannamei is recognized as one of the most euryhaline penaeid species, with adults and juveniles exhibiting an hyper-hypo-osmoregulatory pattern and being able to tolerate a wide salinity range, little is known on larval and early postlarval strategies to cope with salinity fluctuations. In order to establish their euryhalinity range and to fully understand the ontogenetic changes in L. vannamei osmoregulatory pattern, we...
Tipo: Text Palavras-chave: Osmotic pressure; Hypo-hyper osmoregulation; Osmotic stress; Survival; Crustacean; Larvae.
Ano: 2014 URL: http://archimer.ifremer.fr/doc/00169/28068/26283.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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Recent advances in EST sequencing in Crassostrea gigas: towards the sequencing of the Pacific oyster genome ArchiMer
Boudry, Pierre; Huvet, Arnaud; Lapegue, Sylvie; Sauvage, Christopher; Tanguy, A.; Moraga, D.; Goostrey, A.; Prunet, P.; Boulo, Viviane; Gueguen, Y.; Moal, Jeanne; Fleury, Elodie; Samain, Jean-francois; Mathieu, M.; Favrel, P..
Rationale for EST and genome sequencing of the Pacific oyster: • its membership of the Lophotrochozoa, an little studied branch of the Eukaryotes, • its worldwide aquacultural importance (4.2 million metric tons, worth 3.5 billion US dollars), • its high fecundity, with concomitantly high DNA polymorphism, • its use as a biosensor of coastal pollution. In 2004, an international community of biologists, the Oyster Genome Consortium (OGC) led by D. Hedgecock (University of Southern California) and uniting 70 participants from 10 countries, first presented the Pacific oyster Crassostrea gigas as a genome-sequencing candidate to Joint Genome Institute (JGI, USA) (Hedgecock et al., 2005). Besides this project (that remains to be accepted), several EST...
Tipo: Text Palavras-chave: Characteristics; Genome size; Genome; Crassostrea gigas; Oyster; Genetic.
Ano: 2007 URL: http://archimer.ifremer.fr/doc/2007/acte-3414.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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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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Ontogeny of osmoregulation in the Pacific blue shrimp, Litopenaeus stylirostris (decapoda, penaeidae): Deciphering the role of the Na+/K+-ATPase ArchiMer
Pham, Dominique; Charmantier, Guy; Boulo, Viviane; Wabete, Nelly; Ansquer, Dominique; Dauga, Clement; Grousset, Evelyse; Labreuche, Yannick; Charmantier-daures, Mireille.
The role of the main ion transporting enzyme Na +/K + − ATPase in osmoregulation processes was investigated in Litopenaeus stylirostris. The development and localization of the osmoregulation sites were studied during ontogenesis by immunodetection of Na+ K+-ATPase using monoclonal antibodies and transmission electron microscopy (TEM). Osmoregulation sites were identified as the pleurae and branchiostegites in the zoeae and mysis stages. In the subsequent post-metamorphic stages the osmoregulatory function was mainly located in the epipodites and branchiostegites and osmotic regulation was later detected in the gills. The presence of ionocytes and microvilli in these tissues confirmed their role in ionic processes. The complete open reading frame of the...
Tipo: Text Palavras-chave: Penaeid; Litopenaeus stylirostris; Ontogeny; Osmoregulation; Na+/K+-ATPase; RNA interference.
Ano: 2016 URL: http://archimer.ifremer.fr/doc/00311/42176/41477.pdf
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