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Rapid and sensitive detection of ostreid Herpesvirus 1 in oyster samples by real-time PCR (Poster) ArchiMer
Pepin, Jean-francois; Riou, Antoine; Renault, Tristan.
Herpes and herpes-like virus infections have been reported in various marine mollusc species associated with high mortality rates. Following the characterisation and genome sequencing of ostreid herpesvirus 1 (OsHV-1), specific diagnostic tools have been developed based on conventional PCR techniques or in situ hybridisation. We have now developed a real-time PCR assay for rapid, sensitive and quantitative detection of OsHV-1, and compared it with a conventional PCR technique described previously. The new assay utilised SYBR® Green chemistry with specific primers C9/C10 targeting the C region, which is present in two copies in the OsHV-1 genome and encodes two proteins of unknown function. The assay was applied successfully to rapid diagnosis (100 minutes)...
Tipo: Text Palavras-chave: Crassostrea gigas; PCR; Oyster; OsHV 1; Detection; Herpes virus 1.
Ano: 2007 URL: http://archimer.ifremer.fr/doc/2007/acte-3653.pdf
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Dual transcriptomics of virus-host interactions: comparing two Pacific oyster families presenting contrasted susceptibility to ostreid herpesvirus 1 ArchiMer
Segarra, Amelie; Mauduit, Florian; Faury, Nicole; Trancart, Suzanne; Degremont, Lionel; Tourbiez, Delphine; Haffner, Philippe; Barbosa-solomieu, Valerie; Pepin, Jean-francois; Travers, Marie-agnes; Renault, Tristan.
Background: Massive mortality outbreaks affecting Pacific oyster (Crassostrea gigas) spat in various countries have been associated with the detection of a herpesvirus called ostreid herpesvirus type 1 (OsHV-1). However, few studies have been performed to understand and follow viral gene expression, as it has been done in vertebrate herpesviruses. In this work, experimental infection trials of C. gigas spat with OsHV-1 were conducted in order to test the susceptibility of several bi-parental oyster families to this virus and to analyze host-pathogen interactions using in vivo transcriptomic approaches. Results: The divergent response of these oyster families in terms of mortality confirmed that susceptibility to OsHV-1 infection has a significant...
Tipo: Text Palavras-chave: Interactions; Ostreid herpesvirus 1; Gene expression; Susceptibility; Crassostrea gigas.
Ano: 2014 URL: http://archimer.ifremer.fr/doc/00199/31029/29431.pdf
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Recherche et caractérisation d'immunoglobulines dans les oeufs du loup (Dicentrarchus labrax) ArchiMer
Pepin, Jean-francois; Breuil, Gilles; Rigobert, P; Romestand, Bernard.
The occurrence of maternal immunity transfer in Sea bass (Dicentrarchus labrax) was investigated using ELISA and Western blot technics. In order to determine the possible presence of immunoglobulin (Ig), egg proteins obtained from fertilized eggs were analyzed. After several extraction protocol assays (precipitation, chromatography), monoclonal and polyclonal antibody probes specific to Sea bass Ig allowed to confirm the presence of an immunoglobulin-like protein in eggs with very low level. Biochemical characterization was carried out using SDS-PAGE. Results indicated that egg Ig had a molecular weight of 206 kDa. They were smaller than seric IgM (800 kDa). The heavy (H) and light (L) chains of the egg Ig and of the seric IgM had similar molecular weight,...
Tipo: Text Palavras-chave: Immunoglobuline; Transfert immunité maternelle; ELISA; Œuf; Poisson; Immunoglobulin; Maternal immunity transfer; ELISA; Eggs; Fish.
Ano: 1997 URL: http://archimer.ifremer.fr/doc/00272/38275/36458.pdf
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Microbial balance, and triggering of recent mussel mortality events in France: Is there a link? ArchiMer
Guesdon, Stephane; Travers, Marie-agnes; Hervio Heath, Dominique; Derrien, Annick; Genauzeau, Sylvie; Schmitt, Anne; Quintric, Laure; Leroi, Laura; Pepin, Jean-francois.
Since the spring of 2014, the mussel farming of Charente-Maritime and Vendée (French Atlantic coast) has experienced an unprecedented crisis. Mass mortality events of blue mussel (M. edulis) have affected livestock areas, decimating up to 100% of production in some farming areas. Investigation for infectious agents in moribund mussels revealed presence of bacteria capable of inducing mortalities. In this context, the MORBLEU research project (DPMA-Ifremer agreement) was initiated to explore potentially aggravating factors, associated or correlated with mortality of mussels. The aim of the present study is to support the hypothesis that these mortality episodes can be linked to an evolution of microbial communities. Based on historical data from Ifremer...
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Ano: 2017 URL: http://archimer.ifremer.fr/doc/00415/52696/53566.pdf
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Bilan 2015 du dispositif national de surveillance de la santé des mollusques marins ArchiMer
Lupo, Coralie; Osta Amigo, Axel; Fleury, Elodie; Robert, Stephane; Garcia, Celine; Arzul, Isabelle; Baillon, Laury; Bechemin, Christian; Canier, Lydie; Chollet, Bruno; Dechamps, Lucie; Dubreuil, Christine; Faury, Nicole; Francois, Cyrille; Godfrin, Yoann; Lapegue, Sylvie; Morga, Benjamin; Travers, Marie-agnes; Tourbiez, Delphine; Masson, Jean-claude; Verin, Francoise; Cordier, Remy; Gangnery, Aline; Louis, Wilfried; Mary, Charlotte; Normand, Julien; Penot, Julia; Cheve, Julien; Dagault, Francoise; Le Jolivet, Aurore; Le Gal, Dominique; Lebrun, Luc; Bellec, Gwenael; Bouget, Jean-francois; Cochennec-laureau, Nathalie; Palvadeau, Hubert; Grizon, James; Chabirand, Jean-michel; Pepin, Jean-francois; Seugnet, Jean-luc; D'Amico, Florence; Maurer, Daniele; Le Gall, Patrik; Mortreux, Serge; Baldi, Yoann; Orsoni, Valerie; Bouchoucha, Marc; Le Roy, Valerian; Pouvreau, Stephane; Queau, Isabelle; Lamoureux, Alice.
Depuis 1992, la surveillance de la santé des mollusques marins du littoral français est assurée par le réseau de Pathologie des Mollusques (Repamo). Ses activités s’inscrivent dans le cadre de la Directive Européenne 2006/88/CE. Depuis son évaluation par la plateforme nationale d’épidémiosurveillance en santé animale en 2012, l’objectif de surveillance est la détection précoce des infections dues à des organismes pathogènes exotiques et émergents affectant les mollusques marins sauvages et d’élevage. L’année 2015 est la première année de transition pour laquelle un début d’évolution des modalités de surveillance de la santé des mollusques marins animées par l’Ifremer a été amorcé. Un dispositif hybride de surveillance a été mis en place, s’appuyant sur...
Tipo: Text Palavras-chave: Réseau; Surveillance; Pathologie; Mollusques; Coquillages; Santé; Mortalité; Maladie.
Ano: 2016 URL: http://archimer.ifremer.fr/doc/00324/43486/42945.pdf
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Ostreid herpes virus 1 infection in families of the Pacific oyster, Crassostrea gigas, during a summer mortality outbreak: Differences in viral DNA detection and quantification using real-time PCR ArchiMer
Sauvage, Christopher; Pepin, Jean-francois; Lapegue, Sylvie; Boudry, Pierre; Renault, Tristan.
Ostreid herpes virus 1 (OsHV-1) infections, notably reported in Europe and the USA, are closely associated with significant mortalities of the Pacific oyster, Crassostrea gigas, especially during its early stages of life. In summer 2006, we monitored mortality by strict daily verification of three full-sib families of oysters reared under common conditions. We quantified OsHV-1 using real-time PCR in dead and living individuals during and after a mortality event. Mortality events were severe and brief, but significantly different between tested families (cumulative mortality ranging from 1.2 to 49%). Real-time PCR assays revealed different viral DNA loads in dead individuals from different families (P < 0.001). Moreover, the mean level of infection...
Tipo: Text Palavras-chave: Viral DNA quantification; Real time PCR; OsHV 1; Mortality; Crassostrea gigas.
Ano: 2009 URL: http://archimer.ifremer.fr/doc/2009/publication-6265.pdf
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Mise au point d'outils de gestion des gamètes mâles d'huîtres creuses Crassostrea gigas diploïdes et tétraploïdes à des fins de conservation et de diffusion ArchiMer
Brizard, Raphael; Boudry, Pierre; Haffray, Patrick; Labbe, C.; Suquet, Marc; Robert, Rene; Masse, G.; Pepin, Jean-francois; Saulnier, Denis.
In order to improve oyster culture, many research programs try and propose new methods aiming at improving genetically the product performances and quality (selection, polyploidisation). As for terrestrial animal production, the continuity of the different works as well as the spreading of the genetical progress would be made easier by the development of conservation techniques of the selected genotypes. Cryopreservation, which is often used in selection programs of terrestrial animals or fish farming species, is not always available for bivalves. This technique would enable the conservation of the gametes of individuals selected for their high growth potential, their resistance to diseases and/or their quality for a smaller cost and less risks than the...
Tipo: Text Palavras-chave: Tetraploïde; Diploïde; Concervation; Spermatozoides; Cryoconservation; Génétique; Crassostrea gigas; Huîtres.
Ano: 2006 URL: http://archimer.ifremer.fr/doc/2006/acte-3339.pdf
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Défi MOREST - Mortalités estivales de l'huître creuse Crassostrea gigas - Caractérisation des facteurs de risques associés aux mortalités estivales - Synthèse du thème 5 - Risque associé aux pathogènes. (La Rochelle 14-15 mars 2006) ArchiMer
Nicolas, Jean-louis; Renault, Tristan; Gagnaire, Beatrice; Garcia, Celine; Le Roux, F; Miossec, Laurence; Pepin, Jean-francois; Saulnier, Denis.
L'un des objectifs principaux des travaux en pathologie des mollusques est d'identifier et d'étudier des agents infectieux capables d'induire des maladies et des mortalités chez les coquillages, en particulier, chez I'huître creuse, Crassostrea gigas, espèce principalement exploitée en France. Ces travaux semblent d'autant plus indispensables qu'il n'existait pas, jusqu'à ces dernières années, de maladies identifiées chez I'huître creuse en Europe excepté le virus herpès suspecté dès 1995 de provoquer des mortalités de naissain (Renault et al, 1995). Les principales maladies des espèces proches sont dues à des parasites comme Haplosporidium nelsoni (Ford et Heskin, 1987) et Perkinsus marinus Olivier et al, 1 998) pour Crassostrea virginica, Marteilia...
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Ano: 2006 URL: http://archimer.ifremer.fr/doc/00304/41535/40743.pdf
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Summer mortality outbreaks of French Pacific oysters Crassostrea gigas in 2008 and 2009 ArchiMer
Renault, Tristan; Allain, Gwenhael; Arzul, Isabelle; Chollet, Bruno; Cobret, Laetitia; De Decker, Sophie; Faury, Nicole; Ferrand, Sylvie; Francois, Cyrille; Garcia, Celine; Haffner, Philippe; Joly, Jean-pierre; Miossec, Laurence; Morga, Benjamin; Nicolas, Jean-louis; Omnes, Emmanuelle; Pepin, Jean-francois; Saulnier, Denis; Schikorski, David; Segarra, Amelie.
Abnormal mortality outbreaks occurred in the majority of Pacific oyster (Crassostrea gigas) producing areas in France in 2008 during the summer period. These outbreaks were usually sudden and severe (up to 100%) and affected essentially spat (oysters less than 1 year-old) and juveniles (12 to 18 month-old oysters). Crassostrea gigas was the single affected species Mortality outbreaks were first observed from mid May to mid June in Brittany, Vendée, Charente Maritime and Mediterranean coast A second episode of massive mortality events was reported at the end of June and the beginning of July. All oyster production sites were affected except some particular locations including Arcachon Bay and Brittany. Mortality occurred latter in Arcachon Bay in August....
Tipo: Text Palavras-chave: Abnormal mortality; Oysters; Crassostrea gigas; Pathogens.
Ano: 2009 URL: http://archimer.ifremer.fr/doc/00014/12554/9420.pdf
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Projet de recherche "MORBLEU". Environnement des écosystèmes mytilicoles et crises de mortalité récentes, quelles interactions? Etude des facteurs pouvant favoriser les surmortalités de moules bleues (Mytilus edulis) dans les pertuis charentais et en Vendée (2014-2016) ArchiMer
Pepin, Jean-francois; Travers, Marie-agnes.
Durant l’hiver 2014 jusqu’à mai 2014, des mortalités exceptionnelles (50-100%) ont affectées la moule bleue, Mytilus edulis dans les Pertuis Charentais (principalement secteur du Pertuis Breton) et en Vendée. A la demande du Ministère (DPMA) une expertise environnementale a été demandée à l’Ifremer. L’expertise a permis de montrer que les conditions hydrologiques de l’hiver 2014 ne permettent pas de mettre en évidence une cause environnementale à ces mortalités exceptionnelles. Les analyses en pathologie réalisées ont révélées dans les moules la présence de bactéries appartenant au groupe bactérien Vibrio splendidus, dont la virulence a été confirmée expérimentalement. Dans ce contexte, la DPMA a sollicité l’Ifremer pour conduire en 20105 une étude...
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Ano: 2016 URL: http://archimer.ifremer.fr/doc/00361/47174/47141.pdf
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Short Technical Report for OsHV-1 detection and quantification by Real Time Polynerase Chain Reaction using OsHV-1 DNA polymerase sequence ArchiMer
Pepin, Jean-francois.
The present report completes a previously described procedure routinely used for the detection and quantification of OsHV-1 by Sybr green Real Time Polymerase Chain Reaction targeting the OsHV-1 DNA polymerase sequence. More particularly, this document brings information about analytical sensitivity, quantitation limit, assay precision, reproducibility of the technique.
Tipo: Text Palavras-chave: PCR quantitative HDP-F/R; Virus OsHV-1; Diagnostic; DNA polymerase; Validation; Real-Time PCR HDP-F/R; OsHV-1; Diagnostic; DNA polymerase; Validation.
Ano: 2013 URL: http://archimer.ifremer.fr/doc/00137/24814/22900.pdf
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Development of a new diagnostic tool (Mini-Array) for ostreid Herpesvrus 1 (OsHV-1) detection in the Pacific oyster Crassostre gigas ArchiMer
Varsamos, S.; Pepin, Jean-francois; Sauvage, Christopher; Renault, Tristan.
OsHV-1 was the first herpesvirus to be identified in an invertebrate host and is associated to mortalities in the Pacific oyster (Crassostrea gigas) and other economically important bivalve species. Infections due to OsHV-1 are frequently reported and cause high mortality rates in larvae and spat of C. gigas. Detection of this pathogen is usually performed by PCR and/or in situ hybridization. This presentation will focus on the development of a new molecular diagnostic tool for OsHV-1 (and OsHV-1 variant 1) detection: the mini-array. This detection system, which allows multiple pathogens detection, combines viral DNA amplification by PCR and a specific hybridisation on a nylon membrane, revealed by a colorimetric reaction (DNA chip). Multiplex PCR involves...
Tipo: Text Palavras-chave: Diagnostic; Crassotrea gigas; Oyster; Detection; Herpesviridae; OsHV 1.
Ano: 2007 URL: http://archimer.ifremer.fr/doc/2007/acte-3652.pdf
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Experimental ostreid herpesvirus 1 infection of the Pacific oyster Crassostrea gigas: Kinetics of virus DNA detection by q-PCR in seawater and in oyster samples ArchiMer
Schikorski, David; Faury, Nicole; Pepin, Jean-francois; Saulnier, Denis; Tourbiez, Delphine; Renault, Tristan.
Herpes- and herpes-like viruses are known to infect a wide range of bivalve mollusc species throughout the world. Abnormal summer mortalities associated to the detection of ostreid herpesvirus 1 (OsHV-1) have been currently reported in France among larvae and spat of the Pacific cupped oyster Crassostrea gigas. In the present work, we have developed an experimental protocol of horizontal transmission based on the cohabitation between healthy and experimentally infected oysters. Through a cohabitation trial, the kinetics of OsHV-1 detection in different oyster organs and seawater samples were investigated and characterized for the first time using real time quantitative PCR.
Tipo: Text Palavras-chave: Ostreid herpesvirus 1; Cupped Pacific oyster; Crassostrea gigas; Experimental infection; Kinetic viral DNA quantification.
Ano: 2011 URL: http://archimer.ifremer.fr/doc/00025/13607/10688.pdf
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Rapport de synthèse sur la communication et la valorisation des données liées à l’étude MORBLEU et aux mortalités de moules bleues en France ArchiMer
Pepin, Jean-francois.
Ce rapport s’inscrit dans le cadre du projet MORBLEU mené par l’Ifremer entre 2015 et 2019 en réponse à une demande de la DPMA. Ce document présente une synthèse des éléments de communication et de valorisation des données et des connaissances, liées à l’étude MORBLEU et aux mortalités de moules bleues en France sur cette période.
Tipo: Text Palavras-chave: Restitution; Communication; Valorisation; Rapport de synthèse; Étude MORBLEU; Presse; Moules; Mytilus sp; Mortalité.
Ano: 2020 URL: https://archimer.ifremer.fr/doc/00668/77962/80173.pdf
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Compte rendu final du projet Triplofimer : Fiabilité et qualité de la production de naissains triploïdes d'huître creuse (Crassostrea gigas) ArchiMer
Suquet, Marc; Agogue, Julie; Brizard, Raphael; Haffray, Pierrick; Labbe, Catherine; Le Coz, Jean-rene; Pepin, Jean-francois; Rault, Paul; Saulnier, Denis.
Ce rapport fait état d'une étude réalisée au laboratoire de Génétique et Pathologie de l'IFREMER dans le cadre de l'analyse d'épisodes de mortalité anormale survenue en écloserie privée de crevettes pénéides. Plusieurs hypothèses ont été avancées. La présence de Vibrio harveyi a amené à constituer une collection de 40 souches à partir desquelles une identification bactériologique a été effectuée. Elle montre en effet une dominance de souches appartenant à un groupe homogène de Vibrio harveyi. La technique de ribotypage (digestion d'ADN génomique, transfert de Southem-blot et hybridation avec une sonde spécifique) est alors mise en place. Plusieurs difficultés techniques ont ralenti l'analyse et n'ont donc pas permis d'avoir des résultats assez avancés pour...
Tipo: Text Palavras-chave: Huître creuse; Crassostrea gigas; Tétraploïdes; Sperme; Conservation; Diagnostic zoo-sanitaire; Flore bactérienne; Herpès virus..
Ano: 2007 URL: http://archimer.ifremer.fr/doc/00304/41554/40762.pdf
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New Insight for the Genetic Evaluation of Resistance to Ostreid Herpesvirus Infection, a Worldwide Disease, in Crassostrea gigas ArchiMer
Degremont, Lionel; Lamy, Jean-baptiste; Pepin, Jean-francois; Travers, Marie-agnes; Renault, Tristan.
The Pacific oyster, Crassostrea gigas, is the most important commercial oyster species cultivated in the world. Meanwhile, the ostreid herpesvirus 1 (OsHV-1) is one of the major pathogens affecting the Pacific oyster, and numerous mortality outbreaks related to this pathogen are now reported worldwide. To assess the genetic basis of resistance to OsHV-1 infection in spat C. gigas and to facilitate breeding programs for such a trait, if any exist, we compared the mortality of half- and full-sib families using three field methods and a controlled challenge by OsHV-1 in the laboratory. In the field, three methods were tested: (A) one family per bag; (B) one family per small soft mesh bag and all families inside one bag; (C) same as the previous methods but...
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Ano: 2015 URL: http://archimer.ifremer.fr/doc/00269/38066/36178.pdf
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Recherches d'ADN d'hepès virus infectant les mollusques dans les populations d'huîtres en milieu naturel et dans leur environnement ArchiMer
Pepin, Jean-francois; Solliec, Gaelle; Vigneron, Vassilia; Thebault, Anne; Lamothe, Julien; Renault, Tristan.
The presence of a herpes-type virus has been demonstrated in various species of marine shellfish (bivalves and gastropoda) throughout different regions of the world (Europe, North and South America, Asia, Australia), and is frequently associated with high mortality levels. Nevertheless, apart from the sanitary monitoring in mollusc farming, very little data is available about the detection of this type of virus in the natural populations of bivalves and about the persistence of such viruses in sea water. Various diagnostic tools have been developed in order to detect the oyster herpes virus (Ostreid herpesvirus, OsHV). The presence of this virus has been looked for by PCR in marine water samples coming from fattening ponds, in order to precise the natural...
Tipo: Text Palavras-chave: Levures; Eau de mer; Diagnostic; Huîtres; Mollusques; OsHV; Herpèsvirus.
Ano: 2008 URL: http://archimer.ifremer.fr/doc/2008/acte-3716.pdf
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Experimental infection of Pacific oyster Crassostrea gigas spat by ostreid herpesvirus 1: demonstration of oyster spat susceptibility ArchiMer
Schikorski, David; Renault, Tristan; Saulnier, Denis; Faury, Nicole; Moreau, Pierrick; Pepin, Jean-francois.
In 2008 and 2009, acute mortalities occurred in France among Pacific cupped oyster, Crassostrea gigas, spat. Different hypothesis including the implication of environmental factors, toxic algae and/or pathogens have been explored. Diagnostic tests indicated that OsHV-1 including a particular genotype, termed OsHV-1 mu Var, was detected in most of samples and especially in moribund oysters with the highlighting of virus particles looking like herpes viruses by TEM examination. In this study, an experimental protocol to reproduce OsHV-1 infection in laboratory conditions was developed. This protocol was based on the intramuscular injection of filtered (0.22 mu m) tissue homogenates prepared from naturally OsHV-1 infected spat collected on French coasts...
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Ano: 2011 URL: http://archimer.ifremer.fr/doc/00037/14795/12104.pdf
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Mass mortalities of Pacific oysters Crassostrea gigas reflect infectious diseases and vary with farming practices in the Mediterranean Thau lagoon, France ArchiMer
Pernet, Fabrice; Barret, Jean; Le Gall, Patrik; Corporeau, Charlotte; Degremont, Lionel; Lagarde, Franck; Pepin, Jean-francois; Keck, Nicolas.
Experiments were conducted to investigate the effects of farming sites, seeding date, age of oysters, ploidy level and rearing structures on mortality, pathogens and energetic reserves of oysters Crassostrea gigas. Mortality of oysters coincided with infections involving, primarily, the ostreid herpesvirus OsHV-1 and, secondarily, bacteria of the group Vibrio splendidus, when seawater temperature was between 17 and 24°C. Mortality started in early May and lasted until late September, with an interruption in July and August when seawater temperatures rose above 24°C. Mortality varied with farming site: oysters maintained in the open Mediterranean Sea showed no mortality and OsHV-1 was occasionally detected at a low level only, while oysters maintained in...
Tipo: Text Palavras-chave: Pacific oyster; Mortality; Disease; Farming practices; Hydrodynamics; Energetic.
Ano: 2012 URL: http://archimer.ifremer.fr/doc/00086/19739/17379.pdf
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Mortalité du naissain d’Huître creuse Crassostrea gigas dans l’étang de Thau en 2009 ArchiMer
Pernet, Fabrice; Barret, Jean; Le Gall, Patrik; Malet, Nathalie; Pastoureaud, Annie; Munaron, Dominique; De Lorgeril, Julien; Bachere, Evelyne; Vaquer, Andre; Huvet, Arnaud; Corporeau, Charlotte; Normand, Julien; Boudry, Pierre; Moal, Jeanne; Quere, Claudie; Quilien, Virgile; Daniel, Jean-yves; Pepin, Jean-francois; Saulnier, Denis; Gonzalez, Jean-louis.
This study was conducted in the Mediterranean area during a mass mortality event of Pacific oyster Crassostrea gigas. The objectives of this work were to increase our understanding of the mass mortality phenomena in Pacific oysters, to identify risky cultivation practises and to propose mitigation strategies. In the first part of this work, we followed environmental parameters of the Thau lagoon in relation with mortality events. Then, we compared mortality and growth parameters of oysters as a function of their origins, their development stages and cultivation sites in relation with energetic, reproduction and pathogens. Finally, we conducted transfer experiments where oysters maintained free of mortality in open sea were transferred in the Thau lagoon...
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Ano: 2010 URL: http://archimer.ifremer.fr/doc/00002/11354/7927.pdf
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