Genetic effect of domestication selective pressures on Pacific oyster at larval stage

Among bivalve species, the Pacific oyster, Crassostrea gigas, is the most economically important bivalve production over the world. Today, C. gigas is subject to an important production effort that leads to an intensive artificial selection. Larval stage is relatively unknown, specifically in a domestication context. Genetic consequence of artificial selection is still at a preliminary study. We aimed to tackle the consequence of inconscient domestication on the variance reproductive success focusing on larval stage, keystone of the life cycle. We studied two kinds of specific selective processes that common hatchery rearing practices exert : the effect of discarding the smallest larvae on genetic diversity and the artificial environment rearing effect via the temperature providing a contrast resembling wild versus hatchery conditions (20 and 26°C). In order to monitor the effect of the selection of fast growing larvae by sieving, growth variability and genetic diversity in a larval population descended from a factorial breeding was studied. We used a mixed-family approach to reduce potentially confounding environmental biais. The retrospective assignment of individuals to family groups has been performed using a three microsatellite markers set. Two different rearing were carried out in parallel. For three (replicates) 50-l tanks, the smallest larvae were progressively discarded by selective sieving, whereas for the three others no selective sieving was performed. The intensity of selective sieving was adjusted so as to discard 50% of the larvae over the whole rearing period in a progressive manner. As soon as the larvae reached the pediveliger stage, ready to settle larvae were sampled for genetic analysis. Regarding the artificial environment rearing effect via the temperature, we used a similar mixed-family approach. The progeny from a factorial breeding design was divided as follows: three (replicates) 50-l tanks were dedicaced to a rearing at 26°C versus 20°C for three others 50-l tanks. The whole size variability was preserved for this experiment. Individual growth measurements for larvae genetically identified have been performed at days 22 and 30 after fertilization for both conditions. In a same way, we collected individual measurements for genotyped juvenile oysters (80 days after fertilization). At a phenotypic scale, relative survival and settlement success for larvae with sieving were higher. Sieving appears as a time-saving process associated with a better relative survival ratio. But in the same time, our results confirm that a significant genetic variability exist for early developmental traits in the Pacific oyster. This is congruent with the results already obtained that investigated genetic variability and genetic correlations in early life-history traits of Crassostrea gigas. Discarding around 50% of the smallest larvae can lead to significant selection at the larval stage.

Disruption of amylase genes by RNA interference affects reproduction in the Pacific oyster Crassostrea gigas

Feeding strategies and digestive capacities can have important implications for variation in energetic pathways associated with ecological and economically important traits, such as growth or reproduction in bivalve species. Here, we investigated the role of amylase in the digestive processes of Crassostrea gigas, using in vivo RNA interference. This approach also allowed us to investigate the relationship between energy intake by feeding and gametogenesis in oysters. Double-stranded (ds)RNA designed to target the two α-amylase genes A and B was injected in vivo into the visceral mass of oysters at two doses. These treatments caused significant reductions in mean mRNA levels of the amylase genes: −50.7% and −59% mRNA A, and −71.9% and −70.6% mRNA B in 15 and 75 µg dsRNA-injected oysters, respectively, relative to controls. Interestingly, reproductive knock-down phenotypes were observed for both sexes at 48 days post-injection, with a significant reduction of the gonad area (−22.5% relative to controls) and germ cell under-proliferation revealed by histology. In response to the higher dose of dsRNA, we also observed reductions in amylase activity (−53%) and absorption efficiency (−5%). Based on these data, dynamic energy budget modeling showed that the limitation of energy intake by feeding that was induced by injection of amylase dsRNA was insufficient to affect gonadic development at the level observed in the present study. This finding suggests that other driving mechanisms, such as endogenous hormonal modulation, might significantly change energy allocation to reproduction, and increase the maintenance rate in oysters in response to dsRNA injection.

Mass mortality in bivalves and the intricate case of the Pacific oyster, Crassostrea gigas

Massive mortality outbreaks in cultured bivalves have been reported worldwide and they have been associated with infection by a range of viral and bacterial pathogens. Due to their economic and social impact, these episodes constitute a particularly sensitive issue in Pacific oyster (Crassostrea gigas) production. Since 2008, mortality outbreaks affecting C. gigas have increased in terms of intensity and geographic distribution. Epidemiologic surveys have lead to the incrimination of pathogens, specifically OsHV-1 and bacteria of the Vibrio genus, in particular Vibrio aestuarianus. Pathogen diversity may partially account for the variability in the outcome of infections. Host factors (age, reproductive status…) including their genetic background that has an impact on host susceptibility towards infection, also play a role herein. Finally, environmental factors have significant effects on the pathogens themselves, on the host and on the host-pathogen interaction. Further knowledge on pathogen diversity, classification, and spread, may contribute towards a better understanding of this issue and potential ways to mitigate the impact of these outbreaks.

Energy and Antioxidant Responses of Pacific Oyster Exposed to Trace Levels of Pesticides

Here, we assess the physiological effects induced by environmental concentrations of pesticides in Pacific oyster Crassostrea gigas. Oysters were exposed for 14 d to trace levels of metconazole (0.2 and 2 mu g/L), isoproturon (0.1 and 1 mu g/L), or both in a mixture (0.2 and 0.1 mu g/L, respectively). Exposure to trace levels of pesticides had no effect on the filtration rate, growth, and energy reserves of oysters. However, oysters exposed to metconazole and isoproturon showed an overactivation of the sensing-kinase AMP-activated protein kinase alpha (AMPK alpha), a key enzyme involved in energy metabolism and more particularly glycolysis. In the meantime, these exposed oysters showed a decrease in hexokinase and pyruvate kinase activities, whereas 2-DE proteomic revealed that fructose-1,6-bisphosphatase (F-1,6-BP), a key enzyme of gluconeogenesis, was upregulated. Activities of antioxidant enzymes were higher in oysters exposed to the highest pesticide concentrations. Both pesticides enhanced the superoxide dismutase activity of oysters. Isoproturon enhanced catalase activity, and metconazole enhanced peroxiredoxin activity. Overall, our results show that environmental concentrations of metconazole or isoproturon induced subtle changes in the energy and antioxidant metabolisms of oysters.

Global change and climate-driven invasion of the Pacific oyster (Crassostrea gigas) along European coasts: a bioenergetics modelling approach

Aim The spread of non-indigenous species in marine ecosystems world-wide is one of today's most serious environmental concerns. Using mechanistic modelling, we investigated how global change relates to the invasion of European coasts by a non-native marine invertebrate, the Pacific oyster Crassostrea gigas. Location Bourgneuf Bay on the French Atlantic coast was considered as the northern boundary of C. gigas expansion at the time of its introduction to Europe in the 1970s. From this latitudinal reference, variations in the spatial distribution of the C. gigas reproductive niche were analysed along the north-western European coast from Gibraltar to Norway. Methods The effects of environmental variations on C. gigas physiology and phenology were studied using a bioenergetics model based on Dynamic Energy Budget theory. The model was forced with environmental time series including in situ phytoplankton data, and satellite data of sea surface temperature and suspended particulate matter concentration. Results Simulation outputs were successfully validated against in situ oyster growth data. In Bourgneuf Bay, the rise in seawater temperature and phytoplankton concentration has increased C. gigas reproductive effort and led to precocious spawning periods since the 1960s. At the European scale, seawater temperature increase caused a drastic northward shift (1400 km within 30 years) in the C. gigas reproductive niche and optimal thermal conditions for early life stage development. Main conclusions We demonstrated that the poleward expansion of the invasive species C. gigas is related to global warming and increase in phytoplankton abundance. The combination of mechanistic bioenergetics modelling with in situ and satellite environmental data is a valuable framework for ecosystem studies. It offers a generic approach to analyse historical geographical shifts and to predict the biogeographical changes expected to occur in a climate-changing world.

Parental exposure to the herbicide diuron results in oxidative DNA damage to germinal cells of the Pacific oyster Crassostrea gigas

Chemical pollution by pesticides has been identified as a possible contributing factor to the massive mortality outbreaks observed in Crassostrea gigas for several years. A previous study demonstrated the vertical transmission of DNA damage by subjecting oyster genitors to the herbicide diuron at environmental concentrations during gametogenesis. This trans-generational effect occurs through damage to genitor-exposed gametes, as measured by the comet-assay. The presence of DNA damage in gametes could be linked to the formation of DNA damage in other germ cells. In order to explore this question, the levels and cell distribution of the oxidized base lesion 8-oxodGuo were studied in the gonads of exposed genitors. High-performance liquid chromatography coupled with UV and electrochemical detection analysis showed an increase in 8-oxodGuo levels in both male and female gonads after exposure to diuron. Immunohistochemistry analysis showed the presence of 8-oxodGuo at all stages of male germ cells, from early to mature stages. Conversely, the oxidized base was only present in early germ cell stages in female gonads. These results indicate that male and female genitors underwent oxidative stress following exposure to diuron, resulting in DNA oxidation in both early germ cells and gametes, such as spermatozoa, which could explain the transmission of diuron-induced DNA damage to offspring. Furthermore, immunostaining of early germ cells seems indicates that damages caused by exposure to diuron on germ line not only affect the current sexual cycle but also could affect future gametogenesis.

Involvement of Mitochondrial Activity and OXPHOS in ATP Synthesis During the Motility Phase of Spermatozoa in the Pacific Oyster, Crassostrea gigas

In the Pacific oyster, spermatozoa are characterized by a remarkably long movement phase (i.e., over 24 h) sustained by a capacity to maintain intracellular ATP level. To gain information on oxidative phosphorylation (OXPHOS) functionality during the motility phase of Pacific oyster spermatozoa, we studied 1) changes in spermatozoal mitochondrial activity, that is, mitochondrial membrane potential (MMP), and intracellular ATP content in relation to motion parameters and 2) the involvement of OXPHOS for spermatozoal movement using carbonyl cyanide m-chlorophenyl hydrazone (CCCP). The percentage of motile spermatozoa decreased over a 24 h movement period. MMP increased steadily during the first 9 h of the movement phase and was subsequently maintained at a constant level. Conversely, spermatozoal ATP content decreased steadily during the first 9 h postactivation and was maintained at this level during the following hours of the movement phase. When OXPHOS was decoupled by CCCP, the movement of spermatozoa was maintained 2 h and totally stopped after 4 h of incubation, whereas spermatozoa were still motile in the control after 4 h. Our results suggest that the ATP sustaining flagellar movement of spermatozoa may partially originate from glycolysis or from mobilization of stored ATP or from potential phosphagens during the first 2 h of movement as deduced by the decoupling by CCCP of OXPHOS. However, OXPHOS is required to sustain the long motility phase of Pacific oyster spermatozoa. In addition, spermatozoa may hydrolyze intracellular ATP content during the early part of the movement phase, stimulating mitochondrial activity. This stimulation seems to be involved in sustaining a high ATP level until the end of the motility phase.

The Kinome of Pacific Oyster Crassostrea gigas, Its Expression during Development and in Response to Environmental Factors

Oysters play an important role in estuarine and coastal marine habitats, where the majority of humans live. In these ecosystems, environmental degradation is substantial, and oysters must cope with highly dynamic and stressful environmental constraints during their lives in the intertidal zone. The availability of the genome sequence of the Pacific oyster Crassostrea gigas represents a unique opportunity for a comprehensive assessment of the signal transduction pathways that the species has developed to deal with this unique habitat. We performed an in silico analysis to identify, annotate and classify protein kinases in C. gigas, according to their kinase domain taxonomy classification, and compared with kinome already described in other animal species. The C. gigas kinome consists of 371 protein kinases, making it closely related to the sea urchin kinome, which has 353 protein kinases. The absence of gene redundancy in some groups of the C. gigas kinome may simplify functional studies of protein kinases. Through data mining of transcriptomes in C. gigas, we identified part of the kinome which may be central during development and may play a role in response to various environmental factors. Overall, this work contributes to a better understanding of key sensing pathways that may be central for adaptation to a highly dynamic marine environment.

OsHV-1 countermeasures to the Pacific oyster’s anti-viral response

The host-pathogen interactions between the Pacific oyster (Crassostrea gigas) and Ostreid herpesvirus type 1 (OsHV-1) are poorly characterised. Herpesviruses are a group of large, DNA viruses that are known to encode gene products that subvert their host’s antiviral response. It is likely that OsHV-1 has also evolved similar strategies as its genome encodes genes with high homology to C. gigas inhibitors of apoptosis (IAPs) and an interferon-stimulated gene (termed CH25H). The first objective of this study was to simultaneously investigate the expression of C. gigas and OsHV-1 genes that share high sequence homology during an acute infection. Comparison of apoptosis-related genes revealed that components of the extrinsic apoptosis pathway (TNF) were induced in response to OsHV-1 infection, but we failed to observe evidence of apoptosis using a combination of biochemical and molecular assays. IAPs encoded by OsHV-1 were highly expressed during the acute stage of infection and may explain why we didn’t observe evidence of apoptosis. However, C. gigas must have an alternative mechanism to apoptosis for clearing OsHV-1 from infected gill cells as we observed a reduction in viral DNA between 27 and 54 h post-infection. The reduction of viral DNA in C. gigas gill cells occurred after the up-regulation of interferon-stimulated genes (viperin, PKR, ADAR). In a second objective, we manipulated the host’s anti-viral response by injecting C. gigas with a small dose of poly I:C at the time of OsHV-1 infection. This small dose of poly I:C was unable to induce transcription of known antiviral effectors (ISGs), but these oysters were still capable of inhibiting OsHV-1 replication. This result suggests dsRNA induces an anti-viral response that is additional to the IFN-like pathway.

Ecophysiologie de la maturation sexuelle et de la ponte de l'huître creuse Crassostrea gigas : réponses métaboliques (respiration) et alimentaires (filtration, absorption) en fonction des différents stades de maturation

The study of sexual maturation and spawning in the Pacific oyster (Crassostrea gigas) is part of a vast research programme that endeavours to understand the causes of mortality that occur sporadically during the spring and summer seasons in the Marennes-Oléron Bay. Thermal and diet conditioning were used to obtain oysters at each stage of maturity simultaneously. Using the measured rates of clearance, consumption, absorption and respiration provided estimates of growth potential and gave the energetic budget of oysters at different stages of sexual maturity. Physiological responses were similar for males and females. Filtration decreased from 2.4 to 2.6 l.h (-1) to 1.8 l.h (-1) with increasing maturity. Weight gain was associated with gonad development and did not appear to have an effect on the clearance rate. Oysters 2.5 years old showed a negative energy budget (-15 J h (-1)) at later maturity stages. This deficit was confirmed (90 J.h (-1)) in oysters 1.5 years old at the same stage of maturity. On the contrary, immature oysters, in the early stages of maturity or post-spawning, had a growth potential of 110 to 170 J.h (-1). The energy deficit observed at later stages of maturity was primarily due to absorption, which decreased sharply during peak gametogenesis. Using measured respiration rates, an allometric relationship specific to gonad growth was determined with a coefficient of 0.574. Low physiological performance of oysters, observed at later stages of sexual maturity, must be taken into account in research on the factors responsible for spring and summer mortalities affecting oyster farms in Marennes-Oléron.

Apport d'un programme de génétique à une filière de production aquacole : l'exemple de l'ostréiculture

Two oyster species are currently present along the French coasts : the indigenous European flat oyster (Ostrea edulis), and the Pacific cupped oyster (Crassostrea gigas), that has been introduced from Japan since the beginning of the 70ies. The flat oyster successively suffered from two protozoan diseases during the 60ies and its production decreased from 20 000 tons/year by that time to 1 500 tons/year nowadays. Consequently, the oyster production is principally (99%) based upon the Pacific oyster species with approximately 150 000 tons/year among which 90% are grown from the natural spat. However, the hatchery production of this species is developing and was estimated to 400 to 800 millions spat in 2002. Moreover, strengthened relationships between IFREMER and the 5 commercial hatcheries, that all joined the SYSAAF (Union of the French poultry, shellfish and fish farming selectors), allow to plan for new genetic breeding programs. At the end of the 80ies, IFREMER initiated a genetic breeding program for the resistance of the European flat oyster to the bonamiosis, and obtained strains more tolerant to this disease. After two generations of massal selection, molecular markers had identified a reduced genetic basis in this program. It was then reoriented to an intra-familial selection. However, we were confronted to a zootechnic problem to manage such a scheme and we compromised by an intra-cohorts of families selection scheme managed using molecular markers. The program has now reached the transfer level with experimentation at a professional scale. Concerning the Pacific cupped oyster, and in parallel with the obtaining and the study of polyploids, performance of different Asian cupped oyster strains were compared to the one introduced in France thirty years ago and currently suffering from summer mortalities. The local strain exhibited better performance, certainly based upon a good local adaptation. In other respects, although early growth is a relevant criteria for selection for growth to commercial stage, it is not to be privileged in the context of an oyster producing region with a limited food availability. Contrary, the spat summer mortality became a priority for numerous teams (genetic, physiology, pathology, ecology,...) joined in the MOREST program. The first results showed important survival differences between fullsib and halsib families. They indicate a genetic determinism to this character "survival" and promote for its selection.

Relations entre apports terrigènes et conchyliculture dans les Pertuis Charentais

This work presents interactions between quantitative and qualitative river freshwater inputs and the shellfish farming (oyster and mussel) in the Pertuis Charentais. The quantity of freshwater (i.e. salinity) seems to have a weak influence on the shellfish farming contrarily to its quality determined by particulate and dissolved matters contained in the water. In autumn and winter, large precipitations have a "globally positive" effect amending the coastal ecosystem. Associated dissolved nutriments and the organic matter largely determine the quality of the coming spring growth for bred shellfish, itself controlling in turn the annual yield efficiencies. However, in winter their effects are postponed because of strong mineral load, low luminosity and temperature, then limiting the primary production. The spring contributions, directly linked to territorial practices, agriculture and tourism are more variable in quantity and quality from one year to another. They often correspond to high-risk inflows since numerous substances from anthropogenic watersheds can be found diluted in the coastal zone as in the Pertuis Charentais. Their impacts on in situ estuarine ecosystems are still poorly known since these substances are mainly studied and estimated in laboratory in controlled conditions. Several studies showed anthropogenic contaminations (i.e. cadmium, pesticides) could have significant direct or indirect effects on shellfish farming. For instance, the "summer" mortalities between 1990 and 2000 in the South of the Marennes-Oléron bay (MOB), that induced environmental and physiological oyster disorders, could be linked to pesticide effects, measured during consecutive years on the oyster bed of Ronce Perquis in the South of the MOB. The weak results from the spring larval rearing of the IFREMER experimental hatchery in the South of the bay, and chromosomal abnormalities measured on the stocks of wild oysters of the Pertuis could confirm a high-risk spring environment for the shellfish farming. In summer terrestrial inputs are reduced by low precipitations, anthropogenic water removals (drinking water, irrigation) and by plant evapotranspiration. Consequently certain years, a significant salinity increase in water masses of the Pertuis Charentais is observed. However, based on long-term observations, the significant interannual variability noticed in freshwater contributions constitutes one of the most important facts of these last years. When contributions are weak (i.e. 1991 and 2011), the mean annual salinity is 34.5 in the MOB. To the contrary, other years (i.e. 1977, 1981, 1983 and 1988), the mean salinity reduced to 30.5 shows the significant freshwater contributions to the bay. Elsewhere, particularly in the mediterranean region, oyster breeding water conditions characterized by high salinity values show the freshwater does not seem to be necessary for biological functions of the Pacific oyster Crassostrea gigas. Indeed, the oyster embryonic life in particular is well adapted to high salinity values as long as trophic resources are substantial and temperatures remain high. These two factors firstly condition the embryonic survival before the water salinity. Besides, in the Pertuis Charentais, wind conditions and the geographical bloodstock position rather determine the success of the larvae capture than seawater physic-chemical conditions. Finally, a misunderstanding still remains on summer freshwater contributions to the oyster larvae food supply.

Observer, Analyser et Gérer la variabilité de la reproduction et du recrutement de l’huître creuse en France : Le Réseau Velyger. Rapport annuel 2015

La conchyliculture, et principalement l’élevage de l’huître creuse, Crassostrea gigas, constitue la principale activité aquacole française. Cette activité repose, en grande partie, sur le recrutement naturel de l’espèce qui assure 60 à 70% des besoins en jeunes huîtres (naissain) : cette activité de collecte s’appelle le captage. Les deux principaux centres de captage en France sont les bassins d’Arcachon et de Marennes-Oléron. Or, depuis une dizaine d'années, sur le bassin d'Arcachon, le captage devient très variable: à des années de captage nul (par exemple 2002, 2005, 2007) ou faible (2009, 2010, 2011) succèdent des années excellentes voire pléthoriques (2003, 2006, 2008, 2012, 2014). A Marennes-Oléron, cette variabilité existe, mais s’avère beaucoup moins marquée. En outre, à la faveur du lent réchauffement des eaux, le captage peut désormais se pratiquer de plus en plus vers le nord. Ainsi, la baie de Bourgneuf, mais aussi la rade de Brest sont devenues, depuis quelques années, des secteurs où un nombre croissant d’ostréiculteurs pratiquent le captage avec succès, mais avec, là aussi, des irrégularités dans le recrutement qu’il convient de comprendre. Enfin, depuis la crise des mortalités de 2008, il se développe aussi sur la lagune de Thau une volonté de pratiquer le captage. Afin de mieux comprendre les facteurs de variations du captage, l’Ifremer a mis en place, à la demande du Comité National de la Conchyliculture, un réseau national de suivi de la reproduction : le Réseau Velyger. Créé en 2008 sur financements européens et financé désormais par la Direction des Pêches Maritimes et de l’Aquaculture, ce réseau apporte, chaque année, sur les écosystèmes cités précédemment, une série d’indicateurs biologiques (maturation, fécondité, date de ponte, abondance et survie larvaire, intensité du recrutement, survie du naissain) dont l’analyse croisée avec des indicateurs hydrologiques et climatiques permet progressivement de mieux appréhender les causes de variabilité du recrutement de l’huître creuse en France, modèle biologique et espèce clé de la conchyliculture française. Ce rapport présente donc les résultats 2015 de ce réseau d’observation et fait appel, pour la partie hydro-climatique, à des données acquises par d’autres réseaux régionaux et nationaux. Il détaille et analyse par site toutes les caractéristiques du cycle de reproduction de l’huître creuse : maturation et fécondité des adultes, période de ponte, abondance et survie des larves, intensité du captage et mortalités précoces. Il fournit ensuite une interprétation et une synthèse des résultats 2015 à la lueur des résultats des années antérieures. Ainsi, pour l’année 2015, on retient les faits majeurs suivants : • Sur le plan hydro-climatique, cette année se caractérise par un hiver doux et un printemps dans les normales, suivis d’un été là aussi très proches des normales à quelques exceptions près : l’étang de Thau affiche tout au long de l’été des températures largement excédentaires. Compte tenu d’une pluviométrie là aussi proche des normales, les concentrations en phytoplancton sont restées à un niveau moyen de la rade de Brest aux pertuis charentais et plutôt déficitaires dans le bassin d’Arcachon et la lagune de Thau. • En termes de biologie, ces conditions hydro-climatiques se sont traduites, chez les populations d’huîtres adultes, par des indices de condition, proxy de la fécondité, généralement proches des moyennes, avec toujours l’existence d’un gradient nord-sud observé chaque année, corrélativement à la concentration en phytoplancton. En outre, l’absence d’excédent thermique au printemps et en début d’été n’a pas permis de ponte précoce (à l’exception de la lagune de Thau), elle a même été plutôt tardive surtout dans le bassin d’Arcachon. • Sur la façade atlantique, les températures de l’eau lors du développement larvaire des principales cohortes ont été plutôt basses (inférieures à 20°C en rade de Brest et inférieures à 21°C ailleurs) et donc la vitesse de croissance larvaire a été ralentie et la survie amoindrie. Les rendements larvaires ont été effectivement très bas (e.g. 0,002 % à Arcachon). In fine, il y a eu peu de larves grosses dans l’eau, ce qui s’est traduit par un captage faible à modéré. Une exception tout de même : dans la lagune de Thau, les températures caniculaires tout au long de l’été ont permis une concentration moyenne de larves ‘grosses’ modérée (80 larves/1,5m3). Cependant, les méthodes et les techniques de captage sont encore en cours d’optimisation sur ce secteur et cette année, malgré cette présence de larves grosses, le captage est resté faible (< 10 naissains par coupelle à l’automne). • En conséquence, l’année 2015, se caractérise par un captage globalement « faible à modéré » dans tous les secteurs s’échelonnant autour de 10 naissains/coupelle dans la lagune de Thau et en baie de Bourgneuf à plus de 200 naissains/coupelle dans les pertuis charentais. Enfin, à partir de l’ensemble des résultats acquis depuis 2008, ce rapport fournit en conclusion une série de recommandations à prendre en compte pour préserver le captage dans les années à venir.

Qualification zoo-sanitaire du naissain de captage 2013 de l'huître creuse Crassostrea gigas (QUALIF) - Détection précoce des maladies en épreuve thermique de laboratoire

Depuis 2008, des mortalités massives d’huîtres creuses âgées de moins d’un an sont relevées sur le littoral français dès que la température de l’eau de mer atteint le seuil de 16°C. Ces mortalités de naissains sont associées à la détection du virus OsHV-1 μVar. Des travaux de qualification zoo-sanitaire menés depuis 2010 ont montré qu’un lot de naissains issus du captage naturel sur deux était infecté par le virus bien avant l’apparition des mortalités dans le milieu naturel, mais qu’a contrario les naissains d’écloserie expertisés présentaient une fréquence plus faible de portage latent. Dans ce contexte de crise zoo-sanitaire chronique, l’objectif de notre étude consistait à obtenir une information du statut zoo-sanitaire OsHV-1 μVar des populations de naissains du captage 2013 à l’échelle national. Ce travail avait pour finalité de permettre l’identification précoce des zones de captage associées à un risque de mortalité due à l’infection par OsHV-1 μVar. La méthodologie de l’épreuve thermique de laboratoire (ETL) correspond en une période de 1 mois d’isolement des naissains à évaluer en conditions contrôlées de laboratoire. Lors de cette période la température de l’eau de mer est maintenue constante à 21°C et la mortalité est relevée tous les 10 jours. La survie finale associée à des analyses qPCR permettent de qualifier le statut sanitaire en terme de portage OsHV-1 μVar de l’échantillon (statut infecté ou non infecté). Entre février et mars 2014, vingt et un échantillons de naissains âgés de 5 à 7 mois ont été échantillonnés dans six sites du littoral français (étang de Thau, bassin d’Arcachon, pertuis Charentais, baie de Bourgneuf, baie de Vilaine et rade de Brest) pour être qualifiés par ETL au site expérimental Ifremer d’Argenton. Les résultats de la campagne 2014 de qualification zoo-sanitaire confirment qu’en période hivernale, dans le milieu naturel, des naissains de captage peuvent être infectés par OsHV-1 μVar sans développement apparent de maladies ni mortalité. En revanche en ETL, ces lots de naissains infectés par OsHV-1 μVar présentent des taux de mortalité importants. Ainsi, sur les 20 échantillons de naissains de captage étudiés, onze ont révélé des maladies en ETL avec des mortalités cumulées variant de 16 % à 80 %. Ce ratio d’un échantillon sur deux de naissains infectés par OsHV-1 μVar demeure proche de celui observé de 2010 à 2013 lors des travaux précédents qui avaient permis de définir l’ETL. les six sites étudiés ont montré des résultats contrastés. En effet, les résultats se sont révélés favorables pour l’étang de Thau et la baie de Bourgneuf et défavorables dans le cas de la rade de Brest et la Baie de Vilaine. Pour ces deux derniers sites, tous les échantillons de naissains testés ont montré de fortes mortalités pendant l’ETL. Des différences s’observent également intra-site, notamment à Marennes Oléron avec 1 échantillon sur 3 infectés par OsHV-1 μVar ou dans pour Arcachon avec 3 échantillons sur 5. Dans le contexte préoccupant de l’épizootie actuelle, nos résultats confirment que certains lots de naissains de captage sont infectés par OsHV-1 μVar dès les mois de février-mars et pourrait être des acteurs actifs dans le déclenchement annuel du processus infectieux en milieu naturel quand la température de l’eau dépasse 16°C. Par ailleurs, notre étude démontre qu’il serait possible chaque année d’obtenir une qualification sanitaire précoce des zones de captage. Cela permettrait d’améliorer la gestion des risques liés aux transferts de naissains infectés par OsHV-1 μVar dans les différents bassins de production.

Qualification zoo-sanitaire du naissain de captage 2014 de l'huître creuse Crassostrea gigas (QUALIF) - Détection précoce des maladies en épreuve thermique de laboratoire

Depuis 2008, des mortalités massives d’huîtres creuses âgées de moins d’un an sont relevées sur le littoral français dès que la température de l’eau de mer atteint le seuil de 16°C. Ces mortalités de naissains sont associées à la détection du virus OsHV-1 μVar. Depuis 2010, des travaux de qualification zoo-sanitaire ont montré qu’environ un lot de naissains de captage sur deux était infecté par le virus avant l’apparition des mortalités dans le milieu naturel. En 2014, dans le cadre de la première action QUALIF, les résultats de la qualification zoo-sanitaire des naissains du captage de l’année 2013 à l’échelle nationale ont mis en évidence le caractère infectieux des maladies développées par ces naissains porteurs latents d’OsHV-1 μVar. L’objectif 2015 du second volet QUALIF consistait à reproduire le travail de qualification zoo-sanitaire OsHV-1 μVar pour les naissains du captage de l’année 2014. Les résultats (2014 et 2015) permettront d’aborder la variation interannuelle du statut zoo-sanitaire des naissains pour chacun des sites étudiés. La méthodologie de l’épreuve de qualification zoo-sanitaire consiste en une période de 1 mois d’isolement des naissains en conditions contrôlées de laboratoire. Lors de cette épreuve, la température de l’eau de mer est maintenue constante à 21°C et la mortalité est relevée tous les 10 jours. La survie finale et les analyses qPCR permettent de qualifier le statut sanitaire en terme de portage OsHV-1 μVar de l’échantillon (infecté ou non infecté). De début janvier à fin mars 2015, 39 échantillons de naissains âgés de 5 à 7 mois ont été prélevés dans 6 sites du littoral français (étang de Thau, bassin d’Arcachon, bassin de Marennes Oléron, baie de Bourgneuf, embouchure de La Vilaine et rade de Brest) pour être testés en épreuve thermique de laboratoire (ETL) dans l’outil expérimental Ifremer à Argenton. Les résultats montrent que 25 des 39 (soit 64 %) lots de naissains de captage testés ont développé des maladies en ETL associés à des mortalités cumulées variant de 6 à 72 %. Cette valeur de 64 % de lots détectés est plus élevée que celle précédemment observée en 2014 (55 %). Les résultats de cette seconde campagne de qualification zoo-sanitaire sont favorables pour les naissains de l’étang de Thau (absence de mortalité), moins favorables pour ceux d’Arcachon, Marennes Oléron, et ceux de la baie de Bourgneuf (réponses contrastées en terme de mortalité intra-site) et très défavorables pour les naissains de La Vilaine et de la rade de Brest (mortalités observées sur tous les échantillons testés). La principale conclusion de cette seconde étude de qualification zoo-sanitaire est identique à celle de 2014, à savoir qu’il est existe dans le milieu naturel en période hivernale des lots de naissains de captage infectés par OsHV-1 μVar sans développement apparent des maladies. Ces lots de naissains porteurs latents d’OsHV-1 μVar sont détectables en ETL et la présence d’OsHV-1 μVar peut alors être confirmée par qPCR. Dans le contexte actuel d’épizooties chroniques, nos résultats confirment à nouveau qu’il est possible d’identifier précocement les lots de naissains infectés ou non par OsHV-1 μVar. Ces animaux infectés sont un réservoir du virus en période hivernale. Dès que la température de l’eau de mer franchit le seuil de 16°C, ils participeront à la réémergence des maladies en milieu naturel. Par ailleurs, cette seconde étude de qualification zoo-sanitaire confirme la possibilité d’apprécier le risque sanitaire pour chaque zone de captage en fonction de la détection précoce des lots de naissains infectés par OsHV-1 μVar.