Differential reaction norms to ocean acidification in two oyster species from contrasting habitats.

Ocean acidification (OA), a consequence of the increase in anthropogenic emissions of carbon dioxide, causes major changes in the chemistry of carbonates in the ocean with deleterious effects on calcifying organisms. The pH/PCO2 range to which species are exposed in nature is important to consider when interpreting the response of coastal organisms to OA. In this context, emerging approaches, which assess the reaction norms of organisms to a wide pH gradient, are improving our understanding of tolerance thresholds and acclimation potential to OA. In this study, we deciphered the reaction norms of two oyster species living in contrasting habitats: the intertidal oyster Crassostrea gigas and the subtidal flat oyster Ostrea edulis, which are two economically and ecologically valuable species in temperate ecosystems. Six-month-old oysters of each species were exposed in common garden tanks for 48 days to a pH gradient ranging from 7.7 to 6.4 (total scale). Both species were tolerant down to a pH of 6.6 with high plasticity in fitness-related traits such as survival and growth. However, oysters underwent remodelling of membrane fatty acids to cope with decreasing pH along with shell bleaching impairing shell integrity and consequently animal fitness. Finally, our work revealed species-specific physiological responses and highlights that intertidal C. gigas seem to have a better acclimation potential to rapid and extreme OA changes than O. edulis. Overall, our study provides important data about the phenotypic plasticity and its limits in two oyster species, which is essential for assessing the challenges posed to marine organisms by OA.

An integrative perspective on fish health: Environmental and anthropogenic pathways affecting fish stress.

Multifactorial studies assessing the cumulative effects of natural and anthropogenic stressors on individual stress response are crucial to understand how organisms and populations cope with environmental change. We tested direct and indirect causal pathways through which environmental stressors affect the stress response of wild gilthead seabream in Mediterranean costal lagoons using an integrative PLS-PM approach. We integrated information on 10 environmental variables and 36 physiological variables into seven latent variables reflecting lagoons features and fish health. These variables concerned fish lipid reserves, somatic structure, inorganic contaminant loads, and individual trophic and stress response levels. This modelling approach allowed explaining 30 % of the variance within these 46 variables considered. More importantly, 54 % of fish stress response was explained by the dependent lagoon features, fish age, fish diet, fish reserve, fish structure and fish contaminant load latent variables included in our model. This integrative study sheds light on how individuals deal with contrasting environments and multiple ecological pressures.

Intertidal limits shape covariation between metabolic plasticity, oxidative stress and telomere dynamics in Pacific oyster (Crassostrea gigas).

In intertidal zones, species such as sessile shellfish exhibit extended phenotypic plasticity to face rapid environmental changes, but whether frequent exposure to intertidal limits of the distribution range impose physiological costs for the animal remains elusive. Here, we explored how phenotypic plasticity varied along foreshore range at multiple organization levels, from molecular to cellular and whole organism acclimatization, in the Pacific oyster (Crassostrea gigas). We exposed 7-month-old individuals for up to 16 months to three foreshore levels covering the vertical range for this species, representing 20, 50 and 80% of the time spent submerged monthly. Individuals at the upper range limit produced energy more efficiently, as seen by steeper metabolic reactive norms and unaltered ATP levels despite reduced mitochondrial density. By spending most of their time emerged, oysters mounted an antioxidant shielding concomitant with lower levels of pro-oxidant proteins and postponed age-related telomere attrition. Instead, individuals exposed at the lower limit range near subtidal conditions showed lower energy efficiencies, greater oxidative stress and shorter telomere length. These results unraveled the extended acclimatization strategies and the physiological costs of living too fast in subtidal conditions for an intertidal species.

Pacific oysters do not compensate growth retardation following extreme acidification events.

Ocean acidification caused by anthropogenic carbon dioxide emissions alters the growth of marine calcifiers. Although the immediate effects of acidification from global ocean models have been well studied on calcifiers, their recovery capacity over a wide range of pH has never been evaluated. This aspect is crucial because acidification events that arise in coastal areas can far exceed global ocean predictions. However, such acidification events could occur transiently, allowing for recovery periods during which the effects on growth would be compensated, maintained or amplified. Here we evaluated the recovery capacity of a model calcifier, the Pacific oyster Crassostrea gigas. We exposed juveniles to 15 pH conditions between 6.4 and 7.8 for 14 days. Oyster growth was retarded below pH 7.1 while shells were corroded at pH 6.5. We then placed the oysters under ambient pH > 7.8 for 42 days. Growth retardation persisted at pH levels below pH 7.1 even after the stress was removed. However, despite persistent retardation, growth has resumed rapidly suggesting that the oysters can recover from extreme acidification. Yet we found that the differences in individual weight between pH conditions below 7.1 increased over time, and thus the growth retardation cannot be compensated and may affect the fitness of the bivalves.

Microbiota of the Digestive Glands and Extrapallial Fluids of Clams Evolve Differently Over Time Depending on the Intertidal Position.

The Manila clam (Ruditapes philippinarum) is the second most exploited bivalve in the world but remains threatened by diseases and global changes. Their associated microbiota play a key role in their fitness and acclimation capacities. This study aimed at better understanding the behavior of clam digestive glands and extrapallial fluids microbiota at small, but contrasting spatial and temporal scales. Results showed that environmental variations impacted clam microbiota differently according to the considered tissue. Each clam tissue presented its own microbiota and showed different dynamics according to the intertidal position and sampling period. Extrapallial fluids microbiota was modified more rapidly than digestive glands microbiota, for clams placed on the upper and lower intertidal position, respectively. Clam tissues could be considered as different microhabitats for bacteria as they presented different responses to small-scale temporal and spatial variabilities in natural conditions. These differences underlined a more stringent environmental filter capacity of the digestive glands.

Harsh intertidal environment enhances metabolism and immunity in oyster (Crassostrea gigas) spat.

The Pacific oyster Crassostrea gigas is established in the marine intertidal zone, experiencing rapid and highly dynamic environmental changes throughout the tidal cycle. Depending on the bathymetry, oysters face oxygen deprivation, lack of nutrients, and high changes in temperature during alternation of the cycles of emersion/immersion. Here we showed that intertidal oysters at a bathymetry level of 3 and 5 m delayed by ten days the onset of mortality associated with Pacific Oyster Mortality Syndrome (POMS) as compared to subtidal oysters. Intertidal oysters presented a lower growth but similar energetic reserves to subtidal oysters but induced proteomic changes indicative of a boost in metabolism, inflammation, and innate immunity that may have improved their resistance during infection with the Ostreid herpes virus. Our work highlights that intertidal harsh environmental conditions modify host-pathogen interaction and improve oyster health. This study opens new perspectives on oyster farming for mitigation strategies based on tidal height.

Gene expression plasticity, genetic variation and fatty acid remodelling in divergent populations of a tropical bivalve species.

Ocean warming challenges marine organisms' resilience, especially for species experiencing temperatures close to their upper thermal limits. A potential increase in thermal tolerance might significantly reduce the risk of population decline, which is intrinsically linked to variability in local habitat temperatures. Our goal was to assess the plastic and genetic potential of response to elevated temperatures in a tropical bivalve model, Pinctada margaritifera. We benefit from two ecotypes for which local environmental conditions are characterized by either large diurnal variations in the tide pools (Marquesas archipelago) or low mean temperature with stable to moderate seasonal variations (Gambier archipelago). We explored the physiological basis of individual responses to elevated temperature, genetic divergence as well as plasticity and acclimation by combining lipidomic and transcriptomic approaches. We show that P. margaritifera has certain capacities to adjust to long-term elevated temperatures that was thus far largely underestimated. Genetic variation across populations overlaps with gene expression and involves the mitochondrial respiration machinery, a central physiological process that contributes to species thermal sensitivity and their distribution ranges. Our results present evidence for acclimation potential in P. margaritifera and urge for longer term studies to assess populations resilience in the face of climate change.

Le réchauffement des océans remet en question la résilience des organismes marins, en particulier pour les espèces connaissant des températures proches de leurs limites thermiques supérieures. Une augmentation potentielle de la tolérance thermique pourrait ainsi réduire considérablement le risque de déclin de la population. L'objectif de cette étude était d'évaluer le potentiel plastique et génétique de la réponse à l'exposition courte et chronique à températures élevées chez une espèce de bivalve tropical, Pinctada margaritifera. Ce modèle bénéficie de l'existence de deux écotypes pour lesquels les conditions environnementales locales sont caractérisées soit par de fortes variations diurnes associées aux marées (archipel des Marquises) soit par une température moyenne plus basse et des variations saisonnières prononcées (archipel des Gambier). Nous avons exploré les bases physiologiques des réponses individuelles ainsi que la divergence génétique et quantifié la plasticité en combinant des approches lipidomique et transcriptomique. Nous montrons que P. margaritifera possède des capacités d'acclimatation à des températures élevées sur le long terme jusqu'à présent largement sous-estimées. La divergence génétique entre populations est par ailleurs associée à des différences d'expression des gènes et implique la machinerie respiratoire mitochondriale, un processus physiologique central qui contribue à la sensibilité thermique des espèces et à leurs répartitions. Nos résultats présentent les bases des potentiels d'acclimatation chez P. margaritifera et soulignent l'importance d'études à plus long terme pour évaluer la résilience des populations face au changement climatique.

Seaweeds influence oyster microbiota and disease susceptibility.

A growing awareness of role that microbiota can play in mediating the effects of pathogens on hosts has given rise to the concept of the pathobiome. Recently, we demonstrated that the Pacific oyster mortality syndrome affecting Crassostrea gigas oysters is caused by infection with the Ostreid herpesvirus type 1 (OsHV-1) followed by infection with multiple bacterial taxa. Here we extend the concept of this pathobiome beyond the host species and its bacterial microbiota by investigating how seaweed living in association with oysters influences their response to the disease. We hypothesized that by their mere presence in the environment, different species of seaweeds can positively or negatively influence the risk of disease in oysters by shaping their bacterial microbiota and their immune response. Although seaweed and oysters do not have direct ecological interactions, they are connected by seawater and likely share microbes. To test our hypothesis, oysters were acclimated with green, brown or red algae for 2 weeks and then challenged with OsHV-1. We monitored host survival and pathogen proliferation and performed bacterial microbiota and transcriptome analyses. We found that seaweeds can alter the bacterial microbiota of the host and its response to the disease. More particularly, green algae belonging to the genus Ulva spp. induced bacterial microbiota dysbiosis in oyster and modification of its transcriptional immune response leading to increased susceptibility to the disease. This work provides a better understanding of a marine disease and highlights the importance of considering both macrobiotic and microbiotic interactions for conservation, management and exploitation of marine ecosystems and resources.

Revisiting tolerance to ocean acidification: Insights from a new framework combining physiological and molecular tipping points of Pacific oyster.

Studies on the impact of ocean acidification on marine organisms involve exposing organisms to future acidification scenarios, which has limited relevance for coastal calcifiers living in a mosaic of habitats. Identification of tipping points beyond which detrimental effects are observed is a widely generalizable proxy of acidification susceptibility at the population level. This approach is limited to a handful of studies that focus on only a few macro-physiological traits, thus overlooking the whole organism response. Here we develop a framework to analyze the broad macro-physiological and molecular responses over a wide pH range in juvenile oyster. We identify low tipping points for physiological traits at pH 7.3-6.9 that coincide with a major reshuffling in membrane lipids and transcriptome. In contrast, a drop in pH affects shell parameters above tipping points, likely impacting animal fitness. These findings were made possible by the development of an innovative methodology to synthesize and identify the main patterns of variations in large -omic data sets, fitting them to pH and identifying molecular tipping points. We propose the broad application of our framework to the assessment of effects of global change on other organisms.

Environmental optima for an ecosystem engineer: a multidisciplinary trait-based approach.

A complex interplay of biotic and abiotic factors underpins the distribution of species and operates across different levels of biological organization and life history stages. Understanding ecosystem engineer reproductive traits is critical for comprehending and managing the biodiversity-rich habitats they create. Little is known about how the reproduction of the reef-forming worm, Sabellaria alveolata, varies across environmental gradients. By integrating broad-scale environmental data with in-situ physiological data in the form of biochemical traits, we identified and ranked the drivers of intraspecific reproductive trait variability (ITV). ITV was highest in locations with variable environmental conditions, subjected to fluctuating temperature and hydrodynamic conditions. Our trait selection pointed to poleward sites being the most physiologically stressful, with low numbers of irregularly shaped eggs suggesting potentially reduced reproductive success. Centre-range individuals allocated the most energy to reproduction, with the highest number of intermediate-sized eggs, whilst equatorward sites were the least physiologically stressful, thus confirming the warm-adapted nature of our model organism. Variation in total egg diameter and relative fecundity were influenced by a combination of environmental conditions, which changed depending on the trait and sampling period. An integrated approach involving biochemical and reproductive traits is essential for understanding macro-scale patterns in the face of anthropogenic-induced climate change across environmental and latitudinal gradients.

The Pacific Oyster Mortality Syndrome, a Polymicrobial and Multifactorial Disease: State of Knowledge and Future Directions.

The Pacific oyster (Crassostreae gigas) has been introduced from Asia to numerous countries around the world during the 20th century. C. gigas is the main oyster species farmed worldwide and represents more than 98% of oyster production. The severity of disease outbreaks that affect C. gigas, which primarily impact juvenile oysters, has increased dramatically since 2008. The most prevalent disease, Pacific oyster mortality syndrome (POMS), has become panzootic and represents a threat to the oyster industry. Recently, major steps towards understanding POMS have been achieved through integrative molecular approaches. These studies demonstrated that infection by Ostreid herpesvirus type 1 µVar (OsHV-1 µvar) is the first critical step in the infectious process and leads to an immunocompromised state by altering hemocyte physiology. This is followed by dysbiosis of the microbiota, which leads to a secondary colonization by opportunistic bacterial pathogens, which in turn results in oyster death. Host and environmental factors (e.g. oyster genetics and age, temperature, food availability, and microbiota) have been shown to influence POMS permissiveness. However, we still do not understand the mechanisms by which these different factors control disease expression. The present review discusses current knowledge of this polymicrobial and multifactorial disease process and explores the research avenues that must be investigated to fully elucidate the complexity of POMS. These discoveries will help in decision-making and will facilitate the development of tools and applied innovations for the sustainable and integrated management of oyster aquaculture.

High temperature induces transcriptomic changes in Crassostrea gigas that hinder progress of ostreid herpesvirus (OsHV-1) and promote survival.

Of all environmental factors, seawater temperature plays a decisive role in triggering marine diseases. Like fever in vertebrates, high seawater temperature could modulate the host response to pathogens in ectothermic animals. In France, massive mortality of Pacific oysters, Crassostrea gigas, caused by the ostreid herpesvirus 1 (OsHV-1) is markedly reduced when temperatures exceed 24°C in the field. In the present study we assess how high temperature influences the host response to the pathogen by comparing transcriptomes (RNA sequencing) during the course of experimental infection at 21°C (reference) and 29°C. We show that high temperature induced host physiological processes that are unfavorable to the viral infection. Temperature influenced the expression of transcripts related to the immune process and increased the transcription of genes related to the apoptotic process, synaptic signaling and protein processes at 29°C. Concomitantly, the expression of genes associated with catabolism, metabolite transport, macromolecule synthesis and cell growth remained low from the first stage of infection at 29°C. Moreover, viral entry into the host might have been limited at 29°C by changes in extracellular matrix composition and protein abundance. Overall, these results provide new insights into how environmental factors modulate host-pathogen interactions.

Latitudinal drivers of oyster mortality: deciphering host, pathogen and environmental risk factors.

Diseases pose an ongoing threat to aquaculture, fisheries and conservation of marine species, and determination of risk factors of disease is crucial for management. Our objective was to decipher the effects of host, pathogen and environmental factors on disease-induced mortality of Pacific oysters (Crassostrea gigas) across a latitudinal gradient. We deployed young and adult oysters at 13 sites in France and we monitored survival, pathogens and environmental parameters. The young oysters came from either the wild collection or the hatchery while the adults were from the wild only. We then used Cox regression models to investigate the effect of latitude, site, environmental factors and origin on mortality risk and to extrapolate this mortality risk to the distribution limits of the species in Europe. We found that seawater temperature, food level, sea level atmospheric pressure, rainfall and wind speed were associated with mortality risk. Their effect on hatchery oysters was generally higher than on wild animals, probably reflecting that hatchery oysters were free of Ostreid herpesvirus 1 (OsHV-1) whereas those from the wild were asymptomatic carriers. The risk factors involved in young and adult oyster mortalities were different, reflecting distinct diseases. Mortality risk increases from 0 to 90% with decreasing latitude for young hatchery oysters, but not for young wild oysters or adults. Mortality risk was higher in wild oysters than in hatchery ones at latitude > 47.6°N while this was the opposite at lower latitude. Therefore, latitudinal gradient alters disease-induced mortality risk but interacts with the initial health status of the host and the pathogen involved. Practically, we suggest that mortality can be mitigated by using hatchery oysters in north and wild collected oysters in the south.

The marine intertidal zone shapes oyster and clam digestive bacterial microbiota.

Digestive microbiota provide a wide range of beneficial effects on host physiology and are therefore likely to play a key role in marine intertidal bivalve ability to acclimatize to the intertidal zone. This study investigated the effect of intertidal levels on the digestive bacterial microbiota of oysters (Crassostrea gigas) and clams (Ruditapes philippinarum), two bivalves with different ecological niches. Based on 16S rRNA region sequencing, digestive glands, seawater and sediments harbored specific bacterial communities, dominated by operational taxonomic units assigned to the Mycoplasmatales,Desulfobacterales and Rhodobacterales orders, respectively. Field implantation modified digestive bacterial microbiota of both bivalve species according to their intertidal position. Rhodospirillales and Legionellales abundances increased in oysters and clams from the low intertidal level, respectively. After a 14-day depuration process, these effects were still observed, especially for clams, while digestive bacterial microbiota of oysters were subjected to more short-term environmental changes. Nevertheless, 3.5 months stay on an intertidal zone was enough to leave an environmental footprint on the digestive bacterial microbiota, suggesting the existence of autochthonous bivalve bacteria. When comparing clams from the three intertidal levels, 20% of the bacterial assemblage was shared among the levels and it was dominated by an operational taxonomic unit affiliated to the Mycoplasmataceae and Spirochaetaceae families.

High temperature induces transcriptomic changes in Crassostrea gigas that hinders progress of Ostreid herpesvirus (OsHV-1) and promotes survival.

Among all the environmental factors, seawater temperature plays a decisive role in triggering marine diseases. Like fever in vertebrates, high seawater temperature could modulate the host response to the pathogens in ectothermic animals. In France, massive mortality of Pacific oysters Crassostrea gigas caused by the ostreid herpesvirus 1 (OsHV-1) is markedly reduced when temperatures exceed 24°C in the field. In the present study we assess how high temperature influences the host response to the pathogen by comparing transcriptomes (RNA-sequencing) during the course of experimental infection at 21°C (reference) and 29°C. We show that high temperature induced host physiological processes that are unfavorable to the viral infection. Temperature influenced the expression of transcripts related to the immune process and increased the transcription of genes related to apoptotic process, synaptic signaling, and protein processes at 29°C. Concomitantly, the expression of genes associated to catabolism, metabolites transport, macromolecules synthesis and cell growth remained low since the first stage of infection at 29°C. Moreover, viral entry into the host might have been limited at 29°C by changes in extracellular matrix composition and protein abundance. Overall, these results provide new insights into how environmental factors modulate the host-pathogen interactions.

Deciphering the effect of food availability, growth and host condition on disease susceptibility in a marine invertebrate.

Food provisioning influences disease risk and outcome in animal populations in two ways. On the one hand, unrestricted food supply improves the physiological condition of the host and lowers its susceptibility to infectious disease, reflecting a trade-off between immunity and other fitness-related functions. On the other hand, food scarcity limits the resources available to the pathogen and slows the growth and metabolism of the host on which the pathogen depends to proliferate. Here, we investigated how food availability, growth rate and energetic reserves drive the outcome of a viral disease affecting an ecologically relevant model host, the Pacific oyster, Crassostrea gigas We selected fast- and slow-growing animals, and we exposed them to high and low food rations. We evaluated their energetic reserves, challenged them with a pathogenic virus, monitored daily survival and developed a mortality risk model. Although high food levels and oyster growth were associated with a higher risk of mortality, energy reserves were associated with a lower risk. Food availability acts both as an enabling factor for mortality by increasing oyster growth and as a limiting factor by increasing their energy reserves. This study clarifies how food resources have an impact on susceptibility to disease and indicates how the host's physiological condition could mitigate epidemics. Practically, we suggest that growth should be optimized rather than maximized, considering that trade-offs occur with disease resistance or tolerance.

Dynamics of the Pacific oyster pathobiota during mortality episodes in Europe assessed by 16S rRNA gene profiling and a new target enrichment next-generation sequencing strategy.

Infectious agents such as the bacteria Vibrio aestuarianus or Ostreid herpesvirus 1 have been repeatedly associated with dramatic disease outbreaks of Crassostrea gigas beds in Europe. Beside roles played by these pathogens, microbial infections in C. gigas may derive from the contribution of a larger number of microorganisms than previously thought, according to an emerging view supporting the polymicrobial nature of bivalve diseases. In this study, the microbial communities associated with a large number of C. gigas samples collected during recurrent mortality episodes at different European sites were investigated by real-time PCR and 16SrRNA gene-based microbial profiling. A new target enrichment next-generation sequencing protocol for selective capturing of 884 phylogenetic and virulence markers of the potential microbial pathogenic community in oyster tissue was developed allowing high taxonomic resolution analysis of the bivalve pathobiota. Comparative analysis of contrasting C. gigas samples conducted using these methods revealed that oyster experiencing mortality outbreaks displayed signs of microbiota disruption associated with the presence of previously undetected potential pathogenic microbial species mostly belonging to genus Vibrio and Arcobacter. The role of these species and their consortia should be targeted by future studies aiming to shed light on mechanisms underlying polymicrobial infections in C. gigas.

Fine-scale temporal dynamics of herpes virus and vibrios in seawater during a polymicrobial infection in the Pacific oyster Crassostrea gigas.

The Pacific oyster Crassostrea gigas is currently being impacted by a polymicrobial disease that involves early viral infection by ostreid herpesvirus-1 (OsHV-1) followed by a secondary bacterial infection leading to death. A widely used method of inducing infection consists of placing specific pathogen-free oysters ('recipients') in cohabitation in the laboratory with diseased oysters that were naturally infected in the field ('donors'). With this method, we evaluated the temporal dynamics of pathogen release in seawater and the cohabitation time necessary for disease transmission and expression. We showed that OsHV-1 and Vibrio spp. in the seawater peaked concomitantly during the first 48 h and decreased thereafter. We found that 1.5 h of cohabitation with donors was enough time to transmit pathogens to recipients and to induce mortality later, reflecting the highly contagious nature of the disease. Finally, mortality of recipients was associated with increasing cohabitation time with donors until reaching a plateau at 20%. This reflects the cumulative effect of exposure to pathogens. The optimal cohabitation time was 5-6 d, the mortality of recipients occurring 1-2 d earlier.

Determination of risk factors for herpesvirus outbreak in oysters using a broad-scale spatial epidemiology framework.

Marine diseases have major impacts on ecosystems and economic consequences for aquaculture and fisheries. Understanding origin, spread and risk factors of disease is crucial for management, but data in the ocean are limited compared to the terrestrial environment. Here we investigated how the marine environment drives the spread of viral disease outbreak affecting The Pacific oyster worldwide by using a spatial epidemiology framework. We collected environmental and oyster health data at 46 sites spread over an area of 300 km2 along an inshore-offshore gradient during an epizootic event and conducted risk analysis. We found that disease broke out in the intertidal farming area and spread seaward. Mortalities and virus detection were observed in oysters placed 2 km from the farming areas, but oysters of almost all sites were subclinically infected. Increasing food quantity and quality, growth rate and energy reserves of oyster were associated with a lower risk of mortality offshore whereas increasing turbidity, a proxy of the concentration of suspended particulate matter, and terrestrial inputs, inferred from fatty acid composition of oysters, were associated with a higher risk of mortality. Offshore farming and maintenance of good ecological status of coastal waters are options to limit disease risk in oysters.

Temperature modulate disease susceptibility of the Pacific oyster Crassostrea gigas and virulence of the Ostreid herpesvirus type 1.

Temperature triggers marine diseases by changing host susceptibility and pathogen virulence. Oyster mortalities associated with the Ostreid herpesvirus type 1 (OsHV-1) have occurred seasonally in Europe when the seawater temperature range reaches 16-24 °C. Here we assess how temperature modulates oyster susceptibility to OsHV-1 and pathogen virulence. Oysters were injected with OsHV-1 suspension incubated at 21 °C, 26 °C and 29 °C and were placed in cohabitation with healthy oysters (recipients) at these three temperatures according to a fractional factorial design. Survival was followed for 14 d and recipients were sampled for OsHV-1 DNA quantification and viral gene expression. The oysters were all subsequently placed at 21 °C to evaluate the potential for virus reactivation, before being transferred to oyster farms to evaluate their long-term susceptibility to the disease. Survival of recipients at 29 °C (86%) was higher than at 21 °C (52%) and 26 °C (43%). High temperature (29 °C) decreased the susceptibility of oysters to OsHV-1 without altering virus infectivity and virulence. At 26 °C, the virulence of OsHV-1 was enhanced. Differences in survival persisted when the recipients were all placed at 21 °C, suggesting that OsHV-1 did not reactivate. Additional oyster mortality followed the field transfer, but the overall survival of oysters infected at 29 °C remained higher.