Ostreid Herpesvirus Research Articles

Potential of genomic selection for improvement of resistance to ostreid herpesvirus in Pacific oyster (Crassostrea gigas).

In genomic selection (GS), genome-wide SNP markers are used to generate genomic estimated breeding values for selection candidates. The application of GS in shellfish looks promising and has the potential to help in dealing with one of the main issues currently affecting Pacific oyster production worldwide, which is the 'summer mortality syndrome'. This causes periodic mass mortality in farms worldwide and has mainly been attributed to a specific variant of the ostreid herpesvirus (OsHV-1). In the current study, we evaluated the potential of genomic selection for host resistance to OsHV-1 in Pacific oysters, and compared it with pedigree-based approaches. An OsHV-1 disease challenge was performed using an immersion-based virus exposure treatment for oysters for 7days. A total of 768 samples were genotyped using the medium-density SNP array for oysters. A GWAS was performed for the survival trait using a GBLUP approach in blupf90 software. Heritability ranged from 0.25±0.05 to 0.37±0.05 (mean±SE) based on pedigree and genomic information respectively. Genomic prediction was more accurate than pedigree prediction, and SNP density reduction had little impact on prediction accuracy until marker densities dropped below approximately 500 SNPs. This demonstrates the potential for GS in Pacific oyster breeding programmes, and importantly, demonstrates that a low number of SNPs might suffice to obtain accurate genomic estimated breeding values, thus potentially making the implementation of GS more cost effective.

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

ABSTRACTOf 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.

Effect of air exposure, handling stress and imidacloprid on the susceptibility of Crassostrea gigas to Ostreid herpesvirus 1 (OsHV-1)

Effect of air exposure, handling stress and imidacloprid on the susceptibility of Crassostrea gigas to Ostreid herpesvirus 1 (OsHV-1)

First evaluation of resistance to both a California OsHV-1 variant and a French OsHV-1 microvariant in Pacific oysters

BackgroundVariants of the Ostreid herpesvirus 1 (OsHV-1) cause high losses of Pacific oysters globally, including in Tomales Bay, California, USA. A suite of new variants, the OsHV-1 microvariants (μvars), cause very high mortalities of Pacific oysters in major oyster-growing regions outside of the United States. There are currently no known Pacific oysters in the United States that are resistant to OsHV-1 as resistance has yet to be evaluated in these oysters. As part of an effort to begin genetic selection for resistance to OsHV-1, 71 families from the Molluscan Broodstock Program, a US West Coast Pacific oyster breeding program, were screened for survival after exposure to OsHV-1 in Tomales Bay. They were also tested in a quarantine laboratory in France where they were exposed to a French OsHV-1 microvariant using a plate assay, with survival recorded from three to seven days post-infection.ResultsSignificant heritability for survival were found for all time points in the plate assay and in the survival phenotype from a single mortality count in Tomales Bay. Genetic correlations between survival against the French OsHV-1 μvar in the plate assay and the Tomales Bay variant in the field trait were weak or non-significant.ConclusionsFuture breeding efforts will seek to validate the potential of genetic improvement for survival to OsHV-1 through selection using the Molluscan Broodstock Program oysters. The lack of a strong correlation in survival between OsHV-1 variants under this study’s exposure conditions may require independent selection pressure for survival to each variant in order to make simultaneous genetic gains in resistance.

Connectivities with shellfish farms and channel rivers are associated with mortality risk in oysters

AEI Aquaculture Environment Interactions Contact the journal Facebook Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsTheme Sections AEI 11:493-506 (2019) - DOI: https://doi.org/10.3354/aei00327 Connectivities with shellfish farms and channel rivers are associated with mortality risk in oysters Aline Gangnery1,*,**, Julien Normand1,**, Cyrielle Duval1, Philippe Cugier2, Karine Grangeré3, Bruno Petton4, Sébastien Petton4, Francis Orvain3, Fabrice Pernet5 1Ifremer, Laboratoire Environnement Ressources de Normandie, Avenue du Général de Gaulle, 14 520 Port en Bessin, France 2Ifremer, Laboratoire d’Ecologie Benthique Côtière, Technopole Brest-Iroise, 29 280 Plouzané, France 3Université de Caen Basse-Normandie, UMR BOREA, Esplanade de la Paix, BP 5186, 14 032 Caen cedex, France 4Ifremer, UMR 6539 LEMAR, 11 presqu’île du vivier, 29 840 Argenton en Landunvez, France 5Ifremer, UMR 6539 LEMAR, Technopole Brest-Iroise, 29 280 Plouzané, France *Corresponding author: aline.gangnery@ifremer.fr**These authors contributed equally to this work ABSTRACT: Oyster diseases have major consequences on fisheries and aquaculture. In France, young Pacific oysters Crassostrea gigas are severely hit by the ostreid herpesvirus, whereas adults suffer mortalities presumably caused by pathogenic bacteria. Here we investigated the origin and spread of mortalities that affect both young and adult oysters, and we identified and compared their risk factors. Mortality was monitored in 2 age classes of oysters deployed in early spring at 39 sites spread over a 37 km2 surface area inside and outside of shellfish farms. Environmental data obtained from numerical modelling were used to investigate risk factors. Mortality of young oysters associated with ostreid herpesvirus occurred in the oyster farming area. Hydrodynamic connectivity with oyster farms was associated with higher mortality risk, whereas chlorophyll a concentration was associated with a lower risk. Adult oysters experienced 2 mortality events that were associated with different risk factors. The first event, which occurred after deployment and was probably caused by endogenous pathogens, was mainly associated with connectivity to channel rivers and salinity. The second mortality event observed at the end of the summer was mainly associated with connectivity to oyster farms, suggesting pathogen transmission. The risk factors involved in young and adult oyster mortalities were partly different, reflecting distinct origins. Connectivity with oyster farms is a mortality risk factor for both young and adult oysters; thus, disease management strategies that focus on oyster farming areas will impact overall disease risk. KEY WORDS: Aquaculture · Bivalve · Disease · Epidemiology and health · Ecological modelling · Hydrodynamic connectivity Full text in pdf format Supplementary material PreviousNextCite this article as: Gangnery A, Normand J, Duval C, Cugier P and others (2019) Connectivities with shellfish farms and channel rivers are associated with mortality risk in oysters. Aquacult Environ Interact 11:493-506. https://doi.org/10.3354/aei00327 Export citation Mail this link - Contents Mailing Lists - RSS Facebook - Tweet - linkedIn Cited by Published in AEI Vol. 11. Online publication date: September 19, 2019 Print ISSN: 1869-215X; Online ISSN: 1869-7534 Copyright © 2019 Inter-Research.

Detection of isothermally amplified ostreid herpesvirus 1 DNA in Pacific oyster (Crassostrea gigas) using a miniaturised electrochemical biosensor

Given the threat that ostreid herpesvirus 1 (OsHV-1) poses to shellfish aquaculture, the need for rapid, user-friendly and cost-effective methods to detect this marine pathogen and minimise its impact is evident. In this work, an electrochemical biosensor for the detection of OsHV-1 based on isothermal recombinase polymerase amplification (RPA) was developed. The system was first tested and optimised on maleimide microtitre plates as a proof-of-concept, before being implemented on miniaturised gold electrodes. Amperometric detection of the isothermally amplified product was achieved through a sandwich hybridisation assay with an immobilised thiolated capture probe and a horseradish peroxidase (HRP)-labelled reporter probe. Calibration curves were constructed using PCR-amplified OsHV-1 DNA, achieving a limit of detection of 207 OsHV-1 target copies. The biosensor was applied to the analysis of 16 oyster samples from an infectivity experiment, and results were compared with those obtained by qPCR analysis, showing a strong degree of correlation (r = 0.988). The simplicity, rapidity, cost-effectiveness and potential for in-situ testing with the developed biosensor provide a valuable tool for the detection of OsHV-1 in aquaculture facilities, improving their management.

L'impact des microvariants du virus herpétique ostreid herpesvirus 1 sur la culture d'huîtres creuses dans les hémisphères Nord et Sud depuis 2008.

Mollusc farming is the third most productive aquaculture activity in the world, and the Pacific oyster (Crassostrea gigas) is one of the most important farmed species. Since 2008, mass mortalities in C. gigas due to ostreid herpesvirus 1 microvariants have challenged the viability of this industry in Europe, New Zealand and Australia. Ten years after the emergence of this disease, there is evidence that the industry has become consolidated into fewer, larger companies, with the displacement of small farming enterprises and loss of employment in coastal communities. Rather than seeking technical solutions, the industry has turned to compensatory production strategies, such as increasing the number of spat placed on farms, higher market prices for table oysters and direct marketing, which appear to have allowed profitability. Biosecurity policies and responses to outbreaks, including those from within the industry, have had unintended consequences for hatcheries and farmers in areas free of disease, mainly caused by restrictions on animal movements, and have not prevented global spread. There may be opportunities for better coordination of industry and government responses to epizootic disease emergence in aquaculture. There is certainly a need for increased adoption of technical advances from research, once these solutions have been adequately verified.

Long-term temporal and spatial patterns of Ostreid herpesvirus 1 (OsHV-1) infection and mortality in sentinel Pacific oyster spat (Crassostrea gigas) inform farm management

Disease due to Ostreid herpes virus − 1 (OsHV-1) is a major constraint to farming the Pacific oyster Crassotrea gigas in Europe and Australasia, requiring new management strategies. The objectives of this study were to determine the seasonal limits of transmission of OsHV-1, the spatial distribution of the virus, the seasonal water temperatures associated with OsHV-1 transmission and disease and the suitability of active surveillance for OsHV-1 using sentinel oysters. Spat were deployed and sampled regularly between 2012 and 2017 at 15 sites in two large estuaries and the times of OsHV-1 exposure and mortality were identified at weekly to monthly intervals. Mortality due to OsHV-1 was observed each summer season but was significantly more likely in the Georges than the Hawkesbury River estuary. Mortality was most widespread and frequent between December and April and was not observed prior to late October or after May. However, subclinical infections with OsHV-1 that did not progress to mortality were detected from early October until late June. There was a significant reduction in mortality due to OsHV-1 as well as subclinical infection with OsHV-1 between 2012–13 and 2016–17, and viral loads in infected oysters also appeared to decline after 2012–13. Mortality due to OsHV-1 was observed in sentinel spat at 14 of 15 sites but not at every site in every season. Different outcomes were sometimes seen between spat in paired baskets that were 1 to 100 m apart at the same site, attributable to differential exposure to OsHV-1 (clustering). Fifteen different batches of spat were used and mortality due to OsHV-1 occurred in all but three; both triploid and diploid spat were affected. In 2016–17, there was a trend of better survival in selected spat with potential genetic resistance to OsHV-1 compared to unselected spat. There were no biologically significant differences in the growth of spat that subsequently died due to OsHV-1 infection and those that survived. Mortality due to OsHV-1 commenced when the mean water temperature rose above approximately 20 °C in spring, which is 4–5 °C warmer than the initiation threshold in Europe. These consistent epidemiological patterns of infection and mortality will enable strategic placement of susceptible oysters in these estuaries at certain locations and within specific time windows with predicted low risk of mortality due to OsHV-1; water temperature monitoring may be used to predict this risk. Use of sentinel oysters for surveillance for OsHV-1 can be effective, but the clustering of exposure at all scales renders this more reliable at estuary level rather than at bay or site level and it requires an intensive sampling program. Non-specific mortality of spat was relatively common, therefore tests for OsHV-1 should be applied in field trials to distinguish mortality due to OsHV-1 from other causes of mortality.

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.

SummaryInfectious 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.

First detection of Ostreid herpesvirus 1 in wild Crassostrea gigas in Argentina

Ostreid herpesvirus 1 (OsHV-1) is a DNA virus of the genus Ostreavirus (Malacoherpesviridae family, Herpesvirales order). Worldwide, OsHV-1 and its microvariants have been associated with increased mortality of Pacific oysters, Crassostrea gigas. Adult asymptomatic oysters also have shown a high prevalence of viral infection. As a consequence, surveillance is needed to better describe OsHV-1 diversity, pathogenicity, clinical signs, and geographical distribution. We examined Crassostrea gigas sampled in October 2017 from the inner zone of the Bahía Blanca Estuary, Argentina, and found that 8 of 30 specimens (26.7%) presented macroscopic lesions in mantle tissues. Histological analysis revealed abnormal presentation of mantle epithelial cells and connective tissues. Conventional and real-time PCR conducted on the oyster samples revealed 70% to be positive for presence of OsHV-1 DNA. The nucleotide sequence of the amplicon obtained from one sample using the primer pair IA1/IA2 (targeting ORF 42/43) was 99% identical to OsHV-1 reference as well as µVar strains B and A (KY271630, KY242785.1), sequenced from France and Ireland. This finding represents the first detection of OsHV-1 DNA in a wild population of C. gigas in Argentina in association with gross mantle lesions.

A-to-I editing of Malacoherpesviridae RNAs supports the antiviral role of ADAR1 in mollusks

BackgroundAdenosine deaminase enzymes of the ADAR family are conserved in metazoans. They convert adenine into inosine in dsRNAs and thus alter both structural properties and the coding potential of their substrates. Acting on exogenous dsRNAs, ADAR1 exerts a pro- or anti-viral role in vertebrates and Drosophila.ResultsWe traced 4 ADAR homologs in 14 lophotrochozoan genomes and we classified them into ADAD, ADAR1 or ADAR2, based on phylogenetic and structural analyses of the enzymatic domain. Using RNA-seq and quantitative real time PCR we demonstrated the upregulation of one ADAR1 homolog in the bivalve Crassostrea gigas and in the gastropod Haliotis diversicolor supertexta during Ostreid herpesvirus-1 or Haliotid herpesvirus-1 infection. Accordingly, we demonstrated an extensive ADAR-mediated editing of viral RNAs. Single nucleotide variation (SNV) profiles obtained by pairing RNA- and DNA-seq data from the viral infected individuals resulted to be mostly compatible with ADAR-mediated A-to-I editing (up to 97%). SNVs occurred at low frequency in genomic hotspots, denoted by the overlapping of viral genes encoded on opposite DNA strands. The SNV sites and their upstream neighbor nucleotide indicated the targeting of selected adenosines. The analysis of viral sequences suggested that, under the pressure of the ADAR editing, the two Malacoherpesviridae genomes have evolved to reduce the number of deamination targets.ConclusionsWe report, for the first time, evidence of an extensive editing of Malacoherpesviridae RNAs attributable to host ADAR1 enzymes. The analysis of base neighbor preferences, structural features and expression profiles of molluscan ADAR1 supports the conservation of the enzyme function among metazoans and further suggested that ADAR1 exerts an antiviral role in mollusks.

Influence of environment on the pathogenesis of Ostreid herpesvirus-1 (OsHV-1) infections in Pacific oysters (Crassostrea gigas) through differential microbiome responses

The oyster microbiome is thought to contribute to the pathogenesis of mass mortality disease in Pacific oysters, associated with OsHV-1. As filter-feeders, oysters host a microbiota that can be influenced by the estuarine environment. This may alter susceptibility to OsHV-1 infections, causing variable mortality. This study aimed at: (1) differences in the microbiome of Pacific oysters with a common origin but grown in geographically distinct estuaries; (2) evaluating changes occurring in the microbiota, especially in Vibrio, and (3) differential responses of the oyster microbiome, in response to an OsHV-1 infection. Pacific oysters sourced from a single hatchery but raised separately in Patonga Creek, Shoalhaven River and Clyde River of NSW, Australia, were used and challenged with OsHV-1. The initial microbiome composition was different in the three batches and changed further, post-injection (p < 0.05). The Patonga oysters with the highest mortality also had higher OsHV-1 and Vibrio quantities compared to the other two batches (p < 0.05). The higher initial bacterial diversity in Patonga oysters decreased in moribund oysters which was not observed in the other two batches (p < 0.05). The microbiome of survivors of OsHV-1 infection and negative control oysters of two batches, did not show any changes with the relevant pre-challenged microbiome. A strong correlation was observed between the OsHV-1 and Vibrio quantities in OsHV-1 infected oysters (r = 0.6; p < 0.001). In conclusion, the Pacific oyster microbiome differed in different batches despite a common hatchery origin. Different microbiomes responded differently with a differential outcome of OsHV-1 challenge. The higher Vibrio load in oysters with higher OsHV-1 content and higher mortality, suggests a role in Vibrio in the pathogenesis of this mortality disease. This study provided insights of the potential of different estuarine environments to shape the Pacific oyster microbiome and how different microbiomes are associated with different outcomes of OsHV-1 infection.

Isolation and characterization of Vibrio harveyi as a major pathogen associated with mass mortalities of ark clam, Scapharca broughtonii, in summer

Mass mortality of ark clam, Scapharca broughtonii, occurred in a local farm along the Bohai Sea, China in August 2017. Ostreid herpesvirus SB strain, which was the only pathogen previously reported in S. broughtonii, was not detected in moribund S. broughtonii. However, four kinds of bacteria were isolated from the hepatopancreas of naturally diseased S. broughtonii, and they were further identified based on 16S rDNA gene sequence analysis. These bacteria were closely related to Vibrio harveyi (A2-1), Microbulbifer variabilis (A2-2), V. alginolyticus (A2-3), and Pseudoalteromonas sp. (A2-4). Hemolysis test showed that A2-1 exhibited a strong hemolytic activity. The cumulative mortality of S. broughtonii with foot-injected A2-1 reached 100%, which was significantly higher than that of groups infected with other bacteria. Notably, temperature showed a significant effect on the mortality of S. broughtonii infected with A2-1, as the immersion challenged test showed 100% mortality occurred at 25 ± 2 °C on the 9th day, whereas no death was observed at 15 ± 2 °C on the 12th day. Temperature also significantly affected the growth rate and extracellular protein secretion of A2-1. In conclusion, this study is the first to report that V. harveyi is a pathogen that causes mass mortality in S. broughtonii, and the temperature is an important environmental factor that affects the virulence of A2–1 on S. broughtonii.

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.

Exploring First Interactions Between Ostreid Herpesvirus 1 (OsHV-1) and Its Host, Crassostrea gigas: Effects of Specific Antiviral Antibodies and Dextran Sulfate.

Viral entry mechanisms of herpesviruses constitute a highly complex process which implicates several viral glycoproteins and different receptors on the host cell surfaces. This initial infection stage was currently undescribed for Ostreid herpes virus 1 (OsHV-1), a herpesvirus infecting bivalves including the Pacific oyster, Crassostrea gigas. To identify OsHV-1 glyproteins implicated in the attachment of the virus to oyster cells, three viral putative membrane proteins, encoded by ORF 25, 41, and 72, were selected and polyclonal antibodies against these targets were used to explore first interactions between the virus and host cells. In addition, effects of dextran sulfate, a negative charged sulfated polysaccharide, were investigated on OsHV-1 infection. Effects of antiviral antibodies and dextran sulfate were evaluated by combining viral DNA and RNA detection in spat (in vivo trials) and in oyster hemolymph (in vitro trials). Results showed that viral protein encoded by ORF 25 appeared to be involved in interaction between OsHV-1 and host cells even if other proteins are likely implicated, such as proteins encoded by ORF 72 and ORF 41. Dextran sulfate at 30 μg/mL significantly reduced the spat mortality rate in the experimental conditions. Taken together, these results contribute to better understanding the pathogenesis of the viral infection, especially during the first stage of OsHV-1 infection, and open the way toward new approaches to control OsHV-1 infection in confined facilities.

Mass mortality in Korean bay scallop (Argopecten irradians) associated with Ostreid Herpesvirus-1 μVar.

Since November 2017, mass mortalities of larvae of bay scallop (Argopecten irradians) were reported in hatcheries located at the southern area of Republic of Korea. Over 90% of larvae aged 5-10days sank to the bottom of the tank and died. The hatcheries could not produce spat, and thus artificial seed production industry incurred huge losses. We identified Ostreid Herpesvirus-1 μVar (OsHV-1 μVar) associated with mass mortality by PCR, sequencing and transmission electron microscopy (TEM). All the samples were positive for OsHV-1 μVar with 99% sequence identity to previously reported OsHV-1 μVar sequences. Partial sequence of ORF-4 of OsHV-1 detected in this study was more closely related to sequences isolated from Europe. This is the first report to confirm the mortality caused by an OsHV-1 infection in the bay scallop.

Farmer monitoring reveals the effect of tidal height on mortality risk of oysters during a herpesvirus outbreak

Abstract The intertidal zone is characterized by a sharp vertical gradient of environmental stress, which structures species distribution and their interactions. Few studies, however, have examined the influence of tidal height on host–pathogen interactions. Here, we investigated how the tidal height influence outbreak of the Ostreid herpesvirus type 1 (OsHV-1) affecting the Pacific oyster. A volunteer network composed of 20 oyster growers monitored the survival of 28 batches of oysters during an epizootic event in Southern Brittany, France. Oysters were spat from wild collection or hatchery production. The sampling sites were spread over a 150-km2 area with a tidal height ranging from 0.98 to 2.90 m. Concomitantly, we followed survival of oyster spats in relation with OsHV-1 DNA detection at two sites and conducted risk analysis. We found that tidal height was associated with a lower risk of mortality. This effect was higher for hatchery than for wild oysters probably reflecting differences in health status. Our study opens perspectives for mitigation strategies based on tidal height and emphasizes the value of volunteer science in marine epidemiological studies.

Prior exposure to Ostreid herpesvirus 1 (OsHV-1) at 18 °C is associated with improved survival of juvenile Pacific oysters (Crassostrea gigas) following challenge at 22 °C

The high mortality and economic loss in farmed Pacific oysters (Crassostrea gigas) caused by Ostreid herpesvirus 1 (OsHV-1) has spurred research into strategies to mitigate the impact of the disease. Water temperature strongly influences the outcome of exposure with 14 °C not being permissive for an epidemic, infection occurring with limited mortality at 18 °C if there is a high viral dose, and full disease expression occurring at 22 °C. Observations in the field suggest improved survival to subsequent exposures in cohorts of oysters that have survived an OsHV-1 outbreak, but this assessment is complicated by the increasing age of the oysters, different viral exposures, different environmental conditions and the removal of naturally susceptible individuals. A laboratory infection model was used to evaluate the effect on survival of two temporally-spaced challenges with OsHV-1 under different water temperature regimes. Triploid Pacific oyster spat were recruited from commercial farm stocks at 6 months of age and exposed to OsHV-1 by intramuscular injection at 18 °C or 22 °C and then re-exposed at either 18 °C or 22 °C. Mortality did not occur in oysters exposed at 18 °C or when the temperature was increased to 22 °C at 14 days post challenge. Oysters challenged with OsHV-1 at 22 °C were protected if pre-exposed to OsHV-1 at 18 °C (Hazard ratio: 0.22, 95% CI: 0.06–0.80). The present study suggests that non-lethal exposure to OsHV-1 can reduce the mortality on subsequent exposure to the virus, but further experiments are required to investigate the duration of the protective response and its effect in different environments. Such protection may help describe the dynamics of seasonally recurrent disease epidemics and provide a novel approach to disease management.

Variability in the Composition of Pacific Oyster Microbiomes Across Oyster Families Exhibiting Different Levels of Susceptibility to OsHV-1 μvar Disease.

Oyster diseases are a major impediment to the profitability and growth of the oyster aquaculture industry. In recent years, geographically widespread outbreaks of disease caused by ostreid herpesvirus-1 microvariant (OsHV-1 μvar) have led to mass mortalities among Crassostrea gigas, the Pacific Oyster. Attempts to minimize the impact of this disease have been largely focused on breeding programs, and although these have shown some success in producing oyster families with reduced mortality, the mechanism(s) behind this protection is poorly understood. One possible factor is modification of the C. gigas microbiome. To explore how breeding for resistance to OsHV-1 μvar affects the oyster microbiome, we used 16S rRNA amplicon sequencing to characterize the bacterial communities associated with 35 C. gigas families, incorporating oysters with different levels of susceptibility to OsHV-1 μvar disease. The microbiomes of disease-susceptible families were significantly different to the microbiomes of disease-resistant families. OTUs assigned to the Photobacterium, Vibrio, Aliivibrio, Streptococcus, and Roseovarius genera were associated with low disease resistance. In partial support of this finding, qPCR identified a statistically significant increase of Vibrio-specific 16S rRNA gene copies in the low disease resistance families, possibly indicative of a reduced host immune response to these pathogens. In addition to these results, examination of the core microbiome revealed that each family possessed a small core community, with OTUs assigned to the Winogradskyella genus and the Bradyrhizobiaceae family consistent members across most disease-resistant families. This study examines patterns in the microbiome of oyster families exhibiting differing levels of OsHV-1 μvar disease resistance and reveals some key bacterial taxa that may provide a protective or detrimental role in OsHV-1 μvar disease outbreaks.

Synthesis of the “PLAN DE SAUVEGARDE” using selected all-triploid oysters to reduce the shortage of spat in France due to OsHV-1–associated mortality in Crassostrea gigas

Due to massive mortality of Crassostrea gigas spat in France since 2008, a “plan de sauvegarde” was set up from 2011 to 2014 (hereafter referred to as PS1 to PS4), in order to reduce the shortage of spat. This plan involved the participation of commercial hatcheries, the French Research Institute for Exploitation of the Sea (Ifremer), and the Direction des Pêches Maritimes et de l'Aquaculture (DPMA) of the French Ministry of Agriculture. It was based on selecting diploid lines of C. gigas for their higher resistance to the oyster herpesvirus OsHV-1 (2nR group), and one of these lines was subsequently tetraploidized (4nR group). Both the 2nR and 4nR groups were produced by Ifremer, and then transferred to commercial hatcheries that produced the selected triploids (3nR groups). We report here the mortality rates of the 3nR group for each of the four “plan de sauvegarde” campaigns and compare these with the mortalities of the classic production of commercial hatcheries (both 2n and 3n), benchmarks of selected (2nR group) and unselected (2n-control group) oysters produced by Ifremer, and wild-caught spat, representing a total of 104 diploid and triploids batches. For PS1, the 3nR group had a mean mortality of 67% and did not show any advantage over the 2n- and 3n-commercial groups, suggesting a lack of genetic progress in the 2nR and 4nR groups. For PS2, OsHV-1 resistance was increased in both the 2nR and 4nR groups and, consequently, the 3nR group exhibited a mean mortality of 52%, which was significantly lower than the mortality of the 2n- (87%) and 3n-(76%) commercial groups in 2012. Unfortunately, the mortality of the 3nR group reached 62% and 71% in PS3 and PS4, respectively, although it was expected to be lower than that in PS2. OsHV-1 DNA was quantified in the live oysters at deployment (1356 oysters) and at the endpoint (1171 oysters), as well as in moribund oysters sampled during peak mortality (539 oysters). The results strongly supported the involvement of this pathogen during the main mortality outbreak in May/June. Meanwhile, Vibrio aestuarianus was also suspected to cause unexpected mortality of PS3 oysters in August and September, and it was detected in moribund PS4 oysters during both the mortality events, in May and July. Despite genetic improvement for OsHV-1 resistance, this translated into variable commercial genetic gain. This could be explained by the limited genetic backgrounds of the 2nR and 4nR groups, the reemergence of V. aestuarianus in France since 2012, the changing levels of genetic improvement in both the 3nR group and the commercial groups, as well as the limited broodstock genetic variation where small numbers of males were used. Results on growth and yield are discussed.