Oyster Mortality Research Articles

A literature review as an aid to identify strategies for mitigating ostreid herpesvirus 1 in Crassostrea gigas hatchery and nursery systems

AbstractAn understanding of husbandry strategies and any associated risk factors is important for designing management control measures that can reduce mortality in Pacific oysters, Crassostrea gigas, caused by ostreid herpesvirus 1 (OsHV‐1). The type of culture facility can be considered in relation to the potential pathways that could lead to the entry of a pathogen and its survival. In addition, the animal host (e.g. age, physiological state, selective breeding programmes), husbandry procedures (e.g. stocking density), the pathogen itself (e.g. pathogenicity, virulence) and environmental effects (e.g. temperature) represent other relevant interconnected factors. However, all these factors provide valuable background information for outlining the mitigation strategies needed by the industry, as well as in the context of surveillance and biosecurity programmes. These control mechanisms for hatchery or nursery areas are related to movement restrictions, water treatment, virus inactivation, the production calendar and practical farm management decisions. This comprehensive literature review compiles information related to such approaches and also includes the different existing guidelines suggested for control of OsHV‐1. Therefore, the review represents a solid foundation for a more critical appraisal currently being developed to support recommendations for disease management strategies.

Inefficient immune response is associated with microbial permissiveness in juvenile oysters affected by mass mortalities on field

Since 2008, juvenile Crassostrea gigas oysters have suffered from massive mortalities in European farming areas. This disease of complex etiology is still incompletely understood. Triggered by an elevated seawater temperature, it has been associated to infections by a herpes virus named OsHV-1 as well as pathogenic vibrios of the Splendidus clade. Ruling out the complexity of the disease, most of our current knowledge has been acquired in controlled experiments. Among the many unsolved questions, it is still ignored what role immunity plays in the capacity oysters have to survive an infectious episode. Here we show that juvenile oysters susceptible to the disease mount an inefficient immune response associated with microbial permissiveness and death. We found that, in contrast to resistant adult oysters having survived an earlier episode of mortality, susceptible juvenile oysters never exposed to infectious episodes died by more than 90% in a field experiment. Susceptible oysters were heavily colonized by OsHV-1 herpes virus as well as bacteria including vibrios potentially pathogenic for oysters, which proliferated in oyster flesh and body fluids during the mortality event. Nonetheless, susceptible oysters were found to sense microbes as indicated by an overexpression of immune receptors and immune signaling pathways. However, they did not express important immune effectors involved in antimicrobial immunity and apoptosis and showed repressed expression of genes involved in ROS and metal homeostasis. This contrasted with resistant oysters, which expressed those important effectors, controlled bacterial and viral colonization and showed 100% survival to the mortality event. Altogether, our results demonstrate that the immune response mounted by susceptible oysters lacks some important immune functions and fails in controlling microbial proliferation. This study opens the way to more holistic studies on the “mass mortality syndrome”, which are now required to decipher the sequence of events leading to oyster mortalities and determine the relative weight of pathogens, oyster genetics and oyster-associated microbiota in the disease.

Metabolism of the Pacific oyster, Crassostrea gigas, is influenced by salinity and modulates survival to the Ostreid herpesvirus OsHV-1.

ABSTRACTThe Pacific oyster, Crassostrea gigas, is an osmoconforming bivalve exposed to wide salinity fluctuations. The physiological mechanisms used by oysters to cope with salinity stress are energy demanding and may impair other processes, such as defense against pathogens. This oyster species has been experiencing recurrent mortality events caused by the Ostreid herpesvirus 1 (OsHV-1). The objectives of this study were to investigate the effect of salinity (10, 15, 25 and 35‰) on energetic reserves, key enzyme activities and membrane fatty acids, and to identify the metabolic risk factors related to OsHV-1-induced mortality of oysters. Acclimation to low salinity led to increased water content, protein level, and energetic reserves (carbohydrates and triglycerides) of oysters. The latter was consistent with lower activity of hexokinase, the first enzyme involved in glycolysis, up-regulation of AMP-activated protein kinase, a major regulator of cellular energy metabolism, and lower activity of catalase, an antioxidant enzyme involved in management of reactive oxygen species. Acclimation to salinity also involved a major remodeling of membrane fatty acids. Particularly, 20:4n-6 decreased linearly with decreasing salinity, likely reflecting its mobilization for prostaglandin synthesis in oysters. The survival of oysters exposed to OsHV-1 varied from 43% to 96% according to salinity (Fuhrmann et al., 2016). Risk analyses showed that activity of superoxide dismutase and levels of proteins, carbohydrates, and triglycerides were associated with a reduced risk of death. Therefore, animals with a higher antioxidant activity and a better physiological condition seemed less susceptible to OsHV-1.

Size-dependent predation and intraspecific inhibition of an estuarine snail feeding on oysters

Predator outbreaks have increased in the past two decades in many ecosystems and are predicted to become more common with climate change. During these outbreaks, predator densities increase rapidly, and can cause large reductions in prey populations or shifts in prey size structure. However, unexpected interactions may occur at high predator densities, necessitating a mechanistic understanding of how increased predator density affects predator-prey dynamics. In the northern Gulf of Mexico, outbreaks of southern oyster drill, Stramonita haemastoma (Linnaeus, 1767), occur during high salinity events, and can greatly reduce eastern oyster, Crassostrea virginica (Gmelin, 1791), populations. A large outbreak of drills occurred from 2013 to 2015 in Apalachicola Bay, FL which corresponded with an oyster fishery collapse in that bay. To improve our mechanistic understanding of predation during such a high-density outbreak, laboratory experiments based on field observations were used to quantify the prey size selection by drills and the drill functional response, as a function of drill abundance. Drills fed on medium-sized oysters (50–75 mm) more often than small and larger-sized oysters, and often formed aggregations during feeding events. However, despite this aggregative response, there was a negative relationship between per capita feeding rates and drill abundance. Indeed, the Crowley-Martin functional response model had the most parsimonious fit to the data, suggesting that predator-predator inhibition reduced attack rates and increased handling times. Due to an increase in regional drought conditions and water usage in the southeastern United States, drill outbreaks will likely increase in frequency and duration. A greater understanding of how predation rates change with predator densities during outbreaks will improve predictions of oyster mortality, and strengthen the scientific framework for oyster fishery decisions.

Refining the tethering of American oysters (Crassostrea virginica) to measure the effects of two environmental stressors

Tethering assays, or the physical restraint of test organisms, has been used in the past to measure selected organisms' response to stressors while removing the observer from the experimental setting. Although informative for monitoring and hypothesis testing, these assays often used microfilaments that have been found to be too invasive or prone to biases given their effects on test organisms' behavior. Here, we describe a new variation of tethering using American oysters (Crassostrea virginica) and illustrate its use in the study of their mortality rates as a result of two stressors: siltation and predation by a non-indigenous species. Our protocol identified a resistant (non-toxic) glue that could be used to attach oysters to stone slabs, thus partially mimicking the natural cementation of the shell to natural substrates. This variation of tethering was harmless and maintained oysters' body position and natural ability to filter feed. Using tethered oysters in separate two-week field cage experiments, we also show how siltation and predation by a non-indigenous species (the European green crab, Carcinus maenas), caused a gradual, easily measurable increase in oyster mortality rates. We argue that this variation of tethering is a cost-effective and advantageous way to monitor or test the effects of these and other stressors on oysters and similar species.

Pelagic oxycline and damage potential of hypoxia to the Pacific oyster Crassostrea gigas suspended in longline aquaculture systems

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 9:461-468 (2017) - DOI: https://doi.org/10.3354/aei00245 Pelagic oxycline and damage potential of hypoxia to the Pacific oyster Crassostrea gigas suspended in longline aquaculture systems Sang Jun Lee1, Qtae Jo1,*, Jong-Cheol Han2, Yeong-Cheol Park3, Tae Gyu Park4 1Shellfish Aquaculture Lab, South Sea Fisheries Research Institute, NIFS, Jeonnam 59780, South Korea 2Shellfish Hatchery Center, Southeast Sea Fisheries Research Institute, NIFS, Gyeognam 52440, South Korea 3Environmental Analysis Lab, Southeast Sea Fisheries Research Institute, NIFS, Gyeognam 53085, South Korea 4Harmful Algal Bloom Lab, Southeast Sea Fisheries Research Institute, NIFS, Gyeognam 53085, South Korea *Corresponding author: qtjo@korea.kr ABSTRACT: Depletion in dissolved oxygen (DO) occurred in 2 semi-enclosed shallow bays in Korea in which Crassostrea gigas were suspended for aquaculture. DO depletion started at the bottom and expanded rapidly upwards for days until it was blocked by a temperature-driven density differential. This resulted in the formation of pelagic oxyclines at a depth of 2–4 m in one bay (Hypoxic Site 1, HS-1) and 2–5 m in the other (Hypoxic Site 2, HS-2). Water above the oxycline was normoxic (>4 mg l-1), while water below the oxycline was hypoxic (<1 mg l-1). The oxyclines were accompanied by significant changes in environmental variables and phytoplankton composition, but these changes were not fatal to the oysters. However, the oxyclines themselves caused oyster mortality: complete mortality below the oxyclines and depth-dependent mortality within the oxycline. Interestingly, mortality was observed in a significant number of oysters above the oxyclines compared with the reference site, and greater mortality was observed in HS-1 than HS‑2. These findings suggest the existence of toxic compounds that diffuse up from below the oxycline in shallow waters and exert effects that overshadow those of DO. The higher mortalities in the HS-1 normoxic layer supported this influence from the bottom layer. In a subsequent experiment, we observed additional mortalities among the surviving oysters in the upper normoxic waters after the bays had completely reoxygenated during fall turnover. These data provide useful observations of hypoxia in highly sheltered shallow waters and can be used to guide site selection for oyster longline aquaculture. KEY WORDS: Oxycline · Sheltered shallow coast · Oyster farm · Damage Full text in pdf format PreviousCite this article as: Lee SJ, Jo Q, Han JC, Park YC, Park TG (2017) Pelagic oxycline and damage potential of hypoxia to the Pacific oyster Crassostrea gigas suspended in longline aquaculture systems. Aquacult Environ Interact 9:461-468. https://doi.org/10.3354/aei00245 Export citation RSS - Facebook - Tweet - linkedIn Cited by Published in AEI Vol. 9. Online publication date: November 22, 2017 Print ISSN: 1869-215X; Online ISSN: 1869-7534 Copyright © 2017 Inter-Research.

Local and regional stressors interact to drive a salinization‐induced outbreak of predators on oyster reefs

AbstractPredator outbreaks are predicted to increasingly decimate economically and ecologically important prey populations because global climate change and food‐web modifications frequently facilitate predators and stress prey. Natural systems are organized hierarchically, with processes operating at multiple scales giving rise to patterns of biodiversity, so predicting and managing outbreaks requires a framework that accounts for the effects of both local and regional stressors. Here, we used the comparative experimental approach to investigate whether the collapse of a nationally important oyster fishery in the Gulf of Mexico (Apalachicola Bay, Florida) could have been (1) caused proximally by a predator outbreak and (2) whether this outbreak was mediated by local‐ and/or regional‐scale forces. During the fishery collapse, we paired experiments with monitoring in Apalachicola Bay and found elevated water salinity, high abundance of predatory snails, and intense oyster mortality due to predation. By repeating these experiments over 4 yr, we found that periods of reduced water salinity inhibited predation on oysters. To partition the influence of local‐versus‐regional factors on this predator outbreak, we simultaneously replicated the paired experiments and monitoring in a nearby bay (Ochlockonee Bay) that shares the same regional‐scale rainfall conditions. Increasing freshwater withdrawals from the watershed that drains into Apalachicola Bay have increased salinities in that bay, but there have not been similar withdrawals in the Ochlockonee Bay watershed. Therefore, Apalachicola Bay experienced a localized anthropogenic stress, while both bays experienced regional stress from drought. In Ochlockonee Bay, our experiments demonstrated that the river maintained sufficiently low salinity to provide ~50% of oyster reefs with a refuge from predation. In contrast, salinity‐dependent predation in Apalachicola Bay extended up to the river mouth. Given the stark differences in upstream water withdrawals between these watersheds, it is reasonable to surmise that these withdrawals exacerbated the stress of regional drought, created the difference in predation between the two bays, and thus may have precipitated the oyster fishery collapse. Our study provides empirical support for recent theory about the hierarchical organization of ecosystems, which predicts that stressors will interact across scales to cause localized predator outbreaks.

First description of a mortality event in adult Pacific oysters in Italy associated with infection by a Tenacibaculum soleae strain.

Summer mortality episodes in adult Pacific oysters have been described since the 1950s in various farming areas. Starting in 2012, a recrudescence of mortalities in commercial-sized oysters was first observed in France and then in Italy, with seasonality extension and translation later in the year. Moribund individuals collected during an event in Italy in December 2014 showed yellowish lesions of the mantle and adductor muscle. Histological examination revealed filamentous bacteria associated with necrotic areas. Quantitative PCRs targeting OsHV-1 and Vibrio aestuarianus detected only high loads of the pathogenic bacteria in tissues of symptomatic individuals. A lower diversity of the hemolymph microbiota was also evidenced in moribund individuals, with a predominance of Vibrio and Arcobacter species. A strain of Flavobacteriaceae was isolated from all the symptomatic individuals. Sequence analysis of the 16S rRNA gene identified the strain as Tenacibaculum soleae. When strain pathogenicity was tested by injection in adult individuals, it induced mortality rates of up to 45%, even in the absence of V.aestuarianus. As mortality occurred only 11days post-infection, further investigation is needed to determine its effective virulence in natural conditions. This is the first description of a Tenacibaculum strain associated with bivalve mortalities.

Timing of stressors alters interactive effects on a coastal foundation species.

The effects of climate-driven stressors on organismal performance and ecosystem functioning have been investigated across many systems; however, manipulative experiments generally apply stressors as constant and simultaneous treatments, rather than accurately reflecting temporal patterns in the natural environment. Here, we assessed the effects of temporal patterns of high aerial temperature and low salinity on survival of Olympia oysters (Ostrea lurida), a foundation species of conservation and restoration concern. As single stressors, low salinity (5 and 10psu) and the highest air temperature (40°C) resulted in oyster mortality of 55.8, 11.3, and 23.5%, respectively. When applied on the same day, low salinity and high air temperature had synergistic negative effects that increased oyster mortality. This was true even for stressor levels that were relatively mild when applied alone (10psu and 35°C). However, recovery times of two or four weeks between stressors eliminated the synergistic effects. Given that most natural systems threatened by climate change are subject to multiple stressors that vary in the timing of their occurrence, our results suggest that it is important to examine temporal variation of stressors in order to more accurately understand the possible biological responses to global change.

Contrasting complexity of adjacent habitats influences the strength of cascading predatory effects

Although cascading effects of top predators can help structure communities, their influence may vary across habitats that differentially protect prey. Therefore, to understand how and to what degree habitat complexity can affect trophic interactions in adjacent habitats, we used a combination of a broad regional-scale survey, manipulative field trials, and an outdoor mesocosm experiment to quantify predator-prey interaction strengths across four trophic levels. Within estuaries of the southeastern USA, bonnethead sharks (Sphyrna tiburo) hunt blue crabs on mudflats and adjacent oyster reefs, two habitats with vastly different aboveground structure. Using 12-h tethering trials of blue crabs we quantified habitat-dependent loss rates of 37% on reefs and 78% on mudflats. We hypothesized that the sharks' predatory effects on blue crabs would cascade down to release a lower-level mud crab predator, which subsequently would increase juvenile oyster mortality, but that the cascade strength would be habitat-dependent. We experimentally manipulated predator combinations in split-plot mesocosms containing reef and mudflat habitats, and quantified oyster mortality. Bonnetheads exerted strong consumptive and non-consumptive effects on blue crabs, which ceased eating oysters in the sharks' presence. However, mud crabs, regardless of shark and blue crab presence, continued to consume oysters, especially within the structural refuge of the reef where they kept oyster mortality high. Thus, bonnetheads indirectly boosted oyster survival, but only on the mudflat where mud crabs were less active. Our work demonstrates how structural differences in adjacent habitats can moderate trophic cascades, particularly when mesopredators exhibit differential use of structure and different sensitivities to top predators.

Oyster (Crassostrea virginica [Gmelin, 1791]) Mortality at Prolonged Exposures to High Temperature and Low Salinity

ABSTRACT Mortality of two size classes ( 35 mm) of eastern oysters Crassostrea virginica when exposed to combinations of low salinity (1, 2, 3, and 4) for extended periods (up to 30 days) at summer water temperatures typical of the Virginia Chesapeake Bay subestuaries was examined. A critical salinity-temperature combination of less than two at greater than 28°C for more than 1 wk exposure for oyster mortality is suggested. A review of limited historical salinity-temperature tolerance data suggest selection of local populations of oysters having differing salinity tolerances. Such selection may prove critical to persistence of low-salinity populations in the Chesapeake Bay subestuaries with projected climate change.

Effects of Seeding Date and Seed Size on Crassostrea gigas (Thunberg, 1793) Culture in a Subtropical Climate

Studies that carefully monitor and attempt to explain causes of massive oyster mortalities are urgently needed. The objective of the present study was to assess the effects of seeding dates on the mortality and reproductive cycle of the oyster Crassostrea gigas. Seeds of two sizes (3.5–4 mm and 1.5–2 mm) were planted during three seasons: early summer, early autumn, and late autumn. The experimental period was 6 mo. Mortalities of C. gigas at culture phase nursery lantern-nets, intermediate lantern-nets, and final lantern-nets were recorded. The reproductive cycle of the oysters grown from 3.5 to 4 mm seeds planted in each season was closely monitored (bimonthly) during the experimental period. In addition, seawater parameters [temperature (daily) and salinity (weekly)] were monitored throughout the experimental period. Results of this study demonstrated that the planting season affected C. gigas mortality after the nursery phase. Lower cumulative mortality was observed for 3.5–4 mm seeds than for 1.5–2 mm seeds when the seeds were planted in early (MarL: 0.29 ± 0.02) and late (JunL: 0.22 ± 0.10) autumn, after 6 mo of culture. The reproductive cycle of seeds planted in the summer exhibited an early maturation process, and those planted in the autumn exhibited late reproductive development.

Effects of controlled air exposure on the survival, growth, condition, pathogen loads and refrigerated shelf life of eastern oysters

The benefits of exposing eastern oysters to air during commercial culture have not been well-characterized. An adjustable longline system (ALS) designed in Australia and recently adopted by the nascent aquaculture industry in the northern Gulf of Mexico, allows growing oysters at any position in the water column and is perfectly suited to study the benefits of air exposure. Four-month old diploid oysters were deployed in an ALS and divided into three groups: 1) oysters exposed to air daily for 8–12 hr during low tide, 2) oysters exposed to air weekly (~24 hr once a week), and 3) oysters kept subtidally. Oyster mortality and growth rates, Perkinsus marinus load and condition index were then determined every 3 months over 2 years, while refrigerated shelf life and Vibrio vulnificus load were determined in summer and early fall of the second year. Summer mortalities were delayed, P. marinus infection intensities tended to be lower and condition index was significantly higher in oysters exposed to air daily compared with oysters exposed to air weekly or held subtidally. Shell heights of oysters exposed to air daily were lower for most of the study due to a lower growth rate during the initial sampling interval following deployment. No consistent differences were found in V. vulnificus loads or refrigerated shelf lives between the groups. It is recommended that ALS be set so that oysters are kept subtidally fall through spring to promote growth, and exposed to air daily during summer to delay P. marinus proliferation.

Exploring the decline of oyster beds in Atlantic Canada shorelines: potential effects of crab predation on American oysters (Crassostrea virginica)

Atlantic Canada’s American oyster (Crassostrea virginica) beds, while economically and ecologically important, have been in decline over the past few decades. Predation by crabs, in particular by the European green crab (Carcinus maenas), has been proposed as one of the potential causes of such decline. Hence, this study examined oyster mortality levels in multiple beds across Prince Edward Island (PEI) and then experimentally assessed the contribution of green crab predation to oyster mortality. Results from surveys conducted in 10 estuaries across PEI in 2014 indicate that the probability of mortality for small oysters was significantly higher when green crabs were present then in areas without green crabs. This probability of mortality was significantly less when there was the presence of alternative prey like natural mussel beds (Mytilus edulis). The odds of oyster mortality were also higher when beds had rock crabs (Cancer irroratus) compared to beds with no rock crabs. Given the potential importance of green crab predation, its influence was assessed in 2015 using two field experiments with tethered oysters. Our results indicate that odds of small oyster mortality occurring were much higher in green crab inclusion cages than in the open environment and the exclusion cages. These results reaffirm that oysters up to ~40 mm SL are vulnerable to predation, and at least some of the mortality affecting these oysters can be causally attributed to green crab predation. Green crab predation rates upon small oysters are relevant given the economic benefits and ecosystem services provided by these bivalves. They highlight the need for the industry to consider mitigation measures and potentially adapt their oyster growing strategies.

Impact of dispersant on early life stages of the water flea Daphnia magna and the eastern oyster Crassostrea virginica.

In response to the 2010 Deepwater Horizon oil spill, over 1 million gallons of dispersant were applied in Gulf of Mexico offshore waters; Corexit 9500 was the most applied dispersant. The impact on organisms in nearshore and freshwaters has received little scrutiny. Acute 48 h toxicity of Corexit 9500 and a new hyperbranched polyethylenimine (HPEI) dispersant-like compound were evaluated for the freshwater indicator organism, Daphnia magna and for larval and early spat stages of the Eastern oyster, Crassostrea virginica. For D. magna, Corexit 9500 demonstrated toxicity (EC50 of 0.14 [0.13, 0.15] ppm) similar to the 10-kDa HPEI (EC50 of 0.16 [0.12, 0.19] ppm). HPEI toxicity increased as a function of molecular weight (1.2 to 750kDa). The 10 kDa size HPEI was further investigated because it dispersed crude oil with equal effectiveness as Corexit. For Corexit, 100% oyster mortality was detected for the ≤0.2-mm size classes and mortality >50% for the 0.3- and 0.7-mm size classes at the two greatest concentrations (25 and 50ppm). HPEI (10kDa) exhibited low mortality rates (<30%) for all concentrations for all oyster size classes except the 0.1-mm class. Although mortality rates for this size class were up to 60%, mortality was still less than the mortality caused by Corexit 9500. The low toxicity of HPEI polymers for C. virginica in comparison with Corexit 9500 suggests that HPEI polymers warrant further study.

Threat of Predation Does Not Affect Crassostrea virginica Filtration

Biotic interactions can structure ecological communities and influence ecosystem functioning. As ecosystem engineers and filter feeders, bivalves often have disproportionately large effects on ecosystem functioning. They also utilize numerous morphological and behavioral responses to reduce predation, which can include changes in their filtration rates. To test the response of Crassostrea virginica filtration rates to the presence of predators, juvenile and adult oysters were separately exposed to varying types of predation risk from Callinectes sapidus and Panopeus herbstii in outdoor mesocosms. Water column chlorophyll a concentrations and crab behavior were measured over the duration of the experiment. Predation risk had no effect on oyster reef drawdown of chlorophyll a, which suggests that this important ecosystem function of oyster reefs is not mediated by behaviorally induced predator effects. Therefore, efforts to model how oyster predators influence filtration rates and associated ecosystem services should focus primarily on the factors that influence oyster mortality rather than predator effects on oyster behavior.

Increasing microhabitat complexity on seawalls can reduce fish predation on native oysters

Increasingly, urbanised coastlines are being armoured by shoreline protection structures, such as seawalls. Seawalls typically lack the complex microhabitats and protective spaces of natural shorelines and consequently organisms that settle on them may be particularly susceptible to predation. We tested whether the addition of complex microhabitats to seawalls enhances the survivorship of oysters, key habitat-forming species on intertidal shores, by reducing the intensity of predation. At two sites in Sydney Harbour, we compared the magnitude and sources of mortality of juvenile oysters among (1) flat tiles, without crevices or ridges; (2) complex tiles, with 2.5cm high ridges, separated by crevices; and (3) complex tiles, with 5cm high ridges, separated by crevices. We also compared predatory fish visitation and feeding among sites and treatments using GoPro® cameras. The abundance and feeding of predatory fish was much greater at one of the study sites than the other, but at neither site differed among treatments. At the site with greater predatory fish abundances, survivorship of juvenile oysters was 50% greater on the 5cm complex tiles than flat tiles, and on complex tiles approximately 300% greater in crevices than on ridges. Of the dead oysters, almost all were cracked, indicative of fish predation. In contrast, at the site with fewer predatory fish, there were no detectable differences in oyster survivorship between treatments. These results suggest that the addition of complex habitat to seawalls could be an effective strategy in reducing fish predation pressure on juvenile oysters at sites with abundant predatory fish. A greater understanding of the site-specific pressures is required to enhance the abundances of desirable species and functions on seawalls.

Haemocytes collected from experimentally infected Pacific oysters, Crassostrea gigas: Detection of ostreid herpesvirus 1 DNA, RNA, and proteins in relation with inhibition of apoptosis.

Recent transcriptomic approaches focused on anti-viral immunity in molluscs lead to the assumption that the innate immune system, such as apoptosis, plays a crucial role against ostreid herpesvirus type 1 (OsHV-1), infecting Pacific cupped oyster, Crassostrea gigas. Apoptosis constitutes a major mechanism of anti-viral response by limiting viral spread and eliminating infected cells. In this way, an OsHV-1 challenge was performed and oysters were monitored at three times post injection to investigate viral infection and host response: 2h (early after viral injection in the adductor muscle), 24h (intermediate time), and 48h (just before first oyster mortality record). Virus infection, associated with high cumulative mortality rates (74% and 100%), was demonstrated in haemocytes by combining several detection techniques such as real-time PCR, real-time RT PCR, immunofluorescence assay, and transmission electron microscopy examination. High viral DNA amounts ranged from 5.46×104 to 3.68×105 DNA copies ng-1 of total DNA, were detected in dead oysters and an increase of viral transcripts was observed from 2, 24, and 48hpi for the five targeted OsHV-1 genes encoding three putative membrane proteins (ORFs 25, 41, and 72), a putative dUTPase (ORF 75), and a putative apoptosis inhibitor (ORF 87). Apoptosis was studied at molecular and cellular levels with an early marker (phosphatidyl-serine externalisation measured by flow cytometry and epifluorescence microscopy) and a later parameter (DNA fragmentation by terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling assay (TUNEL)). The down-regulation of genes encoding proteins involved in the activation of the apoptotic pathway (TNF and caspase 3) and the up-regulation of genes encoding anti-apoptotic proteins (IAP-2, and Bcl-2) suggested an important anti-apoptosis phenomenon in haemocytes from OsHV-1 infected oysters at 24 and 48hpi. Additionally, more phosphatidyl-serines were externalized and more cells with DNA fragmentation were observed in haemocytes collected from artificial seawater injected oysters than in haemocytes collected from OsHV-1 infected oysters at 24 and 48hpi, suggesting an inhibition of the apoptotic process in presence of the virus. In conclusion, this study is the first to focus on C. gigas haemocytes, cells involved in the host immune defense, during an OsHV-1 challenge in controlled conditions by combining various and original approaches to investigate apoptosis at molecular and cellular levels.

Influence of OSHV-1 oyster mortality episode on dissolved inorganic fluxes: An ex situ experiment at the individual scale

Ostreid herpesvirus 1 (OsHV-1 μvar) infection has caused significant mortalities in juvenile oysters (Crassostrea gigas). In contrast to the practices of other animal production industries, sick and dead oysters are not separated from live ones and are left to decay in the surrounding environment, with unknown consequences on fluxes of dissolved materials. A laboratory approach was used in this study to test the influence of oyster mortality episode on dissolved inorganic fluxes at the oyster interface, dissociating (i) the effect of viral infection on metabolism of juvenile oysters and (ii) the effect of flesh decomposition on oxygen consumption and nutrient releases at the individual scale. Nine batches of juvenile oysters (Individual Total wet weight 1g) were infected via injection of OsHV-1 enriched inoculums at different viral loads (108 and 109 OsHV-1 DNA copies per oyster) to explore infection thresholds. Oysters injected with filtered seawater were used as controls (C). Oysters were maintained under standard conditions to avoid stress linked to hypoxia, starvation, or ammonia excess. Before, after the injection and during the mortality episode, i.e. at days 1, 3, 7, 10 and 14, nine oysters per treatment were incubated in individual metabolic chambers to quantify oxygen, ammonium and phosphate fluxes at the seawater-oyster interface. Nine empty chambers served as a reference. Injections of the two viral loads of OsHV-1 induced similar mortality rates (38%), beginning at day 3 and lasting until day 14. The observed mortality kinetics were slower than those reported in previous experimental pathology studies, but comparable to those observed in the field (Thau lagoon, France). This study highlights that oxygen and nutrient fluxes significantly varied during mortality episode. Indeed (i) OsHV-1 infection firstly modifies oyster metabolism, with significant decreases in oxygen consumption and ammonium excretion, and (ii) dead oysters lead to a strong increase of ammonium (6 fold) and phosphate (41 fold) fluxes and a decrease in the N/P ratio due to mineralisation of their flesh. The latter may modify the structure of the planktonic community in the field during mortality episode. This study is a first step of the MORTAFLUX program. The second step was to in situ confirm this abnormal nutrient loading during a mortality episode and show its impact on bacterio-, phyto- and protozoo-plankton.

Non-indigenous predators threaten ecosystem engineers: Interactive effects of green crab and oyster size on American oyster mortality

Non-indigenous green crabs (Carcinus maenas) are emerging as important predators of autogenic engineers like American oysters (Crassostrea virginica) throughout the eastern seaboard of Canada and the United States. To document the spreading distribution of green crabs, we carried out surveys in seven sites of Prince Edward Island during three fall seasons. To assess the potential impact of green crabs on oyster mortality in relation to predator and prey size, we conducted multiple predator-prey manipulations in the field and laboratory. The surveys confirmed an ongoing green crab spread into new productive oyster habitats while rapidly increasing in numbers in areas where crabs had established already. The experiments measured mortality rates on four sizes of oysters exposed to three sizes of crab, and lasted 3–5 days. The outcomes of experiments conducted in Vexar® bags, laboratory tanks and field cages were consistent and were heavily dependent on both crab size and oyster size: while little predation occurred on large oysters, large and medium green crabs preyed heavily on small sizes. Oysters reached a refuge within the 35–55 mm shell length range; below that range, oysters suffered high mortality due to green crab predation and thus require management measures to enhance their survival. These results are most directly applicable to aquaculture operations and restoration initiatives but have implications for oyster sustainability.