Plastics are persistent synthetic polymers that accumulate as waste in the marine environment. Microplastic (MP) particles are derived from the breakdown of larger debris or can enter the environment as microscopic fragments. Because filter-feeder organisms ingest MP while feeding, they are likely to be impacted by MP pollution. To assess the impact of polystyrene microspheres (micro-PS) on the physiology of the Pacific oyster, adult oysters were experimentally exposed to virgin micro-PS (2 and 6 μm in diameter; 0.023 mg·L −1 ) for 2 mo during a reproductive cycle. Effects were investigated on ecophysiological parameters; cellular, transcriptomic, and proteomic responses; fecundity; and offspring development. Oysters preferentially ingested the 6-μm micro-PS over the 2-μm-diameter particles. Consumption of microalgae and absorption efficiency were significantly higher in exposed oysters, suggesting compensatory and physical effects on both digestive parameters. After 2 mo, exposed oysters had significant decreases in oocyte number (−38%), diameter (−5%), and sperm velocity (−23%). The D-larval yield and larval development of offspring derived from exposed parents decreased by 41% and 18%, respectively, compared with control offspring. Dynamic energy budget modeling, supported by transcriptomic profiles, suggested a significant shift of energy allocation from reproduction to structural growth, and elevated maintenance costs in exposed oysters, which is thought to be caused by interference with energy uptake. Molecular signatures of endocrine disruption were also revealed, but no endocrine disruptors were found in the biological samples. This study provides evidence that micro-PS cause feeding modifications and reproductive disruption in oysters, with significant impacts on offspring.microplastic | reproduction | energy allocation | oyster

Plastic pollution in the environment is increasing at global scale. Microplastics (MP) are derived from degradation of larger plastic items or directly produced in microparticles form (< 5 mm). Plastics, widely used in structures and equipment of pearl farming, are a source of pollution to the detriment of the lagoon ecosystem. To evaluate the impact of MP on the physiology of Pinctada margaritifera, a species of ecological and commercial interests, adult oysters were exposed to polystyrene microbeads (micro-PS of 6 and 10 μm) for 2 months. Three concentrations, 0.25, 2.5, and 25 μg L, and a control were tested. Ingestion and respiration rate and assimilation efficiency were monitored on a metabolic measurement system to determine the individual energy balance (Scope For Growth, SFG). Effects on reproduction were also assessed. The assimilation efficiency decreased significantly according to micro-PS concentration. The SFG was significantly impacted by a dose-dependent decrease from 0.25 μg L ( p < 0.0001), and a negative SFG was measured in oysters exposed to 25 μg L. Gonads may have provided the missing energy to maintain animals' metabolism through the production of metabolites derived from germ cells phagocytosis. This study shows that micro-PS significantly impact the assimilation efficiency and more broadly the energy balance of P. margaritifera, with negative repercussions on reproduction.

This study investigated the potential of RNA interference, which is technically challenging in bivalve mollusc species, to assess gene function in the oyster Crassostrea gigas. We designed dsRNA targeting the oyster vasa‐like gene (Oyvlg), specifically expressed in oyster germ cells. In vivo injection of oyvl‐dsRNA into the gonad provokes a knockdown phenotype corresponding to germ cell underproliferation and prematurely arrested meiosis througout the organ. The most severe phenotype observed is sterile. This knockdown phenotype is associated with a decrease in Oyvlg mRNA level of between 39% and 87%, and a strong reduction in OYVLG protein, to an undetectable level. Therefore, Oyvlg appears to be essential for germ cell development in Crassostrea gigas, particularly for mitotic proliferation and early meiosis. Our results demonstrate for the first time that in vivo RNA interference works efficiently in a bivalve species, opening major perspectives for functional genetic studies.

Blooms of the dinoflagellate Alexandrium spp., known as producers of paralytic shellfish toxins (PSTs), are regularly detected on the French coastline. PSTs accumulate into harvested shellfish species, such as the Pacific oyster Crassostrea gigas, and can cause strong disorders to consumers at high doses. The impacts of Alexandrium minutum on C. gigas have often been attributed to its production of PSTs without testing separately the effects of the bioactive extracellular compounds (BECs) with allelopathic, hemolytic, cytotoxic or ichthyotoxic properties, which can also be produced by these algae. The BECs, still uncharacterized, are excreted within the environment thereby impacting not only phytoplankton, zooplankton but also marine invertebrates and fishes, without implicating any PST. The aim of this work was to compare the effects of three strains of A. minutum producing either only PSTs, only BECs, or both PSTs and BECs, on the oyster C. gigas. Behavioral and physiological responses of oysters exposed during 4 days were monitored and showed contrasted behavioral and physiological responses in oysters supposedly depending on produced bioactive substances. The non-PST extracellular-compound-producing strain primarily strongly modified valve-activity behavior of C. gigas and induced hemocyte mobilization within the gills, whereas the PST-producing strain caused inflammatory responses within the digestive gland and disrupted the daily biological rhythm of valve activity behavior. BECs may therefore have a significant harmful effect on the gills, which is one of the first organ in contact with the extracellular substances released in the water by A. minutum. Conversely, the PSTs impact the digestive gland, where they are released and mainly accumulated, after degradation of algal cells during digestion process of bivalves. This study provides a better understanding of the toxicity of A. minutum on oyster and highlights the significant role of BECs in this toxicity calling for further chemical characterization of these substances.

Summer mortality of the Pacific oyster Crassostrea gigas is the result of a complex interaction between oysters, their environment and their pathogens. The physiological status of an oyster, especially its reproductive status, is suspected to play a significant role in the outcome of this interaction. As genetic variability exists for susceptibility to summer mortality, resistant (R) and susceptible (S) oyster lines were produced using a divergent selection scheme. The present paper reports a histological study on gonad area, which is representative of reproductive effort, in randomly chosen five R and five S oyster lines. The R lines showed a significantly lower gonad area than the S lines (P < 0.001), with an estimated mean difference of 12.5%, whereas, taken together, R and S lines showed a similar distribution of gametogenic stages when sampled. Considering the lines separately, the significant difference in gonad area went up to 24% between R and S lines. The present data confirm and strengthen the negative correlation between reproductive effort and resistance to summer mortality observed in previous studies. Summer mortality of C. gigas in France could, therefore, be partly due to a physiological disorder and metabolic disturbance in oysters associated with their reproductive effort. This does not, however, imply a direct link between the cost of reproduction and mortality because other causal factors, such as pathogenic agents, could be the primary causal factors.

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BackgroundCultured pearls are unique gems produced by living organisms, mainly molluscs of the Pinctada genus, through the biomineralization properties of pearl sac tissue. Improvement of P. margaritifera pearl quality is one of the biggest challenges that Polynesian research has faced to date. To achieve this goal, a better understanding of the complex mechanisms related to nacre and pearl formation is essential and can now be approached through the use of massive parallel sequencing technologies. The aim of this study was to use RNA-seq to compare whole transcriptome expression of pearl sacs that had producing pearls with high and low quality. For this purpose, a comprehensive reference transcriptome of P. margaritifera was built based on multi-tissue sampling (mantle, gonad, whole animal), including different living stages (juvenile, adults) and phenotypes (colour morphotypes, sex).ResultsStrikingly, few genes were found to be up-regulated for high quality pearls (n = 16) compared to the up-regulated genes in low quality pearls (n = 246). Biomineralization genes up-regulated in low quality pearls were specific to prismatic and prism-nacre layers. Alternative splicing was further identified in several key biomineralization genes based on a recent P. margaritifera draft genome.ConclusionThis study lifts the veil on the multi-level regulation of biomineralization genes associated with pearl quality determination.Electronic supplementary materialThe online version of this article (10.1186/s12864-019-5443-5) contains supplementary material, which is available to authorized users.