Pearl Farming Micro-Nanoplastics Affect Oyster Physiology and Pearl Quality.

Pearl farming is crucial for the economy of French Polynesia. However, rearing structures contribute significantly to plastic waste, and the widespread contamination of pearl farming lagoons by microplastics has raised concerns about risks to the pearl industry. This study aimed to evaluate the effects of micro-nanoplastics (MNPs, 0.4-200 µm) on the pearl oyster (Pinctada margaritifera) over a 5-month pearl production cycle by closely mimicking ecological scenarios. MNPs were produced from weathered plastic pearl farming gear and tested at environmentally relevant concentrations (0.025 and 1 µg L-1) to decipher biological and functional responses through integrative approaches. The significant findings highlighted the impacts of MNPs on oyster physiology and pearl quality, even at remarkably low concentrations. Exposure to MNPs induced changes in energy metabolism, predominantly driven by reduced assimilation efficiency of microalgae, leading to an alteration in gene expression patterns. A distinct gene expression module exhibited a strong correlation with physiological parameters affected by MNP conditions, identifying key genes as potential environmental indicators of nutritional-MNP stress in cultured oysters. The alteration in pearl biomineralization, evidenced by thinner aragonite crystals and the presence of abnormal biomineral concretions, known as keshi pearls, raises concerns about the potential long-term impact on the Polynesian pearl industry.

Cryogrinding and sieving techniques as challenges towards producing controlled size range microplastics for relevant ecotoxicological tests.

The impact of microplastics (MP) has attracted much attention from the scientific community and many laboratory assessments have been made of their effects on aquatic organisms. To produce MP from real environmental plastic waste, which would enable more realistic experiments, we used plastic pearl farming equipment from French Polynesian lagoons. Here, the pearl oyster Pinctada margaritifera could encounter MP coming from their breakdown in its surrounding environment. We tested an established method based on mechanical cryogenic grinding and liquid sieving. Our desired size range was 20-60 µm, corresponding to the optimal particle size ingested by P. margaritifera. The protocol was effective, generating MP particles of 20-60 µm (∼17,000-28,000 MP µg-1), but also produced too many smaller particles. The peak in the desired size range was thus flattened by the many small particles <3 µm (∼82,000-333,000 MP µg-1; 53-70% of total analysed particles), visible at the limit of Coulter counter analysis (cut-off point: 2 µm). Laser diffraction analysis (cut-off point: 0.4 µm) provided greater detail, showing that ∼80-90% of the total analysed particles were <1 µm. Diverging particle size distributions between those expected based on sieving range and those really observed, highlight the need to perform fine-scaled particle size distribution analyses to avoid underestimating the number of small micro- and nanoplastics (MNP) and to obtain an exact estimation of the fractions produced. Size and microstructure characterization by scanning electron microscopy suggested spontaneous particle self-assembly into crystal superstructures, which is the supposed cause of the divergence we observed. Overall, our results emphasize that particle self-assembly is a technical hurdle requiring further work and highlight the specific need to finely characterize the size distribution of MNP used in ecotoxicological experiments to avoid overestimating effects.

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.

Microplastics contamination in pearl-farming lagoons of French Polynesia.

Pearl-farming is the second most important source of income in French Polynesia. However, tropical lagoons are fragile ecosystems with regard to anthropogenic pressures like plastic pollution, which threaten marine life and the pearl oyster-related economy. Here, we investigated the spatial distribution of microplastics (MP) and concentrations in surface water (SW), water column (WC) and cultivated pearl oyster (PO) from three pearl-farming atolls with low population and tourism. Microplastics were categorized by their size class, shape, colour and polymer type identified using FTIR spectroscopy. Widespread MP contamination was observed in every study site (SW, 0.2-8.4 MP m-3; WC, 14.0-716.2 MP m-3; PO, 2.1-125.0 MP g-1 dry weight), with high contamination in the WC highlighting the need to study the vertical distribution of MP, especially as this compartment where PO are reared. A large presence of small (< 200 µm) and fragment-shaped (> 70%) MP suggests that they result from the breakdown of larger plastic debris. The most abundant polymer type was polyethylene in SW (34-39%), WC (24-32%), while in PO, polypropylene (14-20%) and polyethylene were more evenly distributed (9-21%). The most common MP identified as black-grey polyethylene and polypropylene matches the polymer and colour of ropes and collectors questioning a pearl-farming origin.

Effects of elevated temperature and pCO2 on the respiration, biomineralization and photophysiology of the giant clam Tridacna maxima.

Many reef organisms, such as the giant clams, are confronted with global change effects. Abnormally high seawater temperatures can lead to mass bleaching events and subsequent mortality, while ocean acidification may impact biomineralization processes. Despite its strong ecological and socio-economic importance, its responses to these threats still need to be explored. We investigated physiological responses of 4-year-old Tridacna maxima to realistic levels of temperature (+1.5°C) and partial pressure of carbon dioxide (pCO2) (+800 µatm of CO2) predicted for 2100 in French Polynesian lagoons during the warmer season. During a 65-day crossed-factorial experiment, individuals were exposed to two temperatures (29.2°C, 30.7°C) and two pCO2 (430 µatm, 1212 µatm) conditions. The impact of each environmental parameter and their potential synergetic effect were evaluated based on respiration, biomineralization and photophysiology. Kinetics of thermal and/or acidification stress were evaluated by performing measurements at different times of exposure (29, 41, 53, 65 days). At 30.7°C, the holobiont O2 production, symbiont photosynthetic yield and density were negatively impacted. High pCO2 had a significant negative effect on shell growth rate, symbiont photosynthetic yield and density. No significant differences of the shell microstructure were observed between control and experimental conditions in the first 29 days; however, modifications (i.e. less-cohesive lamellae) appeared from 41 days in all temperature and pCO2 conditions. No significant synergetic effect was found. Present thermal conditions (29.2°C) appeared to be sufficiently stressful to induce a host acclimatization response. All these observations indicate that temperature and pCO2 are both forcing variables affecting T. maxima's physiology and jeopardize its survival under environmental conditions predicted for the end of this century.

Microplastics induce dose-specific transcriptomic disruptions in energy metabolism and immunity of the pearl oyster Pinctada margaritifera.

A combined approach integrating bioenergetics and major biological activities is essential to properly understand the impact of microplastics (MP) on marine organisms. Following experimental exposure of polystyrene microbeads (micro-PS of 6 and 10 µm) at 0.25, 2.5, and 25 µg L-1, which demonstrated a dose-dependent decrease of energy balance in the pearl oyster Pinctada margaritifera, a transcriptomic study was conducted on mantle tissue. Transcriptomic data helped us to decipher the molecular mechanisms involved in P. margaritifera responses to micro-PS and search more broadly for effects on energetically expensive maintenance functions. Genes related to the detoxification process were impacted by long-term micro-PS exposure through a decrease in antioxidant response functioning, most likely leading to oxidative stress and damage, especially at higher micro-PS doses. The immune response was also found to be dose-specific, with a stress-related activity stimulated by the lowest dose present after a 2-month exposure period. This stress response was not observed following exposure to higher doses, reflecting an energy-limited capacity of pearl oysters to cope with prolonged stress and a dramatic shift to adjust to pessimum conditions, mostly limited and hampered by a lowered energetic budget. This preliminary experiment lays the foundation for exploring pathways and gene expression in P. margaritifera, and marine mollusks in general, under MP exposure. We also propose a conceptual framework to properly assess realistic MP effects on organisms and population resilience in future investigations.

Toxic effects of leachates from plastic pearl-farming gear on embryo-larval development in the pearl oyster Pinctada margaritifera.

Pearl-farming leads to significant plastic pollution in French Polynesia (FP) as the end of life of most farming gear is currently poorly managed. Plastic debris released into the aquatic environment accumulates, with potentially detrimental effects on the lagoon ecosystem and pearl oyster Pinctada margaritifera, a species of ecological, commercial and social value. Here, we tested the effects of leachates from new (N) and aged (A) plastic pearl-farming gear (spat collector and synthetic rope) obtained after 24 h and 120 h incubation, on the embryo-larval development of the pearl oyster using an in-vitro assay. Embryos were exposed for 24 h and 48 h to a negative control (0) and the leachate from 0.1, 1, 10 and 100 g of plastic. L-1. After 24 h exposure to leachate at 100 g.L-1, effects were observed on embryo development (-38% to -60% of formed larvae) and mortality (+72% to +82%). Chemical analyses of plastic gear indicated the presence of 26 compounds, consisting of organic contaminants (PAHs) and additives (mainly phthalates). Screening of leachates demonstrated that these compounds leach into the surrounding seawater with an additional detection of pesticides. Higher levels of phthalates were measured in leachates obtained from new (6.7-9.1 µg.L-1) than from aged (0.4-0.5 µg.L-1) plastics, which could be part of the explanation of the clear difference in toxicity observed after 48 h exposure at lower concentrations (0.1-10 g.L-1), associated with mortality ranging from 26 to 86% and 17-28%, respectively. Overall, this study suggests that plastic gear used in the pearl-farming industry releases significant amounts of hazardous chemicals over their lifetime, which may affect pearl oyster development that call for in-situ exploration.

Influence of temperature and pearl rotation on biomineralization in the pearl oyster, Pinctada margaritifera.

The objective of this study was to observe the impact of temperature on pearl formation using an integrative approach describing the rotation of the pearls, the rate of nacre deposition, the thickness of the aragonite tablets and the biomineralizing potential of the pearl sac tissue though the expression level of some key genes. Fifty pearl oysters were grafted with magnetized nuclei to allow the rotation of the pearls to be described. Four months later, 32 of these pearl oysters were exposed to four temperatures (22, 26, 30 and 34°C) for 2 weeks. Results showed that the rotation speed differed according to the movement direction: pearls with axial movement had a significantly higher rotation speed than those with random movement. Pearl growth rate was influenced by temperature, with a maximum between 26 and 30°C but almost no growth at 34°C. Lastly, among the nine genes implicated in the biomineralization process, only Pmarg-Pif177 expression was significantly modified by temperature. These results showed that the rotation speed of the pearls was not linked to pearl growth or to the expression profiles of biomineralizing genes targeted in this study. On the basis of our results, we consider that pearl rotation is a more complex process than formerly thought. Mechanisms involved could include a strong environmental forcing in immediate proximity to the pearl. Another implication of our findings is that, in the context of ocean warming, pearl growth and quality can be expected to decrease in pearl oysters exposed to temperatures above 30°C.

Low energy cost for cultured pearl formation in grafted chimeric Pinctada margaritifera.

The pearl oyster is one of the rare animal models that support two distinct genomes, through the surgical graft process operated for culture pearl production. This grafted organism is assimilated to a chimera whose physiological functioning remains poorly known. The question of the energy expenditure comparison between chimera and non-chimera animals arises. To answer this question, grafted and non-grafted pearl oysters were evaluated for their energetic needs by the indirect calorimetry method. This method made it possible to measure the energy expenditure based on the respiration rate (RR) measurement, reflecting the basal metabolism. The results showed that the RR values for grafted and non-grafted pearl oysters were not significantly different (p < 0.05). The estimated cost of pearl calcification including CaCO3 and proteins synthesis was 0.237 ± 0.064 J h-1, representing 0.64% of the total energy expenditure of grafted pearl oysters. This study made it possible, for the first time, to see the energy cost of cultured pearl formation in P. margaritifera and the little impact in the energetic metabolism of the chimera organism.

Microplastics Affect Energy Balance and Gametogenesis in the Pearl Oyster Pinctada margaritifera.

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-1, 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-1 ( p < 0.0001), and a negative SFG was measured in oysters exposed to 25 µg L-1. 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.

Phenome of pearl quality traits in the mollusc transplant model Pinctada margaritifera.

The bivalve Pinctada margaritifera exhibits three main transplant phenotypes derived from the donor (from which a mantle graft tissue, the saibo, is excised), the recipient (into which the saibo is implanted with a nucleus, leading to the formation of a pearl sac "chimera") and the cultured pearls themselves. This first phenome study on the species derived from a large experimental graft. Transplant phenotype was assessed at three scales: 1) macro, pearl size, colour, grade, 2) micro, pearl surface microstructure, and 3) molecular, biomineralisation gene expression level in saibo and pearl sac tissues. From donor to pearl, the phenome revealed fine variations of quality traits dependent on the position on the mantle where the saibo was cut, whose variation could overlap with inter-individual donor phenotype differences. A single donor phenotype could therefore produce multiple pearl phenotypes at the scale of the saibo position, mirroring its original activity at the mantle position level and the colour and shape of the shell. This phenome study provides essential information on phenotypic trait architecture enabling us to explore and explain the main biological functions and pave the way for a phenomic project on P. margaritifera that could benefit the pearl industry.

Temperature and food influence shell growth and mantle gene expression of shell matrix proteins in the pearl oyster Pinctada margaritifera.

In this study, we analyzed the combined effect of microalgal concentration and temperature on the shell growth of the bivalve Pinctada margaritifera and the molecular mechanisms underlying this biomineralization process. Shell growth was measured after two months of rearing in experimental conditions, using calcein staining of the calcified structures. Molecular mechanisms were studied though the expression of 11 genes encoding proteins implicated in the biomineralization process, which was assessed in the mantle. We showed that shell growth is influenced by both microalgal concentration and temperature, and that these environmental factors also regulate the expression of most of the genes studied. Gene expression measurement of shell matrix protein thereby appears to be an appropriate indicator for the evaluation of the biomineralization activity in the pearl oyster P. margaritifera under varying environmental conditions. This study provides valuable information on the molecular mechanisms of mollusk shell growth and its environmental control.