Toxicity of environmental and polystyrene plastic particles on the bivalve Corbicula fluminea: focus on the molecular responses.

Among aquatic organisms, filter feeders are particularly exposed to the ingestion of microplastics (MPs) and nanoplastics (NPs). The present study investigates the effect of environmental microplastics (ENV MPs) and nanoplastics (ENV NPs) generated from macro-sized plastic debris collected in the Garonne River (France), and polystyrene NPs (PS NPs) on the freshwater bivalve Corbicula fluminea. Organisms were exposed to plastic particles at three concentrations: 0.008, 10, and 100 µg L-1 for 21 days. Gene expression measurements were conducted in gills and visceral mass at 7 and 21 days to assess the effects of plastic particles on different functions. Our results revealed: (i) an up-regulation of genes, mainly involved in endocytosis, oxidative stress, immunity, apoptosis, and neurotoxicity, at 7 days of exposure for almost all environmental plastic particles and at 21 days of exposure for PS NPs in the gills, (ii) PS NPs at the three concentrations tested and ENV MPs at 0.008 µg L-1 induced strong down-regulation of genes involved in detoxication, oxidative stress, immunity, apoptosis, and neurotoxicity at 7 days of exposure in the visceral mass whereas ENV MPs at 10 and 100 µg L-1 and all ENV NPs induced less pronounced effects, (iii) overall, PS NPs and ENV MPs 0.008 µg L-1 did not trigger the same effects as ENV MPs 10 and 100 µg L-1 and all ENV NPs, either in the gills or the visceral mass at 7 and 21 days of exposure. This study highlighted the need to use MPs and NPs sampled in the environment for future studies as their properties induce different effects at the molecular level to living organisms.

Toxicity assessment of microplastics within the Loire River in the clam Corbicula fluminea exposed to environmentally relevant exposure conditions.

The dispersed pollution caused by microplastics (MPs) represents a current and global concern. While the fragmentation of plastic debris into smaller particles occurs in rivers, little MP research is done on freshwater species and is published compared to the marine environment. The Loire River is the longest river in France and is subject to moderate to high anthropic pressure while it represents major societal and economic issues. However, there are not many studies that have been put forward with regards to the effect of environmental MPs (EMPs) on aquatic organisms and no policies have been enacted to monitor the plastic pollution. In this study, freshwater bivalves, Corbicula fluminea, were exposed for 21 days to environmentally relevant concentrations of a mixture of <200 µm MPs generated from plastic litter collected directly along the banks of the Loire River. This mixture was composed of 40% polyethylene (PE), 40% polypropylene (PP), 10% polyethylene terephthalate (PET) and 10% polyvinylchloride (PVC) (mass percentage). Ecotoxicological effects were assessed from the individual to the molecular levels on several endpoints: condition index, filtration efficiency, enzyme activities, lipid peroxidation, energy reserves and gene expression. The ingestion of EMPs caused damages at the biochemical level. Indeed, we reported an increase in catalase activity in gills and digestive mass, a decrease in TBARs in gills, a decrease in acetylcholinesterase activity in the digestive mass, a decrease of glycogen and lipid contents in the whole organisms and a significant induction of the expression of gst, cat, mp, acp genes. The current results suggest therefore that long-term exposure to realistic doses of EMPs causes toxicity towards freshwater benthic biota. The analysis of biomarker activities and the analysis of gene expression are complementary to prevent the effects of a plastic contamination at higher biological levels in aquatic organisms.

Toxicity assessment of environmental MPs and NPs and polystyrene NPs on the bivalve Corbicula fluminea using a multi-marker approach.

Small plastic particles, microplastics (MPs) and nanoplastics (NPs) represent a major threat in aquatic environments. Freshwater organisms are exposed to MPs and NPs, particularly in industrial and urban areas. The present study aimed to compare the toxicity between polystyrene NPs (PS NPs) and environmental microplastics (ENV MPs) and nanoplastics (ENV NPs) generated from macro-sized debris collected in the Garonne River on the freshwater bivalve C. fluminea. The organisms were exposed to the different plastic particles at three environmentally relevant concentrations: 0.008, 10, and 100 µg L-1 for 21 days. The biological responses of organisms were assessed using a multi-biomarker approach from the sub-individual to the individual level. The results demonstrated that: i) ENV NPs triggered more effects on detoxification processes and immune response, confirming that using manufactured NPs for laboratory exposure can lead to misleading conclusions on the risks posed by plastic particles; ii) effects of ENV MPs were less marked than ENV NPs, emphasizing the importance of testing a size continuum of plastic particles from NPs to MPs; iii) some effects were only observed for the low and/or intermediate concentrations tested, underlining the importance of using environmentally relevant concentrations. In light of these results, laboratory studies should be continued by exposing aquatic species to environmental MPs and NPs. The properties of these particles have to be characterized for a better risk assessment of environmental plastic particles.

Aquatic worms: relevant model organisms to investigate pollution of microplastics throughout the freshwater-marine continuum.

Plastic pollution has become a global and emergency concern. Degradation processes of plastic macrowaste, either at the millimetre- and micrometre-size scales (microplastics, MP) or a nanometre one (nanoplastic, NP), is now well documented in all environmental compartments. It is hence necessary to study the environmental dynamic of MNP (micro(nano)plastic) on aquatic macrofauna considering their dispersion in different compartments. In this context, worms, having a large habitat in natural environments (soil, sediment, water) represent a relevant model organism for MNP investigations. In aquatic systems, worms could be used to compare MNP contamination between freshwater and seawater. The aim of this review was to discuss the relevance of using worms as model species for investigating MNP pollution in freshwater, estuarine, and marine systems. In this context, studies conducted in the field and in laboratory, using diverse classes of aquatic worms (polychaete and clitellate, i.e. oligochaete and hirudinea) to assess plastic contamination, were analysed. In addition, the reliability between laboratory exposure conditions and the investigation in the field was discussed. Finally, in a context of plastic use regulation, based on the literature, some recommendations about model species, environmental relevance, and experimental needs related to MNP are given for future studies.

Continuum from microplastics to nanoplastics: effects of size and source on the estuarine bivalve Scrobicularia plana.

Plastic has been largely detected in estuarine environments and represents major concern towards aquatic living organisms. The present study evaluates the impact of microplastics (MPs) and nanoplastics (NPs) under realistic exposure conditions. Scrobicularia plana individuals were exposed to low concentrations (0.008, 10, and 100 µg L-1) of environmental MPs and NPs as well as to standard PS NPs, as a comparison condition. The aim of this study was to understand the ecotoxicological effects of environmental plastic particles on S. plana gills and digestive glands but also to compare the effects of plastic polymers size in order to highlight if the size could induce different toxicity profiles within this model organism, at different levels of biological organization. Results showed a differential induction of detoxification enzymes (CAT, GST), immunity (AcP), DNA damage processes as well as a differential effect on behavior and condition index of animals depending upon the type of plastic, the size, the concentration tested, and the type of organ. This study underlines the necessity of testing (i) plastics collected from the environment as compared to standard ones and (ii) the effect of size using plastics coming from the same batch of macrosized plastics. This study concludes on the future need directions that plastic-based studies must take in order to be able to generate a large quantity of relevant data that could be used for future regulatory needs on the use of plastic.

Effect of size continuum from nanoplastics to microplastics on marine mussel Mytilus edulis: Comparison in vitro/in vivo exposure scenarios.

For several decades, plastic has been a global threat in terms of pollution. Plastic polymers, when introduce in the aquatic environment, are exposed to fragmentation processes into microplastics (MPs) and nanoplastics (NPs) which could potentially interact with living organisms. The objective of this work was to study the effects of plastic particles representative of those found in the environment, on the marine mussels Mytilus edulis, under two exposure scenarios: in vivo and in vitro. Whole mussels or cultured hemocytes were exposed for 24 h to NPs and MPs generated from macro-sized plastics collected in the field, but also to reference NPs, at concentrations found in the environment: 0.08, 10 µg and 100 µg·L-1. Results showed that immune response was only activated when mussels were exposed in vivo. However, cytotoxicity (hemocyte mortality) and genotoxicity (DNA damage) parameters were induced after both types of exposure, but in a dose-dependent manner after in vitro hemocyte exposure to all tested plastic conditions. These results indicate that in vitro approaches could be considered as potential predictors of in vivo exposures.

The need to investigate continuums of plastic particle diversity, brackish environments and trophic transfer to assess the risk of micro and nanoplastics on aquatic organisms.

Plastic particles are ubiquitous in marine and freshwater environments. While many studies have focused on the toxicity of microplastics (MPs) and nanoplastics (NPs) in aquatic environments there is no clear conclusion on their environmental risk, which can be attributed to a lack of standardization of protocols for in situ sampling, laboratory experiments and analyzes. There are also far more studies concerning marine environments than fresh or brackish waters despite their role in the transfer of plastics from continents to oceansWe systematically reviewed the literature for studies: (1) using plastics representative of those found in the environment in laboratory experiments, (2) on the contamination of plastic particles in the continuum between fresh and marine waters, focusing in particular on estuaries and (3) on the continuum of contamination of plastic particles between species through trophic transfer in aquatic environments. We found that the exposure of aquatic organisms in the laboratory to plastic particles collected in the environment are very scarce. Moreover, plastic exposures of estuarine species in the laboratory are generally carried out for a single salinity and a single temperature that do not reflect the fluctuating environmental conditions of estuaries. Finally, the trophic transfer of plastic particles is mainly studied in the laboratory through simple food chains which are not representative of the complexity of the trophic networks observed in the aquatic environment. We pointed out that future studies in the laboratory should include both MPs and NPs sampled in the environment and focus on the precise characterization of the composition and surface of these plastics as well as on their absorbed pollutants, additives or biofilms. Moreover, investigations must be continued concerning the toxicity of plastic particles in brackish water environments such as estuaries and the trophic transfer of plastic particles in complex food chains.

Influence of water temperature and food on the last stages of cultured pearl mineralization from the black-lip pearl oyster Pinctada margaritifera.

Environmental parameters, such as food level and water temperature, have been shown to be major factors influencing pearl oyster shell growth and molecular mechanisms involved in this biomineralization process. The present study investigates the effect of food level (i.e., microalgal concentration) and water temperature, in laboratory controlled conditions, on the last stages of pearl mineralization in order to assess their impact on pearl quality. To this end, grafted pearl oysters were fed at different levels of food and subjected to different water temperatures one month prior to harvest to evaluate the effect of these factors on 1) pearl and shell deposition rate, 2) expression of genes involved in biomineralization in pearl sacs, 3) nacre ultrastructure (tablet thickness and number of tablets deposited per day) and 4) pearl quality traits. Our results revealed that high water temperature stimulates both shell and pearl deposition rates. However, low water temperature led to thinner nacre tablets, a lower number of tablets deposited per day and impacted pearl quality with better luster and fewer defects. Conversely, the two tested food level had no significant effects on shell and pearl growth, pearl nacre ultrastructure or pearl quality. However, one gene, Aspein, was significantly downregulated in high food levels. These results will be helpful for the pearl industry. A wise strategy to increase pearl quality would be to rear pearl oysters at a high water temperature to increase pearl growth and consequently pearl size; and to harvest pearls after a period of low water temperature to enhance luster and to reduce the number of defects.