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Plastic products contain complex mixtures of chemical compounds that are incorporated into polymers to improve material properties. Besides the intentional chemical additives, other compounds including residual monomers and non-intentionnaly added substances (NIAS) as well as sorbed pollutants are usually also present in aged plastic. Since most of these substances are only loosely bound to the polymer via non-covalently interactions, i.e., van der Waals forces, they may leach to the surrounding environment. Although there is increasing knowledge about toxicity of weathered plastic to aquatic organisms, only little is known about how plastic associated chemicals affect human health. Seafood consumption is one of the routes of human exposure to microplastics. The aim of this study was to evaluate the ability of naturally aged plastic associated chemicals to induce harmful effects to human health via the consumption of MP-contaminated seafood. Human colorectal adenocarcinoma Caco-2 and human hepatocyte carcinoma HepG2 cells were selected as model of the colon and liver cells respectively. They are known for their high capacity to metabolize organic contaminants. Both cell lines were exposed to DMSO extracts of different plastics to investigate the effects of chemicals on cell viability, oxidative stress induction and genotoxicity. In addition, the estrogenic effects of DMSO-extracts were evaluated using an estrogen-dependent reporter gene assay in T47D-Kbluc human breast cancer cells. Chemical profiles of the DMSO extracts were polymer-dependent, with polyvinyl chloride (PVC) highly contaminated with metals while polypropylene (PP) contained the lowest concentration of metals. Organic pollutants, including polycyclic aromatic hydrocarbons, were mainly found in PVC, high density polyethylene (HDPE) and PP extracts, whereas other extracted plastics had less (PP) to no organic contamination (polyethylene terephthalate PET). PVC was the most toxic plastic inducing cytotoxicity for both cell lines. DNA damage was observed for Caco-2 cells exposure to HDPE, PVC and nylon. Reactive oxygen species were induced only with nylon extracts in intestinal cells. No toxicity was observed for PP and PET and none of the tested plastics had any estrogenic effect. Our results demonstrate that some environmental aged plastic material released a variety of known and unknown chemical compounds some of which are toxic in vitro and contribute to the knowledge on adverse human health effects of plastics., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Benedicte MORIN reports financial support was provided by SIBA. Benedicte MORIN reports financial support was provided by Nouvelle-Aquitaine Regional Council. Benedicte MORIN reports financial support was provided by French Biodiversity Office. Benedicte MORIN reports financial support was provided by French water agency Adour Garonne. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025 The Authors. Published by Elsevier Inc. All rights reserved.)

In the context of increasing pollution pressure on aquatic ecosystems, it is essential to improve our knowledge of habitat quality and its suitability for organisms. It is particularly relevant to better integrate early life stages of fish into pollution biomonitoring programs, as they are reliable indicators of ecosystem integrity and because of their high sensitivity to pollutants. To avoid the influence of environmental parameters on their development, a lab-on-field approach, called the ex-situ exposure method, was developed. Aquatic organisms were exposed to a continuous flux of water under semi-controlled temperature, oxygen, and photoperiod conditions to avoid the influence of these confounding factors when interpreting the results. To investigate the potential role of water contamination, this active biomonitoring method was applied to the Garonne River (Southwest France), where migratory fish populations have declined. Two model species from different taxa were used: embryos of the Japanese medaka (Oryzias latipes) and adults of the crustacean Gammarus fossarum. The results showed a significant impact of water quality on embryo mortality and early hatching in two separate experiments on Japanese medaka. In addition, an induction of feeding rate was observed in exposed gammarids, but no impact on their embryo survival, suggesting differences in sensitivity between the two species selected. Chemical and biological analyses did not identify trace metals, pesticides, or microorganisms as potential sources of toxicity in medaka embryos or G. fossarum. These results raise concerns about the quality of the water in the Garonne River and its toxicity to aquatic organisms., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Sarah Bancel reports financial support was provided by Regional Water Agency Adour-Garonne. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Conventional plastics have become a major environmental concern due to their persistence and accumulation in marine ecosystems. The development of potential degradable polymers (PBP), such as polyhydroxyalkanoates (PHAs) and polylactic acid (PLA), has gained attention as an alternative to mitigate plastic pollution, since they have the potential to biodegrade under certain conditions, and their production is increasing as replacement of conventional polyolefins. This study aimed to assess and compare the toxicity of leachates of pre-compounding PBP (PLA and the PHA, polyhydroxybutyrate-covalerate (PHBv)) and polypropylene (PP) on five marine planktonic species. A battery of standard bioassays using bacteria, microalgae, sea urchin embryos, mussel embryos and copepod nauplii was conducted to assess the toxicity of leachates from those polymers. Additionally, the presence of chemical additives in the leachates was also verified through GC-MS and LC-HRMS analysis. Results showed that PHBv leachates exhibited higher toxicity compared to other polymers, with the microalgae Rhodomonas salina, being the most sensitive species to the tested leachates. On the other hand, PP and PLA generally displayed minimal to no toxicity in the studied species. Estimated species sensitivity distribution curves (SSD) show that PHBv leachates can be 10 times more hazardous to marine plankton than PP or PLA leachates, as demonstrated by the calculated Hazardous Concentration for 5 % of species (HC 5 ). Qualitative chemical analysis supports the toxicological results, with 80 % of compounds being identified in PHBv leachates of which 2,4,6-trichlorophenol is worth mentioning due to the deleterious effects to aquatic biota described in literature. These findings underscore the fact that whereas environmental persistence can be targeted using PBP, the issue of chemical safety remains unsolved by some alternatives, such as PHBv. Gaining a comprehensive understanding of the toxicity profiles of PBP materials through a priori toxicological risk assessment is vital for their responsible application as alternatives to conventional plastics., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Water pollution is a one of the most contributors to aquatic biodiversity decline. Consequently, ecological risk assessment methods have been developed to investigate the effects of existing stresses on the environment, including the toxic effects of chemicals. One of the existing approaches to quantify toxic risks is called "Potentially Affected Fraction of species" (PAF), which estimates the potential loss of species within a group of species studied. In this study, the PAF method was applied to the Garonne catchment (southwest France) due to the limited information available on the involvement of water pollution in the decline of diadromous fish populations. This approach was used to quantify the potential toxic risk associated with chemical contamination of water for fish species. The objectives were to quantify this risk (1) in the Garonne and Dordogne rivers and (2) in the spawning grounds of two endangered anadromous fish species: the allis shad and the European sturgeon during the development period of their early life stages. Environmental pollution data was provided for 21 sites within the Garonne catchment between 2007 and 2022, and toxicity data was obtained specifically from freshwater toxicity tests on fish species. Then, for each site and each year, the potential toxic risk for a single substance (ssPAF) and for a mixture of substances (msPAF) was calculated and classified as high (>5 %), moderate (>1 % and < 5 %) or low (<1 %). Potential toxic risks were mostly moderate and mainly associated with: metals > other industrial pollutants and hygiene and care products > agrochemicals. In summary, this study highlights the probable involvement of water contamination on the decline, fate and restoration of diadromous fish populations in the Garonne catchment, focusing notably on the toxic effects on early life stages, a previously understudied topic., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Assessment of microplastic (MP) contamination is still needed to evaluate this threat correctly and tackle this issue. Here, MP contamination was assessed for a meso-tidal lagoon of the Atlantic coast (Arcachon Bay, France). Sea surface, water column, intertidal sediments and wild oysters were sampled. Five different stations were studied to assess the spatial distribution of the contamination. Two were outside of the bay and three were inside the bay (from the inlet to the back). A distinction was made between all anthropogenic particles (AP, i.e. visually sorted) and MP (i.e. plastic polymer confirmed by ATR-FTIR spectroscopy). The length of particles recovered in this study ranged between 17 μm and 5 mm. Concentration and composition in sea surface and water column samples showed spatial variations while sediment and oyster samples did not. At outside stations, the sea surface and the water column presented a blended composition regarding shapes and polymers and low to high concentrations (e.g. 0.16 ± 0.08 MP.m -3 and 561.7 ± 68.5 MP.m -3 , respectively for sea surface and water column), which can be due to coastal processes and nearby input sources. The inlet station displayed a well-marked pattern only at the sea surface. High AP and MP concentrations were recorded, and fragments along with polyethylene overwhelmed (respectively 76.0 % and 73.2 %). Higher surface currents could explain this pattern. At the bay back, AP and MP concentrations were lower and fibers were mainly recorded. Weaker hydrodynamics in this area was suspected to drive this contamination profile. Overall, fragments and buoyant particles were mainly detected at the sea surface while fibers and negatively buoyant particles prevailed in other compartments. Most of the studied samples presented an important contribution of fiber-shaped particles (from 31.5 % to 94.2 %). Finally, contamination was ubiquitous as AP and MP were found at all stations in all sample types., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The chemical components of plastic wastes have made their disposal a major economic, social, and environmental problem worldwide. This study evaluated the acute toxicity and genotoxicity of marine plastic debris on the beaches of Concepción Bay, Central Chile, taken during three periods (spring, summer, and winter). An integrated approach was used, including chemical and toxicological data, using the Microtox® test with Vibrio fischeri and SOS chromotest with Escherichia coli and concentrations of polychlorinated biphenyls (PCBs), Organochlorine Pesticides (OCPs) and polybrominated diphenyl ethers (PBDEs). The results presented here exclusively include the novel data obtained from the winter campaign, revealing high concentrations of PBDEs (238 ± 521 ng g -1 ). In addition, the genotoxicity and acute toxicity tests were sensitive for most of the samples studied. This investigation is the first attempt to analyse the toxicity of plastic debris in coastal areas along the Chilean coast., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Carbaryl and fenitrothion are two insecticides sharing a common mode of action, the inhibition of the acetylcholinesterase (AChE) activity. Their use is now regulated or banned in different countries, and the environmental levels of both compounds in aquatic ecosystems have decreased to the range of pg/L to ng/L. As these concentrations are below the non-observed-adverse-effect-concentrations (NOAEC) for AChE inhibition reported for both compounds in aquatic organisms, there is a general agreement that the current levels of these two chemicals are safe for aquatic organisms. In this study we have exposed zebrafish, Japanese medaka and Daphnia magna to concentrations of carbaryl and fenitrothion under their NOAECs for 24-h, and the effects on heart rate (HR), basal locomotor activity (BLA), visual motor response (VMR), startle response (SR) and its habituation have been evaluated. Both pesticides increased the HR in the three selected model organisms, although the intensity of this effect was chemical-, concentration- and organism-dependent. The exposure to both pesticides also led to a decrease in BLA and an increase in VMR in all three species, although this effect was only significant in zebrafish larvae. For SR and its habituation, the response profile was more species- and concentration-specific. The results presented in this manuscript demonstrate that concentrations of carbaryl and fenitrothion well below their respective NOAECs induce tachycardia and the impairment of ecologically relevant behaviors in phylogenetically distinct aquatic model organisms, both vertebrates and invertebrates, emphasizing the need to include this range of concentrations in the environmental risk assessment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Behavioral parameters are increasingly considered sensitive and early bioindicators of toxicity in aquatic organisms. A video-tracking tool was specifically developed to monitor the swimming behaviour of D-larvae of the Mediterranean mussel, Mytilus galloprovincialis, in controlled laboratory conditions. Both maximum and average swimming speeds and trajectories were recorded. We then investigated the impact of copper and silver with or without a moderate rise of temperature on swimming behavior and acetylcholinesterase (AChE) activity of mussel D-larvae and the possible mechanistic link between both biological responses. Our results showed that copper and/or silver exposure, as well as temperature increase, disrupts the swimming behavior of mussel larvae which could compromise their dispersal and survival. In addition, the combined effect of temperature and metals significantly (p < 0.05) increased AChE activity in mussel larvae. Pearson's correlation analysis was performed and results showed that the AChE activity is positively correlated with maximum speeds (r = 0.71, p < 0.01). This study demonstrates the value of behavioral analyzes of aquatic invertebrates as a sensitive and integrate marker of the effects of stressors., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

In the past few decades, glyphosate became the most used herbicide substance worldwide. As a result, the substance is ubiquitous in surface waters. Concerns have been raised about its ecotoxicological impact, but little is known about its generational toxicity. In this study, we investigate the impact of an environmentally relevant concentration of glyphosate and its co-formulants on an F2 generation issued from exposed generations F0 and F1. Trans, inter and multigenerational toxicity of 1 μgL -1 of the active substance was evaluated on early stages of development and juvenile rainbow trout (Oncorhynchus mykiss) using different molecular, biochemical, immuno-hematologic, and biometric parameters, behavior analysis, and a viral challenge. Reproductive parameters of generation F1 were not affected. However, developmental toxicity in generation F2 due to glyphosate alone or co-formulated was observed with head size changes (e.g. head surface up to +10%), and metabolic disruptions (e.g. 35% reduction in cytochrome-c-oxidase). Moreover, larvae exposed transgenerationally to Viaglif and intergenerationally to glyphosate and Roundup presented a reduced response to light, potentially indicating altered escape behavior. Overall methylation was, however, not altered and further experiments using gene-specific DNA metylation analyses are required. After several months, biochemical parameters measured in juvenile fish were no longer impacted, only intergenerational exposure to glyphosate drastically increased the susceptibility of rainbow trout to hematopoietic necrosis virus. This result might be due to a lower antibody response in exposed fish. In conclusion, our results show that generational exposure to glyphosate induces developmental toxicity and increases viral susceptibility. Co-formulants present in glyphosate-based herbicides can modulate the toxicity of the active substance. Further investigations are required to study the specific mechanisms of transmission but our results suggest that both non-genetic mechanisms and exposure during germinal stage could be involved., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The last remaining population of European sturgeon (Acipenser sturio) lives in the Gironde-Garonne-Dordogne (France) catchment (GGD). Captive young individuals are released into the GGD hydrosystem each year, as part of a restocking programme. This study aims to assess the health status of juveniles A. sturio to current conditions in the GGD hydrosystem, to evaluate their capacity to survive and grow in a moderately anthropized ecosystems. 3-month-old farmed sturgeons were exposed for one month in experimental conditions that mimic the environmental conditions in the Garonne and Dordogne rivers, followed by five months of depuration. After one month of exposure, fish exposed to Dordogne and Garonne waters bioaccumulated higher levels of metals and persistent organic pollutants, displayed a reduced hepato-somatic index, and had depleted levels of lipids and glycogen content in their liver, when compared with the Reference group. However, metabolic and swimming performance, as well as the costs of swimming were not impaired. After the 5 months depuration, a significant decrease of K was observed for all exposure conditions. HSI also decreased with time. The overall health status and adaptive capacity of juvenile A. sturio appeared to be maintained over the experimental 6 months' period. Juveniles of A. sturio seem to have the adaptive capacity to survive and grow in the GGD hydrosystem, after being released as part of a restocking programme., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Coastal environments are a predominant ultimate destination of marine debris, becoming a key focus of studies assessing microplastic (MP) contamination. Here, we described the visible fraction of MP (from 0.5 to 5 mm) that washed up during the high tide at different sites of a semi-enclosed mesotidal bay and investigated the main abiotic factors driving MP beaching. Three contrasted beaches of the Arcachon Bay (SW France) were monitored on a monthly basis during 2019. Samplings were made along a 100 m longitudinal transect at the high-water strandline (4 quadrats of 0.25m 2 ) and at an intermediate tidal range. Each sampled particle was characterized by morphometric data (e.g. size, shape, color, roughness) and polymer identification was performed by ATR-FTIR technique. Results show that MP concentration was higher on the beach located at the mouth of the bay (36.0 ± 39.2 MP.m -2 ) than at the back and the outside of the bay (respectively 2.7 ± 4.4 and 1.7 ± 2.4 MP.m -2 ). This may be related to the strong currents at the entry of the embayment and the beach orientation, exposed to predominant winds. Beached MP were mainly pre-production pellets and fragments as they represented respectively 49% and 39% of all analyzed shapes. Polymers with low density were particularly abundant. Polyethylene represented 69% of all the particles while polypropylene accounted for 17% and polystyrene for 10%. We also observed that MP were mostly washed up when wind, waves and river flow were more intense. Analysis suggest that wind direction and speed are key factors influencing beaching as strong onshore wind enhance this process., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Plastic is currently used in aquaculture as a material for settlement and magnification of oyster spats. Plastic weathering and fragmentation under natural conditions can lead to the production of micro and nanoparticles and additive leakage, with potential toxic effects on marine life. This study investigates the effects of the exposure to microplastic (MPs) cocktail derived from aged aquaculture material on oyster pediveliger larvae (Crassostrea gigas). The cocktail was made of high-density polyethylene (HDPE), polypropylene (PP) and polyvinyl chloride (PVC). The concentrations tested were 0, 0.1, and 10 mg MP·L -1 . During the 7-day fixation phase, pediveliger larvae (17 days) were exposed to the MP cocktail in laboratory-controlled conditions. After exposure, the success of settlement was significantly lower for larvae exposed to 10 mg MP·L -1 (49 ± 0.9%) compared to control ones (61.8 ± 1.6%). No malformations or metamorphosis abnormalities were observed. Growth of pediveliger and spat stages was monitored up to 11 months. During the first twenty-eight days of development, spat growth was significantly lower for the two MPs exposure conditions (0.1 and 10 mg MP·L -1 ; respectively -51.8% and -44.4%) compared to control group. Subsequently, the previously exposed oysters grew faster than the control condition, resulting in a significantly greater growth (0.1 and 10 mg MP·L -1 : +18.3% and +19.7%) than the control group at the end of follow-up. The nearly one-year follow-up highlighted the potential effects of MPs from aquaculture on larvae and spat of C. gigas., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Toxicity of polyethylene (PE) and polyvinyl chloride (PVC) microplastics (MPs), either virgin or spiked with chemicals, was evaluated in two short-lived fish using a freshwater species, zebrafish, and a marine species, marine medaka. Exposures were performed through diet using environmentally relevant concentrations of MPs over 4 months. No modification of classical biomarkers, lipid peroxidation, genotoxicity or F0 behaviour was observed. A significant decrease in growth was reported after at least two months of exposure. This decrease was similar between species, independent from the type of MPs polymer and the presence or not of spiked chemicals, but was much stronger in females. The reproduction was evaluated and it revealed a significant decrease in the reproductive output for both species and in far more serious numbers in medaka. PVC appeared more reprotoxic than PE as were MPs spiked with PFOS and benzophenone-3 compared to MPs spiked with benzo[a]pyrene. Further, PVC-benzophenone-3 produced behavioural disruption in offspring larvae. These results obtained with two species representing different aquatic environments suggest that microplastics exert toxic effects, slightly different according to polymers and the presence or not of sorbed chemicals, which may lead in all cases to serious ecological disruptions., (Copyright © 2021 Elsevier B.V. All rights reserved.)

This study aims to assess the toxic effects and the potential risk of widely used agricultural pesticides on the development (malformations and developmental arrest), growth and swimming activity of oyster D-larvae (Crassostrea gigas). Freshly fertilized oyster embryos were exposed for 24 h at 24 °C to different concentrations (0, 0.01, 0.1, 1 and 10 μg.L -1 ) of six different pesticides: Glyphosate and its commercial solution (Roundup), Isoproturon, Nicosulfuron, Chlortoluron and Boscalid. The six pesticides tested induced a significant increase in larval malformations and developmental arrests. All pesticides except Glyphosate and Isoproturon affected larval growth. Roundup, Nicosulfuron, Chlortoluron and Boscalid also affected the swimming behaviour of the D-larvae, with a significant decrease recorded in their maximum swimming speed. Comparison of the LOEC (Lowest-Observed-Effect Concentration) of each compound led to the following toxicity classification: Boscalid > Chlortoluron = Nicosulfuron > Glyphosate > Roundup > Isoproturon, with respectively LOEC of 0.0028; 0.015; 0.017; 0.11; 0.3 and 0.78 μg.L -1 . By comparison of the maximum concentrations in the Pertuis Charentais (South West, France) and LOEC of each pesticide, the following risk scale was obtained: Chlortoluron > Boscalid > Glyphosate > Roundup > Nicosulfuron > Isoproturon. Our results revealed that Chlortoluron, Boscalid and to a lesser extent Glyphosate represent a potential threat to early life stages of oyster living in the Pertuis Charentais marine area., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Worsened state of oysters in French Arcachon Bay, demand an investigation of possible causes. This study evaluated the effects of an environmentally relevant mixture of five common pesticides on the early-life stages of the Pacific oyster (Magallana gigas). Laboratory assays with artificial mixture and in situ transplantation were complementarily used to investigate a series of sublethal endpoints. The laboratory exposure revealed developmental toxicity at 0.32 μg/L, which corresponds to mixture concentrations in Arcachon Bay. Downregulation of some gene transcriptions was observed at environmental level. No difference in larvae development was revealed among the three sites in Arcachon Bay. This study was the first to evaluate locomotion of oyster larvae exposed in situ. Suspected poor water quality in the inner part of Arcachon Bay was reflected by impairment at the molecular level. In conclusion, current concentrations of the tested pesticides in Arcachon Bay hinder larval development and affect several biological functions., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Coastal areas are final recipients of various contaminants including pesticides. The effects of pesticides on non-target organisms are often unclear, especially at environmentally relevant concentrations. This study investigated the impacts of insecticide imidacloprid (IMI) and fungicide propiconazole (PRO), some of the most detected pesticides in the Arcachon Bay in France. This work also included the research of propiconazole nanoformulation (nanoPRO). The effects were assessed studying the development of the early life stages of the Pacific oyster (Magallana gigas). Oyster embryos were exposed for 24, 30, and 42 h (depending on the endpoint) at 24 °C to environmentally relevant concentrations of the two pesticides as well as to nanoPRO. The research focused on sublethal endpoints such as the presence of developmental malformations, alterations of locomotion patterns, or changes in the gene expression levels. No developmental abnormalities were observed after exposure to environmental concentrations detected in the Arcachon Bay in recent years (maximal detected concentration of IMI and PRO were 174 ng/L and 29 ng/L, respectively). EC 50 of PRO and nanoPRO were comparable, 2.93 ± 1.35 and 2.26 ± 1.36 mg/L, while EC 50 of IMI exceeded 200 mg/L. IMI did not affect larval behavior. PRO affected larval movement trajectory and decreased average larvae swimming speed (2 μg/L), while nanoPRO increased the maximal larvae swimming speed (0.02 μg/L). PRO upregulated especially genes linked to reactive oxygen species (ROS) production and detoxification. NanoPRO effects on gene expression were less pronounced - half of the genes were altered in comparison with PRO. IMI induced a strong dose-response impact on the genes linked to the detoxification, ROS production, cell cycle, and apoptosis regulation. In conclusion, our results suggest that current pesticide concentrations detected in the Arcachon Bay are safe for the Pacific oyster early development, but they might have a small direct effect via altered gene expressions, whose longer-term impacts cannot be ruled out., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The distribution of the Mediterranean mussel Mytilus galloprovincialis extends more and more northwards in the Atlantic. Crossings are frequently observed with the blue mussel Mytilus edulis along the French and English coasts. The aim of this study is firstlyto identify the co-presence of M. galloprovincialis, M. edulis, and their hybrids in different sites of the Mediterranean and Atlantic coasts, and to provide insights for the thermal tolerance and toxicant susceptibility of Mytilus edulis, Mytilus galloprovincialis and their hybrids. Mussels were collected from the shore at 20 sampling sitesin Europe and Tunisia and identified using Me 15/16 primers targeting the adhesive protein gene sequence. Samples were screened for the presence of Mytilus edulis, Mytilus galloprovincialis, and hybrids alleles using PCR. To get more information on hybrids sensitivities to temperature and metals, freshly fertilized eggs of the two species and their hybrids were reared at four temperatures 18, 20, 22, and 24 °C and exposed to concentrations of Cu, Ag, and a mixture of both metals. Arrests of development and malformations were recorded after 48 h of exposure. The genotypic identification of the two species on 20 sites of the Mediterranean and Atlantic coasts carried out during this study confirms the presence of pure and hybrid species of mussel. Our results highlighted that hybrid larvae from a female of M. galloprovincialis are significantly more tolerant to temperature increases than pure larvae of M. galloprovincialis and pure and hybrid larvae of M. edulis. No significant interspecies-differences of sensitivity were noted for metal exposure alone. However, a co-exposure of larvae to both metal and high temperature highlighted the higher tolerance of hybrid larvae from a female of M. galloprovincialis to both stresses. The overall results could allow the prediction of the future evolution of mussel populations facing environmental changes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Microplastics are ubiquitous in aquatic ecosystems, but little information is currently available on the dangers and risks to living organisms. In order to assess the ecotoxicity of environmental microplastics (MPs), samples were collected from the beaches of two islands in the Guadeloupe archipelago, Petit-Bourg (PB) located on the main island of Guadeloupe and Marie-Galante (MG) on the second island of the archipelago. These samples have a similar polymer composition with mainly polyethylene (PE) and polypropylene (PP). However, these two samples are very dissimilar with regard to their contamination profile and their toxicity. MPs from MG contain more lead, cadmium and organochlorine compounds while those from PB have higher levels of copper, zinc and hydrocarbons. The leachates of these two samples of MPs induced sublethal effects on the growth of sea urchins and on the pulsation frequency of jellyfish ephyrae but not on the development of zebrafish embryos. The toxic effects are much more marked for samples from the PB site than those from the MG site. This work demonstrates that MPs can contain high levels of potentially bioavailable toxic substances that may represent a significant ecotoxicological risk, particularly for the early life stages of aquatic animals., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

The role of polyethylene microplastics 4-6 μm size (MPs) in the toxicity of environmental compounds to fish early life stages (ELS) was investigated. Marine medaka Oryzias melastigma embryos and larvae were exposed to suspended MPs spiked with three model contaminants: benzo(a)pyrene (MP-BaP), perfluorooctanesulfonic acid (MP-PFOS) and benzophenone-3 (MP-BP3) for 12 days. There was no evidence of MPs ingestion but MPs agglomerated on the surface of the chorion. Fish ELS exposed to virgin MPs did not show toxic effects. Exposure to MP-PFOS decreased embryonic survival and prevented hatching. Larvae exposed to MP-BaP or MP-BP3 exhibited reduced growth, increased developmental anomalies and abnormal behavior. Compared to equivalent waterborne concentrations, BaP and PFOS appeared to be more embryotoxic when spiked on MPs than when alone in seawater. These results suggest a relevant pollutant transfer by direct contact of MPs to fish ELS that should be included in the ecotoxicological risk assessment of MPs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The present work aims to investigate the effects of water temperature increase on Mytilus galloprovincilis and Mytilus edulis pure larvae (PG, PE) and their hybrids (HFG, HFE). D-larvae were maintained at 18 °C or exposed to a higher temperature of 22 °C for 48 h. Initially, Embryotoxicity test was evaluated. Second, a transcriptomic analysis using a recently developed microarray platform was applied to determine the main biological processes involved in early life stages responses to temperature increase. Finally, an immunofluorescence investigation was performed to bridge the gap between transcriptomic regulation and the real changes at cellular/tissue levels. Embryotoxicity test revealed a higher sensitivity of M. edulis (PE) D-larvae as well as hybrids from females M. edulis (HFE) to temperature increase, with the highest rate of larval malformations. Transcriptomic results indicated a lack of an adequate heat shock protein (Hsp) response in PE and HFE larvae (the high expression was observed in PG larvae); the differential expression of gene involved in translation, energy metabolism and oxidative stress response may contribute to explain the observed complex alterations in the studied conditions. As revealed by immunohistochemistry, cytoskeleton proteins changes associated with a drastic decrease of Histidine-Rich Glycoprotein (HRG) may elucidate the larval abnormalities in shell development observed for PE and HFE larvae. Overall, the results indicate that each type of pure larva (PG and PE) and their respective female hybrid (HFG and HFE) react similarly to the temperature increase. Our data should be carefully considered in view of the water temperature increase in marine ecosystems and especially for the mussel's species in confluence zones., (Copyright © 2019 Elsevier B.V. All rights reserved.)

This study investigates the combined effects of increased temperatures and copper on the early live stages of the Mediterranean mussel Mytilus galloprovincialis and the blue mussel Mytilus edulis as well as their hybrids. For this purpose, developmental abnormalities was measured after 48 h of exposure as well as a battery of oxidative stress markers such as, antioxidant enzyme activities, lipid peroxidation measured as malondialdehyde accumulation (MDA) and metallothionein contents (MT) and related gene expression pattern. Embryotoxicity and metal accumulation in tissues of mussel larvae exposed to a sublethal concentration of copper (10 μg/L) along with a slight temperature increase from 18 °C to22 °C were significantly increased after 48 h of exposure. Co-exposure to Cu and elevated temperatures significantly increased the antioxidant enzyme activities termed as, glutathione-S-transferase (GST), catalase (CAT), superoxide dismutase (SOD) and being maintained at 22 °C only in pure M. galloprovincialis (PG) larvae and female galloprovincialis x male edulis hybrid larvae (HFG). A significant decrease in mRNA abundance of cat, sod, gst gene transcription levels was showed in the same species. Furthermore, metallothionein accumulation increased significantly in PG D-larvae exposed to copper at 22 °C. The same pattern was observed in term of gene expression of MTs cognates (mt-10 and mt-20). Significant increase of MDA levels in pure M. edulis (PE) larvae and their female M. edulis x male M. galloprovincialis hybrid larvae (HFE) combined with a low MTs content were observed. Overall, this study provides clues about the relatively higher resistance and resilience of M. galloprovincialis species compared to M. edulis species under environmental pollution and future climate change scenarios., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The present study aims to evaluate the effects of copper and silver alone or along with a moderate temperature increase on embryonic development, DNA integrity and target gene expression levels in early life stages of Mytilus galloprovincialis. For this purpose, upon fertilized embryos were exposed to a sub-lethal concentration of Cu (9.54 μg/L), Ag (2.55 μg/L) and to the mixture of the two metals (Cu (6.67 μg/L) + Ag (1.47 μg/L)) along with a temperature gradient (18, 20 and 22 °C). In all experiments, larvae were exposed to stressors for 48 h except for those designed to DNA damage analysis exposed only for 24 h (before shell formation).Our results showed a significant increase in the percentage of malformed D-larvae (p < 0.05) with increasing temperature and exposure to silver and copper alone or in a mixture. Moreover, metal toxicity increased significantly (p < 0.05) with the temperature rise. Genotoxicity was evaluated using classic and modified with Formamidopyrimidine DNA glycosylase (Fpg) Comet assay. Results suggest that co-exposure to metals and temperature significantly increased DNA damage on mussel larvae with a more accentuated oxidative damage. A significant transcription modulation was observed for genes involved in DNA repair and DNA replication (p53, DNA ligase II and topoisomerase II) when larvae are exposed to a single stressor. However, in the case of multiple stresses, caspase involved in the cell apoptosis pathway was overexpressed. Our study suggests that mussel larvae exposed to a moderate increase in temperature may have a compromised ability to defend against genotoxicity. This is particularly relevant in the context of global warming and thermal pollution., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The aim of this work was to study the impact of copper during a sub-chronic exposure to environmental concentrations in the early life stages of rainbow trout (Oncorhynchus mykiss). Eyed-stage embryos of rainbow trout, at 265 °D, were exposed in semi-static conditions to sub-lethal concentrations of CuSO 4 up to the larval stage (528 °D) under laboratory-controlled conditions. During 3 weeks, they were exposed to the environmentally-realistic concentration of 2 µg/L Cu and to a 10-fold higher concentration, 20 µg/L Cu. Several biological (survival, hatching success, malformation, growth) and behavioral (swimming activity) and molecular endpoints (genotoxicity and gene transcription) were studied. Exposure to 20 µg/L Cu had an inhibitory effect on hatching and increased half-hatched embryos (25%). At the end of the exposure, no significant differences were observed in growth of the larvae exposed to the highest Cu concentration. However, larvae exposed to 2 µg/L Cu exhibited increased growth in comparison with non-exposed larvae. The percentage of malformed larvae was significantly higher for both copper conditions, with skeletal malformations being the most observed. Expression of several genes was evaluated in whole larvae using quantitative real-time PCR. Genes involved in detoxification (gst, mt1 and mt2) and in cell cycle arrest (p53) were significantly repressed in both copper conditions when compared to control. In addition, potential genotoxic effects on larvae were investigated by the comet assay on blood cells, but this test did not demonstrate any significant DNA damage on larvae exposed to copper. This study confirms the adverse effects of copper on early life stages of rainbow trout even at the lowest environmentally relevant tested concentration., (Copyright © 2018 Elsevier Inc. All rights reserved.)

It is now well documented that several contaminants can modulate the fish immune system, leading to disrupted host resistance against pathogens and increased incidence of disease. Since fish are usually co-exposed to chemicals and pathogens in the natural environment, analysis of the immunotoxic effects of pollutants is particularly relevant. The authorities in the European Union have recommended the development of toxicity assays on cell cultures and embryos, as an alternative to testing in vertebrates. This is why in our study, a fish immune challenge assay was developed for the early life stages of Japanese medaka to evaluate and compare the relevance of new biomarkers. Fish were exposed to benzo[a]pyrene (BaP), a model pollutant, for 8days at the embryonic stage, or for 48h at the larvae and juvenile stages, and fish were infected with betanodavirus by bath-challenge of 10 6 TCID 50 /mL. Biometric changes and induction of malformations were observed after embryonic exposure. DNA damage and induction of EROD activity were recorded at the end of all chemical exposures. Viral infection increased the mortality rate significantly and disturbed the behavior of fish after light stimulation. While BaP exposure increased swimming speed, betanodavirus infection slowed swimming activity. In larvae co-exposed to BaP and the virus, the viral titer in the whole body was higher than in fish infected only with the virus. This study highlighted the sensitivity and usefulness of the immune challenge assay on the early life stages of Japanese medaka to evaluate the toxic effects of pollutants., (Copyright © 2018. Published by Elsevier B.V.)

Toxicity of polyethylene microplastics (PE-MP) of size ranges similar to their natural food to zooplanktonic organisms representative of the main taxa present in marine plankton, including rotifers, copepods, bivalves, echinoderms and fish, was evaluated. Early life stages (ELS) were prioritized as testing models in order to maximize sensitivity. Treatments included particles spiked with benzophenone-3 (BP-3), a hydrophobic organic chemical used in cosmetics with direct input in coastal areas. Despite documented ingestion of both virgin and BP-3 spiked microplastics no acute toxicity was found at loads orders of magnitude above environmentally relevant concentrations on any of the invertebrate models. In fish tests some effects, including premature or reduced hatching, were observed after 12 d exposure at 10 mg L -1 of BP-3 spiked PE-MP. The results obtained do not support environmentally relevant risk of microplastics on marine zooplankton. Similar approaches testing more hydrophobic chemicals with higher acute toxicity are needed before these conclusions could be extended to other organic pollutants common in marine ecosystems. Therefore, the replacement of these polymers in consumer products must be carefully considered., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Fish are currently used for the assessment of chemical toxicity. The REACh regulation and the European directive on the protection of animals used for scientific purposes both recommend the use of methods other than animal testing. In view of this, fish cell lines are increasingly used to provide fast and reliable toxic and ecotoxic data on new chemicals. The sensitivity of the Rainbow trout liver cell line RTL-W1 and Japanese medaka embryos cell line OLCAB-e3 were used with different toxicity endpoints, namely cytotoxicity, EROD activity, ROS production and DNA damage for various classes of pollutants displaying different modes of action but also with complex environmental mixtures. Toxicity tests were coupled with chemical analysis to quantify the chemical concentrations in cell cultures. Differences in sensitivity were found between fish cell lines. MTT reduction assay revealed that OLCAB-e3 cells were more sensitive than RTL-W1 cells. On the contrary, RTL-W1 gave higher response levels for the Fpg-modified comet assay and ROS assay. The OLCAB-e3 cell line did not express EROD activity unlike RTL-W1. This study highlights the capacity of the two different fish cell lines to measure the toxicity of individual toxicants but also environmental mixtures. Then, results obtained here illustrate the interest of using different cell lines and toxicity endpoints to assess the toxicity of complex or unknown mixture of chemicals., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Sediment compartment is a long term sink for pollutants and a secondary source of contamination for aquatic species. The abiotic factors controlling the bioavailability and thus the toxicity of complex mixtures of pollutants accumulated in sediments are poorly documented. To highlight the different factors influencing sediment toxicity, we identified and analyzed the physico-chemical properties, micro-pollutant contents, and toxicity level of six contrasted sediments in the Lot-Garonne continuum. Sediment toxicity was evaluated using the recently described Japanese medaka (Oryzias latipes) embryo-larval assay with direct exposure to whole sediment (MELAc). Multiple toxicity endpoints including embryotoxicity, developmental defects and DNA damage were analyzed in exposed embryos. Chemical analyses revealed significant variations in the nature and contamination profile of sediments, mainly impacted by metallic trace elements and, unexpectedly, polycyclic aromatic hydrocarbons. Exposure to sediments induced different toxic impacts on medaka early life stages when compared with the reference site. Principal component analysis showed that the toxic responses following exposure to sediments from the Lot River and its tributary were associated with micro-pollutant contamination: biometric measurements, hatching success, genotoxicity, craniofacial deformities and yolk sac malabsorption were specifically correlated to metallic and organic contaminants. Conversely, the main biological responses following exposure to the Garonne River sediments were more likely related to their physico-chemical properties than to their contamination level. Time to hatch, cardiovascular injuries and spinal deformities were correlated to organic matter content, fine particles and dissolved oxygen levels. These results emphasize the necessity of combining physico-chemical analysis of sediment with toxicity assessment to accurately evaluate the environmental risks associated with sediment contamination., (Copyright © 2017 Elsevier Inc. All rights reserved.)

The present study aims to elucidate the stress response of early life stages of Mytilus galloprovincialis to the combine effects of selected metals and elevated temperature. For this purpose, we investigated the response of a large panel of oxidative stress markers such as catalase (CAT), superoxide dismutase (SOD), glutathione-S-transferase (GST) activities and lipid peroxidation (thiobarbituric acid reactive substrates (TBARS) concentration) and metallothionein accumulation (MT) as well as selected gene transcription level and metal accumulation in mussels larvae exposed to a sub-lethal concentration of Cu (9.54µg/L), Ag (2.55µg/L) and mixture of the two metals (Cu (6.67µg/L)+Ag (1.47µg/L)) along with a temperature gradient (18, 20 and 22°C) for 48h. Cu and Ag applied as single or mixture were differentially accumulated in mussel larvae according to the exposure temperature. Sod, cat, gst and mt-10 gene transcription levels showed an important increase in larvae exposed to Cu, Ag or to the mix compared to the control condition at 18°C. The same pattern but with higher induction levels was recorded in larvae co-exposed to metals at 20°C. At 22°C, a significant decrease in mRNA abundance of cat, gst and sod and a significant up-regulation of mts targets (mt10 and mt20) were observed., Results: suggest that co-exposure to metals and moderate elevated temperature (20 and 22°C) significantly increased the antioxidant enzyme activities of catalase (CAT), and glutathione-S-transferase (GST) and caused an increase of metal and metallothionein concentrations. In contrast, no significant change in lipid peroxidation products measured as TBARS content was observed indicating a protective response of anti-oxidative system. This study provides first evidences of the early and efficient protective response of antioxidant defense mechanisms in mussel's early life stages facing in multi stressors situations., (Copyright © 2016. Published by Elsevier Inc.)

The International workshop on Integrated Assessment of CONtaminants impacts on the North sea (ICON) provided a framework to validate the application of chemical and biological assessment thresholds (BACs and EACs) in the Seine Bay in France. Bioassays (oyster larval anomalies, Corophium arenarium toxicity assay and DR Calux) for sediment and biomarkers: ethoxyresorufin-O-deethylase (EROD) activity, acetylcholinesterase (AChE) activity, lysosomal membrane stability (LMS), DNA strand breaks using the Comet assay, DNA adducts, micronucleus (MN), PAH metabolites, imposex, intersex and fish external pathologies were analysed in four marine sentinel species (Platichthys flesus, Limanda limanda, Mytilus sp. and Nucella lapilus). Polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and heavy metals were analysed in biota and sediment. Results for sediment and four species in 2008-2009 made it possible to quantify the impact of contaminants using thresholds (Environmental Assessment Criteria/EAC 2008 : 70% and EAC 2009 : 60%) and effects (EAC 2008 : 50% and EAC 2009 : 40%) in the Seine estuary. The Seine estuary is ranked among Europe's most highly polluted sites., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

This study evaluates the combined effects of two contaminants (copper and a herbicide S-metolachlor) and temperature on the early life stages of the Pacific oyster Crasssotrea gigas, which is native to Arcachon Bay (southwest France). The responses of D-larvae, obtained from wild and cultivated oysters, were investigated for one year during the oyster breeding period at different sampling sites and compared with the response of D-larvae from a commercial hatchery. Embryotoxicity was measured by considering the percentage of abnormal D-shaped larvae. Normal development of embryos occurred only from 22°C to 26°C. There were synergistic effects of copper at low and high temperatures. Native oysters appear particularly sensitive to an increase in water temperature, suggesting a future increase in the percentage of larval abnormalities as a result of global climate change. Hatchery oysters represent a good alternative model for studying the effects of both pollutants and climate change stressors., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The present work aimed to assess the effects of two widespread metallic pollutants, copper and silver, along with environmentally-realistic temperature increases, on embryo-larval development of the Mediterranean mussel Mytilus galloprovincialis. First, mussel embryos upon fertilization were exposed for 48 h to increasing concentrations of Cu (0.5-500 μg/L) and Ag (0.1-100 μg/L) at different temperatures (18, 20, 22 or 24 °C) in order to characterize toxicity of each toxicant at the different tested temperatures. Increasing concentrations of a Cu-Ag mixture were then tested in order to assess the mixture effect at different temperatures (18, 20 or 22 °C). Embryotoxicity was measured after 48 h of exposure (D-larvae stage) considering both the percentage of abnormalities and developmental arrest in D-larvae. The results suggest that the optimum temperature for mussel larvae development is 18 °C (12.65± 1.6% malformations) and beyond 20 °C a steep increase of abnormal larvae was observed up to 100% at 24 °C. Ag was more toxic than Cu with a 50% effective concentration (EC50) at 18 °C of 6.58 μg/L and 17.6 μg/L, respectively. Temperature increased the toxicity of both metals as proved with the EC50 at 20 °C at 3.86 μg/L and 16.28 μg/L for Ag and Cu respectively. Toxic unit calculation suggests additive effects of Cu and Ag in mixture at 18 and 20 °C. These results highlight a possible impairment of M. galloprovincialis reproduction in the Mediterranean Sea in relation to increase of both pollutants and water temperature due to global warming., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Petroleum compounds from chronic discharges and oil spills represent an important source of environmental pollution. To better understand the deleterious effects of these compounds, the toxicity of water-accommodated fractions (WAF) from two different oils (brut Arabian Light and Erika heavy fuel oils) were used in this study. Zebrafish embryos (Danio rerio) were exposed during 96h at three WAF concentrations (1, 10 and 100% for Arabian Light and 10, 50 and 100% for Erika) in order to cover a wide range of polycyclic aromatic hydrocarbon (PAH) concentrations, representative of the levels found after environmental oil spills. Several endpoints were recorded at different levels of biological organization, including lethal endpoints, morphological abnormalities, photomotor behavioral responses, cardiac activity, DNA damage and exposure level measurements (EROD activity, cyp1a and PAH metabolites). Neither morphological nor behavioral or physiological alterations were observed after exposure to Arabian Light fractions. In contrast, the Erika fractions led a high degree of toxicity in early life stages of zebrafish. Despite of defense mechanisms induced by oil, acute toxic effects have been recorded including mortality, delayed hatching, high rates of developmental abnormalities, disrupted locomotor activity and cardiac failures at the highest PAH concentrations (∑TPAHs=257,029±47,231ng·L(-1)). Such differences in toxicity are likely related to the oil composition. The use of developing zebrafish is a good tool to identify wide range of detrimental effects and elucidate their underlying foundations. Our work highlights once more, the cardiotoxic action (and potentially neurotoxic) of petroleum-related PAHs., (Copyright © 2016 Elsevier B.V. All rights reserved.)

For several years, low larval recruitment has been observed in Arcachon Bay, in southwest France. Exposure to pollutants could partly account for the reduction of early life stages of the Pacific oyster. This study evaluated the effects of copper and S-metolachlor in combination with salinity on the early life stages of Crassostrea gigas. Embryos were exposed to concentrations of copper (1, 10 and 50 μg L(-1)) or S-metolachlor (10, 100 and 1000 ng L(-1)) and six salinities (18, 21, 24, 27, 30 and 33 u.s.i). Embryotoxicity was measured by considering both the percentage of abnormalities and arrested development in D-shaped larvae. Embryo-larval development was only affected at salinities ≤24 u.s.i, which have never been observed during C. gigas reproduction period in Arcachon Bay. Both contaminants had an effect at environmental concentrations. Our results suggest that copper and metolachlor toxicity was enhanced with decreasing salinity., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Benz[a]anthracene (BaA) is a ubiquitous polycyclic aromatic hydrocarbon found in numerous aquatic ecosystems. However, ecotoxicological data in aquatic organisms are scarce. To remedy this lack of data, Japanese medaka (Oryzias latipes) embryos were exposed to BaA and toxic effects were investigated at multiple toxicological endpoints. Japanese medaka embryos were incubated onto BaA-spiked artificial sediment for 9 days at low or moderate environmental concentrations ranging from 0.9 to 12 µgg(-1) dw. BaA-exposed embryos exhibited significant tachycardia. BaA exposure was also shown to increase CYP1A activity in the hepato-biliary tissue as well as craniofacial deformities and DNA damage in pro-larvae. The photomotor response of BaA-exposed larvae was reduced in comparison to the control group. According to this set of tests, the lowest tested and observed effect concentration (LOEC) for Japanese medaka early life stages was equivalent to 0.92 µgg(-1) dw of BaA. This concentration fall into the range of concentrations frequently encountered in sediments of polluted aquatic ecosystems. Taking into consideration these results, BaA represents a threat for fish early life stages in particular those developing onto or into contaminated sediments., (Copyright © 2014 Elsevier Inc. All rights reserved.)

In this study, biological responses and contaminant levels in biological tissues were investigated in grass goby fish specimens (Zosterisessor ophiocephalus) collected from five stations in a moderately polluted ecosystem, namely the Bizerte lagoon on the north coast of Tunisia. The following biomarkers were measured: muscular acetylcholinesterase (AChE), hepatic ethoxyresorufin-O-deethylase (EROD), glutathione-S-transferase (GST), catalase (CAT), lipoperoxidation (TBARS), condition factor (CF), and hepatosomatic index (HSI). These measurements were taken in parallel with the content of Organochlorine pesticides (OCPs), Polychlorinated biphenyls (PCBs), Polycyclic aromatic hydrocarbons (PAHs) and trace metals (As, Cr, Cu, Mn, Pb, V, Zn, Ag, Cd, Co and Ni) in muscle tissue. Total PAH concentrations ranged from 20.09 ± 0.68 to 105.77 ± 42.58 ng g(-1) dw, PCB from 33.19 ± 6.25 to 126.28 ± 7.37 ng g(-1) dw, OCP from 11.26 ± 1.62 to 19.17 ± 2.06 ng g(-1) dw, and metals from 107.83 ± 1.83 to 187.21 ± 2.00 mg/kg dw. The highest levels of pollutants and biomarkers were observed at station S1, located in the Bizerte channel. Elevated EROD, GST and CAT activities, as well as TBARS levels in liver were positively correlated with tissue contaminant levels at station S1. Significant negative correlations were also found between hexachlorobenzene (HCB) and dichlorodiphenyltrichloroethane (DDTs) body burden with AChE activity in muscle at station S2. The integration of biological responses and contaminant tissue content indicated that certain areas of the Bizerte lagoon, notably station S1, are significantly impacted by various human activities, which likely represent a threat for aquatic wildlife. On the basis of these results, and due to its ecological characteristics, the grass goby appears a suitable indicator species for pollution biomonitoring in coastal marine areas along the Mediterranean Sea., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

In order to assess the environmental quality of the Bizerte lagoon (Tunisia), biomarker and contaminant levels were measured in Mediterranean mussels (Mytilus galloprovincialis) from five selected sites. Persistent organic pollutants (POPs) were quantified in whole body and enzyme activities such as acetylcholinesterase (AChE), catalase (CAT) and glutathione S-transferase (GST) in gills. Despite the relatively low levels of organic contaminants, the selected biomarkers responded differently according to the pollution level at the different sites. GST and AChE activities were correlated with the amount of DDTs in mussel tissues. These two enzymatic activities were also correlated to temperature and pH. No significant difference was observed for CAT activity. Principal component analysis showed a clear separation of sampling sites in three different assemblages which is consistent with POP body burden in mussels. Our results confirmed the usefulness of combining biomarker and chemical analyses in mussels to assess chemical pollution in the Bizerte lagoon., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Irgarol and diuron are the most representative "organic booster biocides" that replace organotin compounds in antifouling paints, and metolachlor is one of the most extensively used chloroacetamide herbicides in agriculture. The toxicity of S-metolachlor, irgarol and diuron was evaluated in Pacific oyster (Crassostrea gigas) gametes or embryos exposed to concentrations of pesticides ranging from 0.1× to 1000×, with 1× corresponding to environmental concentrations of the three studied pesticides in Arcachon Bay (France). Exposures were performed on (1) spermatozoa alone (2) oocytes alone and (3) both spermatozoa and oocytes, and adverse effects on fertilization success and offspring development were recorded. The results showed that the fertilizing capacity of spermatozoa was significantly affected after gamete exposure to pesticide concentrations as low as 1× of irgarol and diuron and 10× of metolachlor. The offspring obtained from pesticide-exposed spermatozoa displayed a dose-dependent increase in developmental abnormalities. In contrast, treating oocytes with pesticide concentrations up to 10× did not alter fertilization rate and offspring quality. However, a significant decline in fertilization success and increase in abnormal D-larvae prevalence were observed at higher concentrations 10× (0.1 μg L(-1)) for S-metolachlor and 100× for irgarol (1.0 μg L(-1)) and diuron (4.0 μg L(-1)). Irgarol, diuron and S-metolachlor also induced a dose-dependent increase in abnormal D-larvae prevalence when freshly fertilized embryos were treated with pesticide concentrations as low as concentration of 1× (0.01 μg L(-1) for irgarol or S-metolachlor, and 0.04 μg L(-1) for diuron). The two bioassays on C. gigas spermatozoa and embryos displayed similar sensitivities to the studied pesticides while oocytes were less sensitive. Diuron, irgarol and S-metolachlor induced spermiotoxicity and embryotoxicity at environmentally relevant concentrations and therefore might be a threat to oyster recruitment in coastal areas facing chronic inputs of pesticides., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Hydrophobic pollutants, in particular sediment-sorbed organic compounds, are widespread in the aquatic environment and could represent a threat to living organisms. Estuarine species, which live in turbulent ecosystems, are particularly exposed to this mode of contamination. For precise evaluation of the toxicity of hydrophobic contaminants desorbed from particles, a new larval assay using nauplii of the estuarine calanoid copepod Eurytemora affinis was developed. It consists of the direct exposure of copepods during naupliar development to elutriates of an unpolluted sediment spiked with different model contaminants. This bioassay measures the toxicity of the bioavailable fraction of particle-sorbed pollutants on the naupliar stage of copepods. Mortality and growth (non-invasive endpoints) in nauplii were analysed after six days of exposure. This approach was validated using six pollutants with different modes of action: benzo[a]pyrene (BaP), dimethylbenzo[a]anthracene (DMBA), phenanthrene (PHE), polychlorinated biphenyls (PCB 126, PCB 153) and 4-nonylphenol (4-NP). All these compounds induced a dose-dependent increase in toxic effects. Lethal effects only occurred at the highest tested concentrations: 58,541 and 6092 ng g(-1) dry weight sediment (dws), for PHE and DMBA, respectively. Sublethal effects (growth inhibition) were observed at lower concentrations for all tested compounds except PCB 153, from 8, 142, 297, 6092 and 8453 ng g(-1) dws for PCB 126, BaP, PHE, DMBA and 4-NP, respectively., (Copyright © 2013 Elsevier Inc. All rights reserved.)

This study evaluated embryotoxicity and genotoxicity of two dissolved metals copper and cadmium (Cu and Cd) and two pesticides (metolachlor and irgarol) occurring in Arcachon Bay (SW France) in Pacific oyster (Crassostrea gigas) larvae and investigated the relationship between those two endpoints. Embryotoxicity was measured by calculating the percentage of abnormal D-shaped larvae and genotoxicity was evaluated with DNA strand breaks using the comet assay. After 24h exposure, significant increases of the percentage of abnormal D-larvae and the DNA strand breaks were observed from 0.1 μg L⁻¹ for Cu, 10 μg L⁻¹ for Cd and 0.01 μg L⁻¹ for both irgarol and metolachlor in comparison with the controls. A strong positive relationship between embryotoxicity and genotoxicity was recorded for Cu, Cd and metolachlor. The current study suggests that copper, irgarol and metolachlor can induce larval abnormalities and DNA damage in a population of exposed oysters at environmentally relevant concentrations., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Because of their high capacity to accumulate contaminants such as persistent organic pollutants and heavy metals, aquatic sediments are considered as a long-term source of contamination for aquatic organisms. In compliance with the increasing interest both for sediment quality evaluation and the use of fish early life stage (ELS) toxicity assays, we proposed an embryo-larval test to evaluate embryotoxicity and genotoxicity of sediment-bound contaminants. Pre-blastula stage medaka (Oryzias latipes) embryos were exposed by static sediment contact to two model heavy metals (cadmium and copper) at environmental concentrations during the whole 10-day embryonic development. Lethal and sub-lethal effects were recorded in both embryos and larvae for 20 days post fertilisation (dpf) using several global toxicity and phenotypic endpoints. The comet assay was also performed on medaka prolarvae to evaluate genotoxic effects of the tested chemicals. Environmental concentrations of cadmium (Cd) and copper (Cu) did not affect embryo and larval survival. However, both heavy metals significantly induced morphological abnormalities, particularly spinal and cardiovascular deformities. Cd but not Cu induced tachycardia. Both heavy metals induced a significant increase in DNA damage at all tested concentrations. Resulting LOEC values for Cd and Cu corresponded to 1.9 and 8.5 μg/g d.w. sediment, respectively. Although metal bioavailability is probably lower for naturally contaminated sediments, the relatively low toxicity thresholds for both Cd and Cu raise the question of possible risk for fish embryos developing in direct contact to sediments. This study demonstrates the applicability, sensitivity and relevance of the Japanese medaka embryo-larval assay (MELA) to evaluate sediment hazardous potency at environmental concentrations of heavy metals., (Copyright © 2012. Published by Elsevier Inc.)

In this study, new DNA markers were explored for the flounder Platichthys flesus. cDNA and genomic sequences of the genes encoding the glyceraldehyde-3-phosphate-deshydrogenase (GAPDH), the cytosolic creatine kinase (CK), the prostaglandin D synthase (PGDS) and the betaine homocysteine methyltransferase (BHMT) were characterized. The tumour suppressor p53 gene structure was already described. A PCR-SSCP (Single Strand Conformation Polymorphism) analysis was finally conducted to study the genetic polymorphism of different populations of flounders collected along the French Atlantic coast. Four highly contaminated French estuaries (Seine, Vilaine, Loire and Gironde) were sampled and compared to a reference estuary (Ster) to explore possible selective effect of the environment on specific allelic frequencies. Our results showed that two loci p53 and PGDS, could be potential markers of chemical stress: p53A allele frequency increased in contaminated systems compared to the reference system. In the Vilaine estuary, PGDS polymorphism could be related to pesticide stress.

A genetically engineered diploid yeast strain named yJC2, was specifically developed for environmental mutagen detection and characterization of induced mutations. This strain contains one copy of the human TP53 tumour suppressor gene coding sequence which is used as a molecular target for mutagens and two copies of the ADE2 reporter gene allowing accurate measurement of the TP53 transcriptional activity. The strain sensitivity to mutagens was evaluated by exposing cells to UVC, 4-nitroquinoline (NQO) or to an organic extract of sediment from the Seine estuary. For all studied mutagens, a significant and dose-dependent increase of mutant frequency was observed. The present assay named FACIM II (Functional Analysis of Chemical-Induced TP53 Mutations) is more convenient than the FACIM I and more inducible than the SOS Chromotest to detect direct-acting mutagens in the environment.

A functional assay was developed in yeast to identify mutations induced by DNA-damaging agents at the flounder TP53 locus. This assay named FACIM for functional analysis of chemically-induced p53 mutations, is based on the assumption that most genotoxin-induced mutations inactivate transcriptional activity of the TP53 protein. The functional status of the protein expressed in yeast was measured using a p53-responsive reporter gene. The FACIM assay was used to evaluate the mutagenesis of the flounder TP53 exposed in vitro to benzo[a]pyrene diol epoxide (BPDE). A dose-dependent increase of p53 mutation rate was observed with increasing concentrations of BPDE and extension of exposure time. Flounder TP53 gene appeared highly sensitive to point mutations since most of those identified targeted different nucleotides. Mutated base-pairs corresponded predominantly to guanines located on the non-transcribed strand of the DNA. The general distribution of mutations along the flounder TP53 protein was different from that identified in the human homologue suggesting species-differences in mutagenesis of the TP53 gene. Most of flounder TP53 mutants were defective for transactivation and cell growth regulation but some maintained a partial wild-type phenotype. This functional assay in yeast could be used for both evaluation of the genotoxic potency of chemicals or environmental samples and screening of p53 mutations in fish tumours.

A number of flounders dwelling in highly contaminated coastal areas of Northern Europe develop liver tumours. In order to increase our understanding of the molecular pathogenesis of these sporadic tumours, we examined p53 mutations in eleven hyperplasia and six adenoma. p53 introns 4 to 8 were first sequenced to allow individual amplification of exons 5 to 8. DNA extracted from formalin-fixed livers was amplified and PCR products were directly sequenced. Two major results were obtained. (i) Flounders from different geographical areas displayed a high rate of sequence variation. Base substitutions were identified in both tumour and normal tissues and thus may be considered as polymorphic variations in individuals. (ii) One mutation was detected in two hyperplastic foci from the same flounder. This mutation was a T:A to A:T transversion at codon 147, resulting in the replacement of valine for glutamic acid. This residue took place in the L2 loop of the DNA binding surface. Its substitution by an hydrophilic and charged residue could thus impair p53 (protein) biological activity.