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Many indicators have been developed to assess the state of benthic communities and identify seabed habitats most at risk from bottom trawling disturbance. However, the large variety of indicators and their development and application under specific geographic areas and management contexts has made it difficult to evaluate their wider utility. We compared the complementarity/uniqueness, sensitivity, and selectivity of 18 benthic indicators to pressure of bottom trawling. Seventeen common datasets with broad regional representation covering a range of pressure gradients from bottom trawling disturbance (n = 14), eutrophication (n = 1), marine pollution (n = 1), and oxygen depletion (n = 1) were used for the comparison. The outcomes of most indicators were correlated to a certain extent with response to bottom trawling disturbance, and two complementary groups of indicators were identified: diversity-based and biological trait-based indicators. Trait-based indicators that quantify the changes in relative abundance of sensitive taxa were most effective in identifying benthic community change in response to bottom trawling disturbance. None of the indicators responded to the trawling pressure gradient in all datasets, and some showed a response that were opposed to the theoretical expectation for some gradients. Indicators that showed clear responses to bottom trawling disturbance also showed clear responses in at least one other pressure gradient, suggesting those indicators are not pressure specific. These results emphasize the importance of selecting several indicators, at least one from each group (diversity and trait-based), to capture the broader signals of change in benthic communities due to bottom trawling activities. Our systematic approach offers the basis from which scientific advisors and/or managers can select suitable combinations of indicators to arrive at a sensitive and comprehensive benthic status assessment., (© 2024 The Author(s). Ecological Applications published by Wiley Periodicals LLC on behalf of The Ecological Society of America.)

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.)

Lately, the role of microplastics (MP) as vectors for dissolved contaminants and as vehicle for their transfer to aquatic organisms has received attention. Similarly to MP, other inorganic and organic particles may act as passive samplers. However, limited comparative knowledge exists at this respect. In the present study we have comparatively investigated the risk for mussel of MP and the pesticide chlorpyrifos (CPF) alone and in combination with MP and phytoplankton particles of microalgae (MP-CPF and MA-CPF, respectively). We selected MP and microalgae of similar size to expose mussel to the same volume of particles (≈1.5 mm 3 L -1 ≈ equivalent to 1.5 mg MP L -1 ) and the same concentration of contaminant (CPF, 7.6 μg L -1 ). MP were virgin HDPE microparticles (≤10 μm) while the microalgae species was Isochrisis galbana (4-8 μm). Mussels were exposed for 21 days to MP, CPF, MP-CPF and MA-CPF. Then, a suite of neurotoxicity, oxidative stress and oxidative damage biomarkers were measured in samples collected at day 7 and 21. Additionally, these biochemical markers were assessed in an integrated manner with others measured at physiological, immune and cell component level in the same organisms, previously published. Overall, MP did not elicit significant alterations on the majority of parameters measured. In contrast, mussels exposed to CPF, MA-CPF and MP-CPF showed evidence of neurotoxicity and oxidant imbalance at day 7, added to a detrimental physiological condition and immune imbalance at day 21. At the latter time MP-CPF mussels showed greater alterations than CPF or MA-CPF mussels. This suggested a synergistic toxicity of MP combined with CPF greater than that produced by the contaminants alone (MP or CPF) or by MA combined with CPF., 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. Published by Elsevier B.V.)

Microplastics (MPs) raise concerns not only as pollutants themselves, but also due to their ability to act as vectors of pollutants adsorbed from seawater, transferring them to marine organisms. However, the relevance of MPs as carriers of pollutants compared to microalgae needs further exploration. This study compared the role of MPs (2-10 μm non-oxidized and 10-15 μm oxidized high-density polyethylene) and natural organic particles (Rhodomonas lens microalgae, MA) as carriers of mercury (Hg, 2.3 μg Hg/L) and chlorpyrifos (CPF, 1.0 μg CPF/L) to adult Acartia tonsa copepods, after 24-48 h exposure. Dose-response experiments were first performed with adult female copepods exposed to oxidized MPs (0.25-4.0 mg/L), waterborne Hg (0.01-10.0 μg/L) and Ox MPs + Hg (0.25-4.0 mg oxidized MPs/L + 0.50-8.0 μg Hg/L) for 48 h, to complement previous studies that focused on the pesticide CPF. Effects were evaluated with four replicates for physiological and reproductive responses (6 females/replicate), biochemical techniques (40 individuals/replicate) and Hg/CPF bioaccumulation measurements (1000 individuals/replicate). Copepods accumulated Hg/CPF similarly from dissolved pollutants (6204 ± 2265 ng Hg/g and 1251 ± 646 ng CPF/g) and loaded MPs (3125 ± 1389 ng Hg/g and 1156 ± 266 ng CPF/g), but significantly less from loaded MA (21 ± 8 ng Hg/g and 173 ± 80 ng CPF/g). After 24-48 h, copepods exposed to MPs + Hg/CPF showed generally greater biological effects than those exposed to dissolved Hg/CPF or to MA + Hg/CPF, although differences were not statistically significant. MA + CPF had significantly lower AChE inhibition (1073.4 nmol min -1  mg -1 ) and MA + Hg lower GRx induction (48.8 nmol min -1  mg -1 ) compared to MPs + Hg/CPF and dissolved Hg/CPF (182.8-236.4 nmol min -1  mg -1 of AChE and 74.1-101.7 nmol min -1  mg -1 of GRx). Principal component analysis suggested different modes of action for Hg and CPF., 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.)

This study investigated the role of ultraviolet (UV) radiation and oxidation in high-density polyethylene microplastics (2-15 μm) and nanoplastics (0.2-9.9 μm) (NMPs) on particle chemistry, morphology, and reactivity with cadmium (Cd). Additionally, toxicity of NMPs alone and with Cd was evaluated using RTgutGC cells, a model of the rainbow trout (Oncorhynchus mykiss) intestine. The role on NMPs on Cd bioaccumulation in RTgutGC cells was also evaluated. Dynamic light scattering indicated that after UV radiation NPs agglomerated size increased from 0.8 to 28 µm, and to 8 µm when Cd was added. Oxidized MPs agglomerated size increased from 11 and 7 to 46 and 27 µm in non-UV- and UV-aged oxidized MPs when adding Cd, respectively. Cd-coated particles exhibited generally significantly higher zeta potential than non-Cd-coated particles, while attenuated total reflectance-Fourier transform infrared spectroscopy showed that the functional chemistry of the particles was oxidized and modified after being exposed to UV radiation. Presence of NMPs resulted in a significant decrease in Cd bioaccumulation in RTgutGC cells (100.5-87.9 ng Cd/mg protein) compared to Cd alone (138.1 ng Cd/mg protein), although this was not quite significant for co-exposures with UV-aged NPs (105.7 ng Cd/mg protein). No toxicity was observed in RTgutGC cells exposed to NMPs alone for 24 h. Moreover, co-exposures with Cd indicated that NMPs reduce the toxicity of Cd. Altogether these results show that UV aging enhances NMP surface reactivity, increasing Cd absorption in solution, which resulted in a reduction in Cd bioavailability and toxicity., (© 2024. The Author(s).)

This study investigates the combined impact of microplastics (MP) and Chlorpyriphos (CPF) on sea urchin larvae (Paracentrotus lividus) under the backdrop of ocean warming and acidification. While the individual toxic effects of these pollutants have been previously reported, their combined effects remain poorly understood. Two experiments were conducted using different concentrations of CPF (EC10 and EC50) based on previous studies from our group. MP were adsorbed in CPF to simulate realistic environmental conditions. Additionally, water acidification and warming protocols were implemented to mimic future ocean conditions. Sea urchin embryo toxicity tests were conducted to assess larval development under various treatment combinations of CPF, MP, ocean acidification (OA), and temperature (OW). Morphometric measurements and biochemical analyses were performed to evaluate the effects comprehensively. Results indicate that combined stressors lead to significant morphological alterations, such as increased larval width and reduced stomach volume. Furthermore, biochemical biomarkers like acetylcholinesterase (AChE), glutathione S-transferase (GST), and glutathione reductase (GRx) activities were affected, indicating oxidative stress and impaired detoxification capacity. Interestingly, while temperature increase was expected to enhance larval growth, it instead induced thermal stress, resulting in lower growth rates. This underscores the importance of considering multiple stressors in ecological assessments. Biochemical biomarkers provided early indications of stress responses, complementing traditional growth measurements. The study highlights the necessity of holistic approaches when assessing environmental impacts on marine ecosystems. Understanding interactions between pollutants and environmental stressors is crucial for effective conservation strategies. Future research should delve deeper into the impacts at lower biological levels and explore adaptive mechanisms in marine organisms facing multiple stressors. By doing so, we can better anticipate and mitigate the adverse effects of anthropogenic pollutants on marine biodiversity and ecosystem health., 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 Ltd.. All rights reserved.)

The production and consumption of plastic products had been steadily increasing over the years, leading to more plastic waste entering the environment. Plastic pollution is ubiquitous and comes in many types and forms. To enhance or modify their properties, chemical additives are added to plastic items during manufacturing. The presence and leakage of these additives, from managed and mismanaged plastic waste, into the environment are of growing concern. In this study, we gauged, via an online questionnaire, expert knowledge on the use, characteristics, monitoring and risks of plastic additives to the marine environment. We analysed the survey results against actual data to identify and prioritise risks and gaps. Participants also highlighted key factors for future consideration, including gaining a deeper understanding of the use and types of plastic additives, how they leach throughout the entire lifecycle, their toxicity, and the safety of alternative options. More extensive chemical regulation and an evaluation of the essentiality of their use should also be considered., 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 Ltd.. All rights reserved.)

The objective of this study was to determine whether and to what extent microplastics (MPs) enhance the toxicity of pollutants as well as whether pollutant-loaded MPs act as relevant vectors of chemical pollutants. With this aim, the toxicity for mussel and sea urchin embryos of: 1) three dissolved pollutants (Pol): chlorpyrifos (CPF), fluoranthene (FLT) and mercury (Hg); 2) their mixture with Microplastics (MP + Pol); and 3) pollutant-loaded MPs (MP Pol ), was assessed. Analyses of CPF, FLT and Hg were also performed to evaluate the transfer among dissolved and particulate phases. In general, the 'MP + Pol' treatments were more toxic as 48-h EC 50 (μg/L) than the 'Pol' treatments for sea urchin or mussel. The 48-h and 120-h EC 50 s (μg/L) for sea urchin showed little variation for CPF and MP + CPF, and no clear pattern was found for FLT and MP + FLT. The performed chemical analysis in the MP Pol tests indicated that desorption was the main route to explain the observed toxicity of Hg and a relevant route for CPF and FLT. This study contributes to improve the knowledge about the interactions between MPs and chemical pollutants, which is fundamental for a more realistic ecological risk assessment in aquatic ecosystems., 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 Ltd.. All rights reserved.)

The goal of the present work is to study whether ocean- acidification (OA) and -warming (OW) could increase the toxicity of pollutants on P. lividus. We studied how model pollutants such as chlorpyrifos (CPF) and microplastics (MP), alone or in combination, impact the fertilisation process, and the development of larvae under conditions of OA (dissolved inorganic carbon increase of 126 × 10 -6  mol per kg of sea water) and OW (temperature increase of 4 °C) predicted by FAO (Food and Agriculture Organization) for the next 50 years. Fertilisation was determined by microscopic examination after 1 h. Growth, morphology, and alteration level were measured after 48 h of incubation. Results showed that CPF has a marked effect on the growth of larvae, but less on the fertilisation rate. When larvae are exposed to both MP and CPF, the effect on fertilisation and growth is higher than when CPF is added alone. Larvae exposed to CPF tend to adopt a rounded shape which is detrimental to their buoyancy and the combination with other stressors aggravate this situation. The variables most influenced by CPF or its mixtures are those related to body length, body width, and higher levels of body abnormalities, which is consistent with the degenerative effects caused by CPF on sea urchin larvae. The PCA analysis showed that temperature has more influence when embryos or larvae are exposed to a combination of stressors, demonstrating that global climate change drastically increase the impact of CPF on aquatic ecosystems. Overall, in this work we demonstrated that global climate change conditions increase the sensitivity of embryos to MP and CPF. Our findings support the idea that global change conditions could have a severe impact on marine life, increasing the negative effect of toxic agents commonly present in the sea and their mixtures., 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. Published by Elsevier B.V.)

The growing use of plastics, including microplastics (MPs), has enhanced their potential release into aquatic environments, where microalgae represent the basis of food webs. Due to their physicochemical properties, MPs may act as carriers of organic and inorganic pollutants. The present study aimed to determine the toxicity of polyethylene MPs (plain and oxidized) and the model pollutants chlorpyrifos (CPF) and mercury (Hg) on the red microalgae Rhodomonas lens, to contribute to the understanding of the effects of MPs and associated pollutants on marine ecosystems, including the role of MPs as vectors of potentially harmful pollutants to marine food webs. R. lens cultures were exposed to MPs (1-1000 μg/L; 25-24,750 particles/mL), CPF (1-4900 μg/L), Hg (1-500 μg/L), and to CPF- and Hg-loaded MPs, for 96 h. Average specific growth rate (ASGR, day -1 ), cellular viability and pigment concentration (chlorophyll a, c2 and carotenoids) were measured at 48 and 96 h. No significant effects were observed on the growth pattern of the microalgae after 96-h exposure to plain and oxidized MPs. However, a significant increase in cell concentration was detected after 48-h exposure to plain MPs. A decrease of the ASGR was noticed after exposure to CPF, Hg and to CPF/Hg-loaded MPs, whereas viability was affected by exposure to MPs, CPF and Hg, alone and in combination. Chlorophyll a and c2 significantly decreased when microalgae were exposed to plain MPs and CPF, while both pigments significantly increased when exposed to CPF-loaded MPs. Similarly, chlorophyll and carotenoids content significantly decreased after exposure to Hg, whereas a significant increase in chlorophyll a was observed after 48-h exposure to Hg-loaded MPs, at the higher tested concentration. Overall, the presence of MPs modulates the toxicity of Hg and CPF to these microalgae, decreasing the toxic effects on R. lens, probably due to a lower bioavailability of the contaminants., 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 © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

The knowledge regarding the neurological and behavioral toxic effects associated with microplastics (MPs) and heavy metals exposure is still scarce. The present study aimed to evaluate the potential chronic (30 days) toxic effects of MPs (2 mg/L) and copper (Cu, 25 µg/L), alone or combined, in the zebrafish (Danio rerio) brain antioxidant system, cell proliferation/death, cholinergic-, serotonergic- and dopaminergic pathways and, consequently, in locomotor, anxiety, and social behaviors. Our findings showed that MPs and Cu exposure modulated the antioxidant system of zebrafish brain, with superoxide dismutase (SOD) and glutathione reductase (GR) having higher activity in the Cu25 +MPs group, but glutathione peroxidase (GPx) being inhibited in MPs, Cu25 and Cu25 +MPs. Moreover, an increase in acetylcholinesterase (AChE) activity was observed in all exposed groups. When considering neurogenesis genes, a downregulation of proliferating cell nuclear antigen (pcna) was noticed in zebrafish exposed to the mixture treatment, while for dopaminergic system-related genes (th and slc6a3) an upregulation was observed in MPs, Cu25 and Cu25 +MPs groups. An increase in apoptosis-related genes expression (casp8, casp9 and casp3) was observed in the MPs exposed group. Changes in zebrafish behavior, particularly in mean speed, total distance moved, inactivity in the aquaria, and social/shoaling behavior was also observed in the MPs and Cu exposed groups. Overall, our results highlight the multiplicity of toxic effects of MPs, alone or combined with Cu, in zebrafish brain, namely apoptosis and alterations in adult neurogenesis, neurocircuits and, consequently, behavior., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

In this study, zebrafish embryos were exposed to microplastics (MPs, 2 mg/L) and copper (Cu, 60 and 125 μg/L), alone or combined, for 14 days, and the development of motor neurons was assessed through gene expression and immunohistochemistry. DNA methyltransferases (DNMTs) genes expression was also evaluated. The results showed a downregulation of neuronal proliferation (sox2, pcna), neurogenesis (neuroD, olig2), and motor neurons development (islet) related genes, implying potential deficits in the neurogenesis of the exposed zebrafish early life stages. Downregulation of the maintenance and de novo DNMTs expression was also found, indicating that the DNA methylation patterns could be modulated by MPs and Cu. A high relative volume of proliferating cell nuclear antigen (PCNA)-positive cells was found in the fish retina from the MPs exposed group, suggesting that MPs increased the rate of cellular division. In contrast, a significant decrease of PCNA-positive cells, and therefore a lower cell proliferation, was found in the retina and brain of zebrafish exposed to Cu and Cu + MPs, which could lead to cognitive and behavioral functions impairment. No alterations were found in the relative volume of ISL1&2-positive cells. This study contributes to the knowledge of the mechanisms by which MPs and Cu cause neurotoxicity, fundamental for a comprehensive and realistic ecological risk assessment in aquatic populations., (Copyright © 2022 Elsevier B.V. All rights reserved.)

One of the major consequences of increasing atmospheric CO 2 is a phenomenon known as ocean acidification. This alteration of water chemistry can modulate the impact on marine organisms of other stressors also present in the environment, such as microplastics (MP). The objective of this work was to determine the combined impact of microplastic pollution and ocean acidification on the early development of Paracentrotus lividus. To study these multi-stressor impacts on development P. lividus the sea urchin embryo test (SET) was used. Newly fertilised embryos of P. lividus were exposed to a control treatment (filtered natural seawater), MP (3000 particles/mL), acidified sea water (pH = 7.6), and a combination of MP and acidification (3000 particles/mL + pH = 7.6). After 48, 72, and 96 h measurements of growth and morphometric parameters were taken. Results showed that ocean acidification and MP cause alterations in growth and larval morphology both before and after the larvae start to feed exogenously. The exposure to MP under conditions of ocean acidification did not produce any additional effect on growth, but differences were observed at the morphological level related to a decrease in the width of larvae at 48 h. Overall, changes in larvae shape observed at three key points of their development could modify their buoyancy affecting their ability to obtain and ingest food. Therefore, ocean acidification and MP pollution might compromise the chances of P. lividus to survive in the environment under future scenarios of global climate change., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Fish gills are in direct contact with the surrounding pollutants, and thus, potentially more vulnerable to microplastics (MPs) and heavy metals. The present study aimed to evaluate the long-term exposure effects of MPs and copper (Cu) in the gills of adult zebrafish (Danio rerio). To this end, zebrafish were exposed to MPs (2 mg/L), Cu (Cu25, 25 μg/L) and their mixture (Cu25 + MPs) for 30 days, and then oxidative stress, detoxification, antioxidant, metabolic and neurotoxicity enzymes/genes, as well serotonergic system and apoptosis genes, were evaluated in gills. In the mixture group, ROS levels were increased, while CAT and GPx activities were inhibited, indicating the induction of oxidative stress in zebrafish gills. This was followed by an increase of LPO levels and potential oxidative damage in zebrafish gills. The tryptophan hydroxylase 1a (tph1a) and caspase-3 (casp3) genes were significantly upregulated in Cu25 + MPs group, indicating a potential dysregulation of serotonin synthesis and apoptosis pathways, respectively. Overall, the present study contributes to improving the knowledge about the response of aquatic organisms to MPs and the potential ecological risk that these particles represent to the ecosystems., (Copyright © 2022 Elsevier Inc. All rights reserved.)

The effects of three relevant organic pollutants: chlorpyrifos (CPF), a widely used insecticide, triphenyl phosphate (TPHP), employed as flame retardant and as plastic additive, and bisphenol A (BPA), used primarily as plastic additive, on sea urchin (Paracentrotus lividus) larvae, were investigated. Experiments consisted of exposing sea urchin fertilized eggs throughout their development to the 4-arm pluteus larval stage. The antioxidant enzymes glutathione reductase (GR) and catalase (CAT), the phase II detoxification enzyme glutathione S-transferase (GST), and the neurotransmitter catabolism enzyme acetylcholinesterase (AChE) were assessed in combination with responses at the individual level (larval growth). CPF was the most toxic compound with 10 and 50% effective concentrations (EC 10 and EC 50 ) values of 60 and 279 μg/l (0.17 and 0.80 μM), followed by TPHP with EC 10 and EC 50 values of 224 and 1213 μg/l (0.68 and 3.7 μM), and by BPA with EC 10 and EC 50 values of 885 and 1549 μg/l (3.9 and 6.8 μM). The toxicity of the three compounds was attributed to oxidative stress, to the modulation of the AChE response, and/or to the reduction of the detoxification efficacy. Increasing trends in CAT activity were observed for BPA and, to a lower extent, for CPF. GR activity showed a bell-shaped response in larvae exposed to CPF, whereas BPA caused an increasing trend in GR. GST also displayed a bell-shaped response to CPF exposure and a decreasing trend was observed for TPHP. An inhibition pattern in AChE activity was observed at increasing BPA concentrations. A potential role of the GST in the metabolism of CPF was proposed, but not for TPHP or BPA, and a significant increase of AChE activity associated with oxidative stress was observed in TPHP-exposed larvae. Among the biochemical responses, the GR activity was found to be a reliable biomarker of exposure for sea urchin early-life stages, providing a first sign of damage. These results show that the integration of responses at the biochemical level with fitness-related responses (e.g., growth) may help to improve knowledge about the impact of toxic substances on marine ecosystems., (© 2022. The Author(s).)

Microplastics (MPs) are a major concern for marine ecosystems since they can be ingested by a wide range of marine species and transmitted through the food web. However, the potential hazardous impact of MPs in fishes, especially in early developing stages, is relatively unknown. In the present study, we assessed for the first time the ingestion and retention of MPs in early developing seahorses Hippocampus reidi. Seahorses are vulnerable species that may also be affected by both the direct ingestion of MPs through their preys and the accidental ingestion of MPs particles present in the water (i.e., seahorses ingest the prey by suction). We used copepods as both preys for seahorse juveniles and transfer vectors of MPs. Fed or starved copepods previously exposed to polyethylene microspheres (1-5 μm in diameter; 10 and 100 μg L-1) for 60 min at 26 °C showed fast evacuation of microspheres. The presence of MPs in copepods was significantly higher in previously fasted copepods compared to fed copepods. Seahorse juveniles fed on copepods pre-exposed to MPs, accumulated MPs in the gut proportionally to the concentration of MPs in copepods. A lower concentration of MPs in seahorses was observed at the longer exposure time (60 min), especially in fish fed with fasted copepods. However, after longer exposure, MPs were mainly accumulated close to the anus both individually or forming aggregates. Further studies should be performed to assess secondary effects of MPs ingestion in seahorses since they are considered a flagship species for marine conservation., 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.)

Plastics pose serious risks for fish productivity and a potential constraint for food security. Newly hatched blackspot seabream larvae were exposed to microplastics (MPs), copper (Cu, 10-810 µg/L) and their mixtures (Cu+MPs), during 3 and 9 days. Biochemical biomarkers and the expression of antioxidant and neurotoxicity-related genes were evaluated. In the 3-day exposure, catalase and glutathione-S-transferase activities decreased in MPs, Cu and Cu+MPs groups, followed by an increase of lipid peroxidation in the Cu270 and Cu270 +MPs exposed larvae. In the 9-day exposure, ROS levels increased in MPs and Cu30 groups, but no significant oxidative damage was observed, suggesting that the antioxidant system overcome the induced oxidative stress. However, the acetylcholinesterase transcript was downregulated in MPs, Cu and Cu10+MPs groups, indicating that MPs effects in cholinergic neurotransmission may arise after longer exposures. Overall, MPs and Cu can reduce survival, induce oxidative stress, lipid peroxidation, neurotoxicity, and impact negatively fish larvae fitness., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The Gulf of Cádiz is an area historically affected by acid mine drainage and also by the discharge of two important rivers, the Guadiana and the Guadalquivir. Metal and metalloid (As, Cd, Hg, Pb, Zn, Cu, Cr, Ni, Mn, Fe, Al and Li) concentrations were measured in 33 surface sediment samples, collected in the continental shelf, inside the Ría de Huelva and the Bay of Cádiz, to assess the metal pollution status of this area. Geographical distribution was identified for different groups of metals: Cu, Cd, Zn, Hg and As were mainly associated with discharges of the Tinto and Odiel rivers, which have been historically affected by mining, into the Ría de Huelva. Sediments inside this Ría presented the highest concentrations of the whole area for these metals, and enrichment factors revealed that Hg and Cu pollution spread up to 20 km away from the mouth of this Ría, into the continental shelf. On the contrary, the distribution of Ni, Cr and Mn did not show a clear gradient with distance from a pollution source, and was mainly explained by geochemical factors, such as their association with fine materials. The comparison of metal concentrations with sediment quality guidelines, predicts that adverse ecological effects due to the metal pollution may be occurring in a group of stations located in the northwest corner of the studied area. Local background levels for metals in the Gulf of Cádiz were proposed and the information obtained will be useful to identify those areas where sampling must be intensified in ongoing Marine Strategy Framework Directive (MSFD) monitoring programs. These results corroborate that metal pollution is still a serious problem in the Gulf of Cádiz., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Microplastics (MP) are contaminants of concern per se, and also by their capacity to sorb dissolved chemicals from seawater, acting as vehicles for their transfer into marine organisms. Still, the role of MP as vehicles for contaminants and their associated toxicological effects have been poorly investigated. In this work we have compared the role of MP (high density polyethylene, HDPE, ≤22 μm) and of natural organic particles (microalgae, MA) as vehicle for chlorpyrifos (CPF), one of the most common pesticides found in river and coastal waters. We have compared the capacity of MP and MA to carry CPF. Then, the mussel Mytilus galloprovincialis has been exposed for 21 days to dissolved CPF, and to the same amount of CPF loaded onto MP and MA. The concentration of CPF in mussel' tissues and several physiological, energetics and immune parameters have been analyzed after 7 and 21 days of exposure. Results showed similar CPF accumulation in mussel exposed to MP and to MA spiked with CPF. This revealed that MP acted as vector for CPF in a similar way (or even to a lesser extent) than MA. After 21 days of exposure mussels exposed to MP spiked with CPF displayed similar or more pronounced biological effects than mussels exposed to dissolved CPF or to MA loaded with CPF. This suggested that the combined "particle" and "organic contaminant" effect produced an alteration on the biological responses greater than that produced by each stressor alone, although this was evident only after 3 weeks of exposure., 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.)

The aim of this work was to estimate the potential risk of the combined effect of global change factors (acidification, temperature increase) and microplastic (MP) pollution on the growth and development of the sea urchin P. lividus. Embryo-larval bioassays were conducted to determine growth and morphology after 48 h of incubation with MP (1000 and 3000 particles/mL); with filtered sea water at pH = 7.6; and with their combinations. A second experiment was conducted to study the effect of pH and MP in combination with a temperature increase of 4 °C compared to control (20 °C). We found that the inhibition of growth in embryos reared at pH = 7.6 was around 75%. Larvae incubated at 3000 MP particles/mL showed a 20% decrease in growth compared to controls. The exposure to MP also induced an increase in the postoral arm separation or rounded vertices. The combined exposure to a pH 7.6 and MP caused a significant decrease of larval growth compared to control, to MP and to pH 7.6 treatments. Morphological alterations were observed in these treatments, including the development of only two arms. Increasing the temperature resulted in an increased growth in control, in pH 7.6 and pH 7.6 + MP3000 treatments, but the relative stomach volume decreased. However, when growth parameters were expressed per Degree-Days the lower growth provoked by the thermal stress was evidenced in all treatments. In this work we demonstrated that MP could aggravate the effect of a decreased pH and that an increase in water temperature generated an additional stress on P. lividus larvae, manifested in a lower growth and an altered development. Therefore, the combined stress caused by ocean warming, ocean acidification, and microplastic pollution, could threaten sea urchin populations leading to a potential impact on coastal ecosystems., 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.)

The evaluation of the interaction between microplastics (MPs) and heavy metals is of special importance for risk assessment. In this study, zebrafish (Danio rerio) were exposed to MPs (2 mg/L), two sub-lethal concentrations of copper (Cu, 60 and 125 μg/L) and their mixtures (Cu60 + MPs, Cu125 + MPs), from 2-h post-fertilization (hpf) until 14-days post-fertilization (dpf). Lethal and sublethal endpoints were evaluated, along with a set of biochemical and genetic biomarkers between 2 and 14 dpf. Exposure to MPs and Cu, single or combined, induced high mortality and oxidative stress in zebrafish larvae, with data showing that the antioxidant enzymes were inhibited at 6 dpf, increasing thereafter until 14 dpf, due to the accumulation of reactive oxygen species. MPs and Cu, single or combined, caused neurotoxicity in larvae by inhibiting acetylcholinesterase activity. There was an increased and significant effect of Cu + MPs groups on the evaluated biomarkers, concerning the corresponding Cu groups, suggesting that MPs may have a synergistic effect in relation to Cu. The Integrated Biomarker Response (IBR) evidenced that a higher degree of stress occurred at the larval period. Our findings highlight that MPs can act as a vector for heavy metals, therefore, influencing their bioavailability and toxicity in the organisms., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Microplastics (MPs, <5 mm) have been frequently detected in aquatic ecosystems, representing both health and ecological concerns. However data about the combined effects of MPs and other contaminants is still limited. This study aimed to evaluate the impact of MPs and the heavy metal copper (Cu) on zebrafish (Danio rerio) larvae development and behavior. Zebrafish embryos were subchronically exposed to MPs (2 mg/L), two sub-lethal concentrations of Cu (60 and 125 µg/L) and binary mixtures of MPs and Cu using the same concentrations, from 2-h post fertilization until 14 days post fertilization. Lethal and sub-lethal responses (mortality, hatching, body length) were evaluated during the embryogenesis period, and locomotor, avoidance, anxiety and shoaling behaviors, and acetylcholinesterase (AChE) activity were measured at 14 dpf. The results showed that survival of larvae was reduced in groups exposed to MPs, Cu and Cu+MPs. Regarding the behavioral patterns, the higher Cu concentration and mixtures decreased significantly the mean speed, the total distance traveled and the absolute turn angle, demonstrating an adverse effect on swimming competence of zebrafish larvae. Exposure to MPs and Cu, alone or combined, also affected avoidance behavior of zebrafish, with larvae not reacting to the aversive stimulus. There was a significant inhibition of AChE activity in larvae exposed to all experimental groups, compared to the control group. Moreover, a higher inhibition of AChE was noticed in larvae exposed to MPs and both Cu+MPs groups, comparatively to the Cu alone groups. Our findings demonstrate the adverse effects of MPs, alone or co-exposed with Cu, on fish early life stages behavior. This study highlights that MPs and heavy metals may have significant impacts on fish population fitness by disrupting locomotor and avoidance behaviors., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Data about the toxicological interactions of MPs and heavy metals in biota is limited, particularly in fish early life stages. This study aimed to evaluate the toxicological effects of MPs and copper (Cu), alone or combined, in zebrafish early life stages. Embryos were exposed from 2 until 96-h post-fertilization (hpf) to MPs (2 mg/L), three sub-lethal concentrations of Cu (15, 60 and 125 μg/L) and binary mixtures containing Cu and MPs (Cu15+MPs, Cu60+MPs, Cu125+MPs). Lethal and sub-lethal parameters, histopathological changes, biochemical biomarkers, gene expression and behavior were assessed. Our findings showed that Cu and Cu + MPs decreased embryos survival and hatching rate. Increased ROS levels were observed in larvae exposed to the two lowest Cu and Cu + MPs groups, suggesting an induction of oxidative stress. An increased CAT and GPx activities were observed in Cu and Cu + MPs, implying a response of the antioxidant defense system to overcome the metal and MPs stress. The sod1 expression was downregulated in all Cu groups and in the two highest Cu + MPs exposed groups. AChE was significantly inhibited in Cu and Cu + MPs groups, indicating neurotoxicity. A disruption of avoidance and social behaviors were also noticed in the Cu125 and Cu125+MPs exposed larvae. Evidences of Cu-toxicity modulation by MPs were observed in some endpoints. Overall, the findings of this study highlight that Cu alone or co-exposed with MPs lead to oxidative stress, neurotoxicity and ultimately behavioral alterations in early life stages of zebrafish, while MPs alone do not produce significant effects on zebrafish larvae., 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.)

Ten global harbours were assessed for sediment quality by quantifying the magnitude of anthropogenic change and ecological risk. Anthropogenic change (enrichment) was high for Derwent River and Sydney estuary, moderate for Santander Harbour, Rio de Janeiro and Dublin Port, slight for Hong Kong, minimal for Darwin. All 10 enrichment indices used showed similar results. Derwent River sediment was rated at high ecological risk, followed by Sydney and Santander estuaries with moderate risk. Auckland and Darwin sediments exhibited minimal ecological risk and sediment in the remaining harbours (Dublin, Hong Kong, Ravenna, Ria de Vigo and Rio de Janeiro) were assessed at slight ecological risk. The extraordinary variety of environments and types/quantities/qualities of data investigated resulted in as much a critique and development of methodology, as an assessment of human impact, including unique techniques for elemental normalisation and contaminant classification. Recommendations for an improved technical framework for sediment quality assessment are provided., Competing Interests: Declaration of competing interest The paper is an original manuscript that has not been published previously and is not currently being considered for publication elsewhere. The work is unfunded and there is no conflict of interest., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Ingestion of microplastics by marine organisms has been well documented, but their interaction with chemical pollutants has not been sufficiently addressed. The aim of this study was to determine the individual and combined effects of chlorpyrifos (CPF) and polyethylene microplastics (MP) on the survival, fecundity, feeding and egg viability of Acartia tonsa, a calanoid copepod widely distributed in planktonic communities. The median lethal concentration obtained for CPF was higher (LC 50  = 1.34 μg/L) than for the combination with MP (LC 50  = 0.37 μg/L), or CPF-loaded MP (LC 50  = 0.26 μg/L). Significant effects were also observed for feeding and egg production (EC 50  = 0.77 and 1.07 μg/L for CPF, 0.03 and 0.05 μg/L for CPF combined with MP, 0.18 and 0.20 μg/L for CPF-loaded MP). No significant effects were observed in the exposure to 'virgin' MP. This study confirms the role of MP as vectors of pollutants to marine organisms and supports the increased availability of certain toxicants carried out by MP. The effects observed in fitness-related responses suggest potential damage to A. tonsa populations. The comparison of the results obtained here with environmental concentrations indicates that the combined exposure to CPF and MP could constitute a risk to A. tonsa in the natural environment., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Biomarkers are useful tools to assess biological effects of pollutants that are extensively used in monitoring programs to assess ecosystem health. However, they are strongly affected by mussel physiological state, especially nutritive status, which has led to the search of new biological indicators of chemical pollutants exposition. Environmental metabolomics is an approach for examining the metabolic responses (measurement of low molecular weight endogenous metabolites) of an organism to both natural and anthropogenic stressors that can occur in its environment. The aim of the present work was to assess the effect of the polycyclic aromatic hydrocarbon fluoranthene (FLU) exposure on the metabolomic profiles of mussel digestive glands under different nutritive conditions. To achieve this objective, mussels were reared, for a period of 56 days, under three different food rations in order to obtain a gradient of nutritive status (negative, zero and positive energy balance), and after that, they were exposed, during 3 weeks, to a nominal concentration of 3 μg FLU L -1 . A total of 43 metabolites, including aminoacids (Ala, Val, Leu, Ile, etc.), energy metabolism related metabolites (ATP, AMP, etc.), organic osmolytes (taurine, etc.), redox metabolism (GSH, NADP+) and nucleotides, were identified and quantified in the digestive glands of the mussels. Principal Component Analysis (PCA) defined two principal components (PC1 and PC2) that explained 55.6% of the total variance, although the first component explains more than 80% of this variance, this being related to the mussel nutritive condition. The effect of the toxicant, explained by the PC2, is similar to that produced under conditions of food restriction, which masks the effect of the toxicant under these conditions. As the feeding conditions are more favorable, the toxic effect becomes more apparent. Therefore, the great influence of nutritive condition on mussel metabolome implies a handicap for the use of metabolomic biomarkers, as previously demonstrated for biochemical and other molecular biomarkers, in large-scale monitoring programs in which several food conditions coexist with pollution levels., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

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.)

Imposex is a disorder caused by organotins, mainly tributyltin, which results in the appearance of male sexual characteristics in females of gastropod mollusks. The main objective of this work was to make a critical analysis of the relationship between imposex and butyltin body burdens in Nucella lapillus and Nassarius reticulatus. Specifically, this study evaluates possible additive effects among butyltins, proposes scales of effects based on robust statistical criteria as alternatives to existing ones and defines the body burdens of TBT in N. lapillus and N. reticulatus corresponding to the assessment classes (ACs) of the Vas Deferens Sequence Index (VDSI) established by OSPAR. Data of organotin body burdens and biological effects was retrieved from the ICES Dataset and from scientific literature. All responses, except the percentage of females displaying Imposex (IMPF) in Nucella lapillus, showed a sigmoidal profile regarding to the body burden of mono- (MBT), di- (DBT) and tributyltin and sum of butyltins (SumBTs). TBT and the SumBTs were better indicators of the VDSI or Relative Penis Size Index/Relative Penis Length Index (RPSI/RPLI) responses than MBT or DBT in most cases. From a statistical point of view, RPSI/RPLI and VDSI were better indicators of contamination by TBT than IMPF, although both RPSI and RPLI showed lower sensitivity than VDSI. The model used for describing the joint effect of butyltins provided a statistically significant fitting to the data assuming a null effect for both MBT and DBT for N. lapillus, and a lower toxic contribution of MBT and DBT with respect to TBT for N. reticulatus. RPSI or RPLI values, equivalent to the ACs for VDSI, were proposed as alternative criteria when measuring moderate to high levels of imposex. TBT concentrations in N. reticulatus and N. lapillus tissues, corresponding to the ACs were calculated and provided valuable information for cross-species comparisons., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Legislation in the European Union (EU) aimed at reaching by 2015 a Good Ecological Status in regard to tributyltin (TBT, the biocide used in traditional antifouling paints). With a view to check such an achievement in N Atlantic Spain, baseline monitoring of gastropod imposex (the recommended assessment tool) was extended up to that date. In Galicia (the Western part of the study area) the use of the rock snail Nucella lapillus since 1996 had shown this environmental objective to be met as soon as 2009, but new surveys reveal no further improvement thereafter. As for the Eastern Cantabrian coast, imposex levels in the mud snail Nassarius reticulatus progressively declined from 2006 to 2015, when records finally complied with expectations. Both data sets are confronted and discussed in relation to the diverse environmental factors that may be determining the distribution of gastropods in these regions., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The need to create a new approach to carbon capture processes that are economically viable has led to the design and synthesis of sorbents that selectively capture carbon dioxide by physisorption. Solid Ionic Liquids (SoILs) were targeted because of their tunable properties and solid form under operational conditions. Molecular modelling was used to identify candidate SoILs and a number of materials based on the low cost, environmentally friendly acetate anion were selected. The materials showed excellent selectivity for carbon dioxide over nitrogen and oxygen and moderate sorption capacity. However, the rate of capture was extremely fast, in the order of a few seconds for a complete adsorb-desorb cycle, under pressure swing conditions from 1 to 10 bar. This showed the importance of rate of sorption cycling over capacity and demonstrates that smaller inventories of sorbents and smaller process equipment are required to capture low concentration CO 2 streams. Concentrated CO 2 was isolated by releasing the pressure back to atmospheric. The low volatility and thermal stability of SoILs mean that both plant costs and materials costs can be reduced and plant size considerably reduced.

Microplastic pollution has received increased attention over the last few years. This study documents microplastic ingestion in three commercially relevant demersal fish species from the Spanish Atlantic and Mediterranean coasts, the lesser spotted dogfish Scyliorhinus canicula, the European hake Merluccius merluccius and the red mullet Mullus barbatus. Overall 212 fish were examined, 72 dogfish, 12 hakes and 128 red mullets. The percentage of fish with microplastics was 17.5% (15.3% dogfish, 18.8% red mullets and 16.7% hakes), averaging 1.56±0.5 items per fish, and the size of the microplastics ranged from 0.38 to 3.1mm. These fish species are used currently as biomonitors for marine pollution monitoring within the Spanish Marine Pollution Monitoring Programme (SMP), and may be as well suitable candidates for monitoring spatial and temporal trends of ingested litter. The data presented here represent a baseline for the implementation of the Marine Strategy Framework Directive descriptor 10 in Spain., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The effect of the quality of two microalgal species on select biological and biochemical responses used as indicators of pollution were assessed. Mussels were conditioned for 6 weeks with the diatom Chaetoceros neogracile and the dinoflagellate Heterocapsa triquetra, chosen for being two clearly different types of primary production quality that differ in both biometric and biochemical characteristics. After dietary conditioning, the mussels were exposed to a polycyclic aromatic hydrocarbon, fluoranthene (FLU), for 1 week followed by 1 week of depuration. Results showed higher FLU accumulation in mussels fed on C. neogracile compared to those fed on H. triquetra. Concomitantly, a greater impact of this toxicant was observed in the biomarker responses of mussels fed on C. neogracile. These mussels showed an increase in the percentage of dead hemocytes, an activation of phagocytosis and ROS production of hemocytes after exposure. Some enzymatic activities also increased upon FLU exposure (superoxide dismutase -SOD-, catalase -CAT-, and glutathione reductases -GR-) and after depuration (glutathione-s-transferase -GST-). Results suggest that FLU exposure as well as food quality influence biomarker responses, with higher values of SOD, CAT and GR in non-exposed mussels fed on C. neogracile. In addition, upon exposure to the same FLU concentration, GR response varied according to dietary conditioning, suggesting that diet could act as a confounding factor in biomarker responses to pollution. Consequently, trophic conditions should be considered in marine pollution monitoring programs for a better interpretation of biomarker responses., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Biomarkers are useful tools to assess biological effects of pollutants and have been extensively used in monitoring programs to determine ecosystem health. In these programs, a wide range of environmental conditions are covered and sometimes, obtained data are difficult to interpret because of natural variables are affecting biomarker responses. Among these variables, musseĺs reproductive status has been considered one of the most changing variables between sites in a monitoring survey. Thus, the main aim of this work was to identify the effect that mussel reproductive status has on biomarker responses. For that purpose, mussels sampled at two periods in the reproductive cycle (reproductive and resting stages) were conditioned to the same laboratory conditions and exposed to fluoranthene (FLU) for three weeks. Studied biomarkers covering a wide range of organism responses were included: bioaccumulation, physiological rates (clearance rate -CR-, absorption efficiency -AE-, respiration rate -RR- and their integration in the scope for growth -SFG-), antioxidant enzyme activities (superoxide-dismutase -SOD-, catalase -CAT-, glutathione reductase -GR-, glutathione peroxidase -GPx-, glutathione-S-tranferase -GST-) and biochemical damage responses (lipid membrane peroxidation -LPO-). The results obtained evidenced that the levels of the biomarkers studied (RR, SOD, CAT and GPx) were higher at reproductive than at resting stage. On the other hand, the effect of toxicant was observed in SFG, CAT and GPx but this effect was only detected during the resting period. Moreover, there was a deterioration of mussel gonadal tissue with FLU exposure during reproductive stage. FLU accumulation in mussel tissues was also dependent of the reproductive status with higher internal concentrations during resting than reproductive period. In conclusion, there was a strong effect of reproductive status on studied biomarkers which seems to mask the effect of FLU at reproductive stage. The present study evidences the need to include the measurement of mussel biological status in marine pollution monitoring programs for a correct interpretation of biomarker data., (Copyright © 2016 Elsevier B.V. All rights reserved.)

In this study, PCB-153 bioaccumulation kinetics and concentration-response experiments were performed employing wild Mytilus galloprovincialis mussels. In addition, the activity of three enzymatic biomarkers: glutathione S-transferase (GST), glutathione peroxidase (GPx) and acetylcholinesterase (AChE), were measured in the mussel gills. The experimental data fitted well to an asymptotic accumulation model with a high bioconcentration factor (BCF) of 9324 L kg(-1) and a very limited depuration capacity, described by a low excretion rate coefficient (Kd = 0.083 d(-1)). This study reports by first time in mussels significant inhibition of GST activity and significant induction of GPx activity as a result of exposure to dissolved PCB-153. In contrast, AChE activity was unaffected at all concentrations and exposure times tested. The effects on both enzymes are time-dependent, which stresses the difficulties inherent to the use of these biomarkers in chemical pollution monitoring programs., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Guano is an important source of marine-derived nutrients to seabird nesting areas. Seabirds usually present high levels of metals and other contaminants because the bioaccumulation processes and biotic depositions can increase the concentration of pollutants in the receiving environments. The objectives of this study were to investigate: the toxicity of seabird guano and the joint toxicity of guano, Cu and Pb by using the sea urchin embryo-larval bioassay. In a first experiment, aqueous extracts of guano were prepared at two loading rates (0.462 and 1.952 g L(-1)) and toxicity to sea-urchin embryos was tested. Toxicity was low and not dependent of the load of guano used (EC50 0.42 ± 0.03 g L(-1)). Trace metal concentrations were also low either in guano or in aqueous extracts of guano and the toxicity of extracts were apparently related to dissolved organic matter. In a second experiment, the toxicity of Cu-Pb mixtures in artificial seawater and in extracts of guano (at two loadings: 0.015 and 0.073 g L(-1)), was tested. According to individual fittings, Cu added to extracts of guano showed less toxicity than when dissolved in artificial seawater. The response surfaces obtained for mixtures of Cu and Pb in artificial seawater, and in 0.015 g L(-1) and 0.073 g L(-1) of guano, were better described by Independent Action model adapted to describe antagonism, than by the other proposed models. This implied accepting that EC50 for Cu and Pb increased with the load of guano and with a greater interaction for Cu than for Pb., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Biomarkers have been extensively used in monitoring programs with the aim of assessing the biological effects of pollutants on marine organisms and determining environmental status. Data obtained from these programs are sometimes difficult to interpret due to the large amount of natural variables affecting biological processes, which could act as confounding factors on biomarker responses. The main aim of this work was to identify the effect of one of these variables, the food availability, and consequently, the mussel nutritive status, on biomarker responses. For that purpose, mussels (Mytilus galloprovincialis) were conditioned to three different food rations for 2 months in order to create three mussel nutritive statuses and afterwards, each status was exposed to three nominal concentrations of fluoranthene (FLU) for 3 weeks. A battery of biomarkers was considered in this study to cover a wide range of organism responses, both physiological (scope for growth - SFG) and biochemical (superoxide dismutase - SOD, catalase - CAT, glutathione reductase - GR, glutathione peroxidase - GPx, glutathione-S-transferase - GST and phenoloxidase - PO activities, and lipid membrane peroxidation - LPO). The results obtained, evidenced that most of the studied biomarkers (SFG, SOD, CAT, GPx, and PO) were strongly affected by mussel nutritive status, showing higher values at lower status, whereas the effect of toxicant was not always evident, masked by the nutritive status effect. This paper demonstrates that toxicants are not the only source of variability modulating pollution biomarkers, and confirms nutritive status as a major factor altering biochemical and physiological biomarkers., (Copyright © 2015 Elsevier B.V. All rights reserved.)

We have established for first time an ecotoxicological bioassay using cryopreserved sea urchin embryos (Paracentotus lividus) and provided a comparison to the already standardized sea urchin embryo-larval bioassay, using selected (organic and inorganic) pollutants and sediment elutriates from 4 different locations from Ria de Vigo harbour (Galicia, NW Iberian Peninsula). A cryopreservation protocol was designed in order to enable the successful cryopreservation and cryobanking of gametes and embryos to be used for marine quality assessment and ensure the accessibility to high quality reproductive material all year round, as an option to conditioning adults for out of season reproduction. The calculated EC50 using the cryopreserved blastula was 53.7 μg L(-1) for copper, 81.0 μg L(-1) for lead, 300.6 μg L(-1) for BP-3 and 300.6 μg L(-1) for 4-MBC. The sensitivity of the classic sea urchin embryo-larval bioassay was compared with the bioassay conducted with cryopreserved blastula. The results showed that the use of cryopreserved blastula bioassay allows detecting lower concentrations of pollutants in comparison with the classic bioassay., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The anthropogenic emissions of Pt to the environment have increased significantly over the past decades, especially after the introduction of the catalytic converters in motor vehicles. In order to check whether this is affecting the levels of this trace metal on living organisms, time-series analysis of freeze-dried soft tissue material of wild mussels (Mytilus galloprovincialis) covering the period from 1991 to 2001 and collected at an urban beach in the city of Vigo (NW Iberian Peninsula) was conducted. Concentrations ranged from 0.30 to 0.68 ng g(-1) with an average concentration of 0.47 ± 0.10 ng g(-1) (n=21); these concentrations were higher than those obtained for samples collected at a control location away from anthropogenic pressure (0.31 ± 0.10 ng g(-1); n=5). Platinum concentrations followed a statistically significant temporal trend (at the 0.020 level), and the excess of Pt in mussels over the 1991-2011 period compared to the control location were correlated with the European Pt autocatalyst demand (p=0.0006) and, especially, the car sales in Spain (p=0.0001). A bioaccumulation factor of ~5·10(3) was derived, which is greater than those previously calculated for Pt from exposure experiments, but 1-2 orders of magnitude lower than other trace elements (e.g. Zn, Cu, Pb, Cd)., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Wild marine mussels, Mytilus galloprovincialis showed a moderate bioaccumulation ability when exposed to waterborne 4-nonylphenol (4-NP), with a bioconcentration factor (BCF) of 6850 L Kg(-1) (dry weight). Kinetic and concentration-response experiments were performed and three enzymatic biomarkers in mussel gills were measured: Glutathione S-transferase (GST), glutathione peroxidase (GPx) and acetylcholinesterase (AChE). Exposure of mussels to environmentally relevant concentrations (25-100 μg L(-1)) of 4-nonylphenol significantly inhibited the AChE activity and induced the GST and GPx activities. GST induction was dose dependent whilst GPx activity showed a less consistent pattern, but in both cases the induction remained after a 10 d depuration period. Mussels seem capable of eliminating 4-NP from their tissues through a mechanism involving GST induction., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Mussels, Mytilus galloprovincialis, showed a high bioaccumulation ability when exposed to waterborne tetrabromodiphenyl ether (BDE-47), with a bioconcentration factor of 10,900 L Kg(-1) wet weight, and slow depuration rates in clean seawater. Kinetic and concentration-response experiments were performed measuring in the exposed mussel the activities of three molecular biomarkers: glutathione S-transferase (GST), glutathione peroxidase (GPx) and acetylcholinesterase (AChE). The long term (30 days) exposure of mussels to all concentrations (2-15 µg L(-1)) of BDE-47 significantly inhibited the AChE and GST activities, a result that supports the suitability of these biomarkers in marine pollution monitoring programs. However, GPx activity showed a less consistent pattern of response depending on the concentration and the duration of exposure.

This study deals with the identification and characterization of biological variables that may affect some of the biological responses used as pollution biomarkers. With this aim, during the 2012 mussel survey of the Spanish Marine Pollution monitoring program (SMP), at the North-Atlantic coast, several quantitative and qualitative biological variables were measured (corporal and shell indices, gonadal development and reserves composition). Studied biomarkers were antioxidant enzymatic activities (CAT, GST, GR), lipid peroxidation (LPO) and the physiological rates integrated in the SFG biomarker (CR, AE, RR). Site pollution was considered as the chemical concentration in the whole tissues of mussels. A great geographical variability was observed for the biological variables, which was mainly linked to the differences in food availability along the studied region. An inverse relationship between antioxidant enzymes and the nutritional status of the organism was evidenced, whereas LPO was positively related to nutritional status and, therefore, with higher metabolic costs, with their associated ROS generation. Mussel condition was also inversely related to CR, and therefore to SFG, suggesting that mussels keep an "ecological memory" from the habitat where they have been collected. No overall relationship was observed between pollution and biomarkers, but a significant overall effect of biological variables on both biochemical and physiological biomarkers was evidenced. It was concluded that when a wide range of certain environmental factors, as food availability, coexist in the same monitoring program, it determines a great variability in mussel populations which mask the effect of contaminants on biomarkers., (Copyright © 2014 Elsevier Inc. All rights reserved.)

The aim of this note is to discuss the relevance of the interaction/integration of monitoring of contaminants for the protection of the marine environment and for human health safety (descriptors 8 and 9, respectively) within the Marine Strategy Framework Directive (MSFD). The identification of possible relations between contaminant levels in sediments and tissues of fish and other seafood, as well as the association of those levels to pollution sources, are major challenges for marine researchers. The Spanish initial assessment in the North-East Atlantic marine region was used as an example to show some gaps and loopholes when dealing with the relationship between descriptors 8 and 9. The main problem to deal with is that monitoring programmes intended for the assessment of marine environmental quality and for human health safety usually apply different approaches and methodologies, and even different tissues are analysed in some species (mainly fish). It is therefore recommended to make a profound revision of current sampling strategies, procedures and methodologies, including the selection of target species and tissues and to improve the traceability of samples of fish and other seafood for human consumption. On the other hand, despite the scope of descriptor 9 which is limited to commercially relevant species, this fact should not be an obstacle in the application of the 'ecosystem approach' within the MSFD. In order to appropriately solve these shortcomings, an information exchange system between authorities dealing with descriptors 8 and 9 should be strongly encouraged for the next steps of the MSFD's implementation.

Background: The cryopreservation of embryos can be a powerful biotechnological tool to supply all year-round biological material for sea urchin aquaculture production. This study investigates different methodological and biological factors that may affect the result of the cryopreservation process of sea urchin (Paracentrotus lividus) embryos. Our data indicate that neither embryo density nor the use of different cryopreservation containers presented effect on the cryopreservation outcome. Contrary to other marine invertebrates, for sea urchin embryo cryopreservation ultrapure water cannot be used as CPA solvent, yielding zero survival. After studying the reproductive parameters along the reproductive season, we found a positive correlation between both male and female Condition Index (C.I.), and between the oocyte weight and C.I. Both the histology study of female gonads and the C.I. variation, suggest that the sea urchin natural spawning period in the Ría de Vigo occurs between June and July. We found no correlation between any of the reproductive parameters monitored and the cryopreservation outcome.

This study undertakes an overall assessment of pollution in a large region (over 2500 km of coastline) of the N-NW Spanish coast, by combining the use of biochemical (AChE, GST, GPx) and physiological (SFG) responses to pollution, with chemical analyses in wild mussel populations (Mytilus galloprovincialis). The application of chemical analysis and biological techniques identified polluted sites and quantified the level of toxicity. High levels of pollutants were found in mussel populations located close to major cities and industrialized areas and, in general, average concentrations were higher in the Cantabrian than in the Iberian Atlantic coast. AChE activities ranged between 5.8 and 27.1 nmol/min/mg prot, showing inhibition in 12 sampling sites, according to available ecotoxicological criteria. GST activities ranged between 29.5 and 112.7 nmol/min/mg prot, and extreme variability was observed in GPx, showing activities between 2.6 and 64.5 nmol/min/mg prot. Regarding SFG, only 5 sites showed 'moderate stress' (SFG value below 20 J/g/h), and most sites presented a 'high potential growth' (>35 J/g/h) corresponding to a 'healthy state'. Multivariate statistical techniques applied to the chemical and biological data identified PCBs, organochlorine pesticides and BDEs as the main responsible of the observed toxicity. However, the alteration of biological responses caused by pollutants seems to be, in general, masked by biological variables, namely age and mussel condition, which have an effect on the mussels' response to pollutant exposure., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

As part of an integrative monitoring campaign involving water and sediment chemistry, in situ bioassays, and mussel bioaccumulation and biomarkers, Mytilus galloprovincialis mussels of standard size were transplanted from a clean location to five sites in two important harbours from the Atlantic coast of Spain (Vigo and Pasaia). After a 30-day field exposure, the concentrations of major contaminants (trace metals, polychlorinated biphenyls and polycyclic aromatic hydrocarbons) accumulated in mussel tissues were measured at each site, and a mussel bioaccumulation index (MBI) was calculated. The enzymatic activity levels of glutathione S-transferase (GST), glutathione peroxidase (GPx) and acetylcholinesterase (AChE) were quantified in the gills of transplanted mussels (n=12). Mussels from the most polluted sites consistently exhibited significantly higher GST and GPx activities compared to the control site, whereas AChE activity was significantly inhibited. The responses of the GST and GPx activities were related to MBI, trace metals and PAH concentrations in mussels, whereas AChE activity was related to the trace metals concentrations in mussels. The above results suggest that GST and AChE activities can be used as potential biomarkers for active monitoring in marine coastal ecosystems. However, at this moment, GPx activity is not robust enough to be applicable to harbour areas., (© 2013.)

Oysters and mussels are among the most farmed species in aquaculture industry around the world. The aim of this study was to test the toxicity of cryoprotective agents to trochophore larvae from two different species of bivalves and develop an improved cryopreservation protocol to ensure greater efficiency in the development of cryopreserved trochophores (14 h old oyster larvae and 20 h old mussel larvae) to normal D-larvae for future developments of hatchery spat production. The cryopreservation protocol producing the best results for oyster trochophores (60.0 ± 6.7% normal D-larvae) was obtained by holding at 0 °C for 5 min then cooling at 1 °C min(-1) to -10 °C and holding for 5 min before cooling at 0.5 °C to -35 °C, holding 5 min and then plunging into liquid nitrogen (LN), using 10% ethylene glycol. For mussel experiments, no significant differences were found when cooling at 0.5 °C min(-1) or at 1 °C min(-1) for CPA combinations with 10% ethylene glycol and at 0.5 °C min(-1). Using these combinations, around half of trochophores were able to develop to normal D-larvae post-thawing (48.9 ± 7.6% normal D-larvae)., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Biomarkers are required to assess the biological effects of pollutants on marine organisms in order to monitor ecosystem status, but their use is often limited by their strong variability due to environmental and/or intrinsic biological factors. Accordingly, the main aim of this work was to set up practical procedures for a battery of widely used biomarkers in mussels (Mytilus galloprovincialis). Antioxidant enzymes (catalase [CAT] and glutathione peroxidase [GPx]), a phase II detoxification enzyme (glutathione S-transferase [GST]) and a neurotransmitter catabolism enzyme (acetylcholinesterase [AChE]), were considered. Several relevant aspects were studied in order to obtain a more realistic interpretation of biomarker responses, including the calculation of the minimum sample size required to estimate the population mean with a fixed error margin, the selection of the specific organ or tissue where the enzymatic activity is higher for each biomarker, and the influence of tidal height and temperature on the basal enzymatic activity. GST and CAT activities needed a minimum sample size of 12, whereas for GPx and AChE activities a minimum sample size of 14 was required. The gills were the organ with higher GST, GPx and AChE enzymatic activities, whereas the digestive gland showed the highest CAT activity. Also, the low inter-tidal was the recommended tide level whilst no significant effect of temperature was observed on GST, GPx and CAT, and no clear pattern could be identified for AChE. The implications for environmental monitoring are discussed., (Copyright © 2013 Elsevier B.V. All rights reserved.)