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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.)

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.)

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.)

wWeathering of petroleum spilled in the marine environment may not only change its physical and chemical properties but also its effects on the marine ecosystem. The objective of this study was to evaluate the toxicity of the water-accommodated fraction (WAF) obtained from a standard fuel oil following an environmentally realistic simulated weathering process for a period of 80 d. Experimental flasks with 40 g L(-1) of fuel oil were incubated at 18°C with a 14 h light:10 h dark photoperiod and a photosynthetically active radiation (PAR) intensity of 70 μE m(-2) s(-1). Samples were taken at four weathering periods: 24 h, 7, 21 and 80 d. WAF toxicity was tested using the sea urchin (Paracentrotus lividus) and mussel (Mytilus galloprovincialis) embryo-larval bioassays and the aromatic hydrocarbons levels (AH) in the WAF were measured by gas chromatography/mass spectrometry. In contrast with the classic assumption of toxicity decrease with oil weathering, the present study shows a progressive increase in WAF toxicity with weathering, being the EC(50) after 80d eightfold lower than the EC(50) at day 1, whereas AH concentration slightly decreased. In the long term, inoculation of WAF with bacteria from a hydrocarbon chronically-polluted harbor slightly reduced toxicity. The differences in toxicity between fresh and weathered fuels could not be explained on the basis of the total AH content and the formation of oxidized derivatives is suggested to explain this toxicity increase., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Concentrations of polychlorinated biphenyls (PCBs) were determined in wild mussels (Mytilus galloprovincialis) from seven areas of the N-NW Spanish coast, during the period 1991-2009. The studied area is comprised of highly productive ecosystems which support important commercial fishing and shellfishing activities. The seven PCBs indicator congeners recommended by ICES (IUPAC nos. 28, 52, 101, 118, 138, 153 and 180), were analysed by gas chromatography with an electron capture detector (GC-ECD). Lowest average concentrations of PCBs were recorded in Arousa (1.04 μg kg(-1) wet weight), a low populated area with no industry; while the highest levels were found in areas near cities and/or showing a high industrial activity such as: Bilbao Ziérbena (139 μg kg(-1) ww) or A Coruña (55 μg kg(-1) ww). The hexachlorinated congeners CB153 and CB138 dominated the profiles in all wild mussel populations, although a geographical pattern was found in which the relative contributions of the lower chlorinated congeners were higher in the west coast (Rías of Vigo, Pontevedra and Arousa), which may reflect the occurrence of recent inputs of those compounds in the area. Finally, the application of the Mann-Kendall's test demonstrate the existence of a significant decrease in PCBs levels since 1991 up to 1996 in the studied area, whereas either a slow decrease or stable concentrations were registered over the past decade., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

In the present work, we investigated the potential use of several antioxidant enzymes in wild mussels (Mytilus galloprovincialis) as biomarkers of marine pollution. The enzymatic activity levels of glutathione S-transferase (GST), glutathione peroxidase (GPx) and catalase (CAT) were measured in gills and digestive gland. Those enzymes participate in the cellular defense system that is involved in the adaptive response of organisms to chemical pollution. Adult mussels were collected at five sampling sites located at the Ría de Pontevedra and Ría de Vigo. Seasonal variations in the control site were observed for the CAT activity, but no significant variability was found for GST and GPx. Mussels from the most polluted sites exhibited a significantly greater GST activity compared to the control site (p<0.05) during the sampling period, whereas GPx and CAT activities have not such a marked pattern. Trace metals, PAHs, PCBs and DDT contents in mussels at sampling sites showed highly significant positive correlations with the GST activity. This study supports the use of GST as a useful biomarker for long-term pollution monitoring in marine coastal ecosystems, whilst more research is needed for GPx and CAT., (Copyright 2009 Elsevier Ltd. All rights reserved.)

Elutriate embryo-larval bioassays with sea-urchins (Paracentrotus lividus) and ascidians (Ciona intestinalis) were conducted concurrently with trace metal analyses as part of an integrative assessment of sediment pollution at Ría de Pontevedra (Galicia, NW Spain). High metal contents in sediments were found in localised areas from the inner part of the estuary indicating a clear anthropogenic influence. In particular, very high Cu, Zn and Pb levels were found at sites P2 and P3, which were also the most toxic to the embryo-larval bioassays. Sediment quality guidelines were used to help in the ecological interpretation of sediment chemistry data and to identify pollutants of concern. Cu and Zn in P3 were consistently above the effects range median (ERM) values, which seem to be good predictors of toxicity to sea-urchin and ascidian embryos. A toxic unit approach, based on published EC(50) values and metal levels in elutriates, was used to assess the harmful ecological effects associated to sediment chemistry. Toxicity detected in P3 may be explained on the basis of the toxic unit model; however, the high toxicity detected at P2 may be attributable not only to the metals quantified in the analyses but also to unmeasured organic pollutants. Multidimensional scaling applied independently to the toxicology and chemistry data resulted in a good agreement between both types of configurations. Moreover, the Mantel test revealed a significant correlation (r(M)=0.481; p=0.019) between metal concentrations and toxicity data profiles, supporting the correspondence between configurations.

The effects of medetomidine, a novel antifouling candidate, on the burrowing bivalve Abra nitida were studied. The burrowing behaviour, sediment reworking activity and faeces production were assessed after 24 h exposure of A. nitida to sublethal concentrations of medetomidine. Medetomidine caused a significant decrease in the burrowing response and in the sediment reworking activity. The median effective concentrations (EC50) were 430 nM (86 microg/l) and 4.4 nM (0.9 microg/l), respectively. No effects on the faeces production were detected. Although significant effects of medetomidine on A. nitida were registered, a reversibility of the effects was observed when 24 h-exposed animals were incubated in clean seawater and sediment for 24 h. Considerations relating to the future commercialisation of medetomidine for antifouling purposes are discussed.