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Despite the recent focus on hazardous and noxious substances (HNS) spills preparedness and responses, much remains to be done regarding the threat posed by HNS spills on marine biota. Among the identified priority HNS,p-xylene was selected to conduct ecotoxicological assays. The aim of this study was to assess the performance of the amphipodGammarus locustaunder acute and chronic exposure top-xylene simulating conditions of a spill incident. In the acute exposure (96 h) thep-xylene LC50was estimated. In the chronic bioassay (36 d), an integration of organism-level endpoints (survival, growth rate, and sex ratio) with biochemical markers indicative of oxidative stress including catalase (CAT), glutathioneS-transferase (GST), and superoxide dismutase (SOD) activities and lipid peroxidation (LPO) levels was determined. The aim was to increase the xylene ecotoxicological database and better predict its impact in aquatic environments.p-Xylene induced several chronic toxicity effects inG. locusta. Significant alterations in antioxidant enzymes and lipid peroxidation levels as well as growth rate and biased sex-ratio were observed. p-Xylene significantly affected the activities of CAT, SOD, and GST inG. locustaand produced oxidative damage by increasing levels of LPO in males. Further, impacts in key ecological endpoints, that is, growth and sex ratio, were noted that might be indicative of potential effects at the population level in a spill scenario. The present data may be useful to assist relevant bodies in preparedness and response to HNS spills. [ABSTRACT FROM AUTHOR]

The nonsteroidal anti-inflammatory drug naproxen (NPX) is among the most consumed pharmaceuticals worldwide, being detected in surface waters within the ng to μg/L range. Considering the limited chronic ecotoxicity data available for NPX in aquatic ecosystems, the present study aimed at evaluating its impact in the model organism Danio rerio , following a full life-cycle exposure to environmentally relevant concentrations (0.1 to 5.0 μg/L). An integration of apical endpoints, i.e., survival, growth, and reproduction, with gonad histopathology and gene transcription (RNA-seq) was performed to provide additional insights into the mode of action (MoA) of NPX. NPX decreased zebrafish growth and reproduction and led to histopathological alterations in gonads at concentrations as low as 0.1 μg/L. At the molecular level, 0.7 μg/L of NPX led to a disruption in gonads transcription of genes involved in several biological processes associated with reproduction, mainly involving steroid hormone biosynthesis and epigenetic/epitranscriptomic machineries. Collectively, these results show that environmentally realistic concentrations of NPX affect zebrafish reproduction and associated signaling pathways, indicating that current hazard and risk assessment data for NPX underestimate the environmental risk of this pharmaceutical.

Introduction: When it comes to disease modeling, countless models are available for Lysosomal Storage Diseases (LSD). Historically, two major approaches are well-established: in vitro assessments are performed in patient fibroblasts, while in vivo pre-clinical studies are performed in mouse models. Still, both platforms have a series of drawbacks. Thus, we implemented two alternative and innovative protocols to mimic a particular sub-group of LSDs, the Mucopolysaccharidoses both in vitro and in vivo., Methods: The first one relies on a non-invasive approach using dental pulp stem cells from deciduous teeth (SHEDs). SHEDs are multipotent neuronal precursors that can easily be collected. The second uses a state-of-the-art gene editing technology (CRISPR/Cas9) to generate zebrafish disease models., Results: Even though this is an ongoing project, we have already established and characterized two MPS II and one MPS VI SHED cell models. These cells self-maintain through several passages and can give rise to a variety of cells including neurons. Furthermore, all MPS-associated sub-cellular phenotypes we have assessed so far are easily observable in these cells. Regarding our zebrafish models, we have successfully knocked down both naglu and hgsnat and the first results we got from the behavioral analysis are promising ones, as we can observe altered activity and sleep patterns in the genetically modified fish. For this particular approach we chose MPS III forms as our target disorders, since their neurological features (hyperactivity, seizures and motor impairment) and lifespan decrease would be easily recognizable in zebrafish., Conclusion: Now that these methods are well-established in our lab, their potential is immense. On one hand, the newly developed models will be of ultimate value to understand the mechanisms underlying MPS sub-cellular pathology, which have to be further elucidated. On the other hand, they will constitute an optimal platform for drug testing in house. Also noteworthy, our models will be published as lab resources and made available for the whole LSD community., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

With the growing interest to exploit mineral resources in the deep-sea, there is the need to establish guidelines and frameworks to support hazard and risk assessment schemes. The present study used a subtidal species of filter-feeding bivalve, the clam Spisula solida, as a proxy to better understand the impacts of sediment plumes in marine organisms under hyperbaric conditions. Four concentrations of suspended sediments (0 g/L, 1 g/L, 2 g/L, and 4 g/L) were used in a mixture with different grain sizes at 4 Bar for 96 h. Functional (filtration rate-FR) and biochemical endpoints (catalase-CAT, glutathione s-transferase-GST, and lipid peroxidation-LPO) were analyzed in the gonads, digestive gland, and gills of S. solida after a 96-h exposure at 4 Bar (the natural limit of the species vertical distribution). The FR showed a decreasing trend with the increasing sediment concentrations (significant effects at 2 and 4 g/L). Additionally, significant changes were observed for some of the tested oxidative stress biomarkers, which were concentration and tissue-dependent, i.e., CAT activity was significantly elevated in gills (1 g/L treatment), and GST was decreased in digestive gland (1 g/L treatment). Overall, the results show that suspended sediments, at 2 and 4 g/L, have negative functional impacts in the bivalve S. solida providing additional insights to improve hazard assessment of deep-sea mining. These findings represent a step forward to ensure the mitigation of the potential negative effects of deep-sea resource exploitation., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

A membrane ozone contactor, operated under continuous mode, was applied to promote the tertiary treatment of urban wastewater (UWW), targeting the removal of contaminants of emerging concern (CECs), bacterial disinfection, and toxicity reduction. This system relies on the homogeneous radial distribution of ozone (O 3 ) in the reaction zone by "titration" through a microfiltration borosilicate tubular membrane, while the UWW swirls around the membrane and drags the O 3 microbubbles generated in the membrane shell-side. The membrane is coated with titanium dioxide (TiO 2 -P25) and radiation can be externally supplied via four UV lamps. The ozonation tests were carried out with secondary-treated UWW collected in different seasons (winter and summer) and spiked with a mix of 19 CECs (10 μg L -1 each). For an O 3 dose of 18 g m -3 , the best performance was obtained by increasing the O 3 concentration (maximum [O 3 ] G,inlet of 200 g Nm -3 ) and decreasing the gas flow rate (minimum Q G of 0.15 Ndm 3 min -1 ), providing the highest ozone transfer yield (88 %) and, thus higher specific ozone dose (g O 3 per g dissolved organic carbon). Under these conditions, removals >80 % or concentrations below the limit of quantification were obtained for up to 13 of the 19 CECs and reductions up to 5 log units for total heterotrophs and below the limit of detection for enterobacteria and enterococci. Tests including a UVC dose of 0.10 kJ L -1 enhanced disinfection ability but had no impact on CECs oxidation. After ozonation, the abundance of antibiotic resistant bacteria was reduced but not eliminated, and microbial regrowth after 3-day storage was observed. No toxic effect was detected on zebrafish embryos using a dilution factor of 4 for the ozonized UWW and when granular activated carbon adsorption was subsequently applied the dilution factor decreased to 2., 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.)

This work investigated, during one year, the occurrence and fate of 52 contaminants of emerging concern (CECs) in transnational river basins and coastal areas of the North of Portugal and Galicia (NW Spain) and the wastewater treatment plants (WWTPs) discharging on these environments. The different CECs investigated included pharmaceuticals, personal care products, industrial chemicals, among others, of which ca. 90 % would fulfill the persistence, mobility and toxicity criteria proposed by the German Environmental Agency. The results showed the ubiquitous presence of these CECs and an incomplete removal of over 60 % of them with current conventional WWTPs. These findings highlight the requirement of a prominent and coordinated upgrade of WWTP treatments in order to meet the future European Union regulations on urban wastewater treatment and surface water quality. In fact, even some compounds exhibiting high removals, such as caffeine or xylene sulfonate, were frequently detected in river and estuarine waters at the high ng L -1 level. Thus, our preliminary risk assessment study concluded that 18 of the CECs presented a potential risk for the environment, being caffeine, sulpiride, perfluorooctanoic acid (PFOA), diclofenac, fipronil and perfluorobutanoic acid (PFBA) the most concerning ones. Yet, additional toxicity data as well as a more robust information on persistence and mobility of CECs are necessary for better estimating the magnitude of the problem and improve risk assessment. As an example, in the case of the antidiabetic metformin, recent research has revealed toxicity for model fish species at concentration levels below those found in 40 % of the river water samples analyzed in this work., 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.)

Platinum Group Elements (PGEs) are contaminants of emerging environmental concern considering their continuous increasing use and subsequent release in the environment. While recent field studies provided PGE levels in seawater, scarce knowledge still exists regarding PGE contamination in marine organisms, especially for rhodium (Rh). Water, macroalgae and mussels were sampled along two representative urbanized estuarine systems and adjacent coastal areas (Douro and Ave estuaries, Portugal). Rhodium and platinum (Pt) concentrations were quantified through both stripping voltammetry and mass spectrometry in collected samples. Spatial mapping of PGE contamination was, to a certain extent, correlated with proxies of urban effluents. The use of Pt/Rh ratios reflected the dominant influence of PGE traffic emissions along the Douro and inputs from various sources (including industries) on the Ave Estuary. Macroalgae and mussels PGE concentrations reflected urban pressure, amplifying environmental signals, and supporting their relevant use as bioindicators of PGE contamination in estuarine/coastal systems., 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.)

Although most deep-sea areas are remote in comparison to coastal zones, a growing body of literature indicates that many sensitive ecosystems could be under increased stress from anthropogenic sources. Among the multiple potential stressors, microplastics (MPs), pharmaceuticals and personal care products (PPCPs/PCPs) and the imminent start of commercial deep-sea mining have received increased attention. Here we review recent literature on these emerging stressors in deep-sea environments and discuss cumulative effects with climate change associated variables. Importantly, MPs and PPCPs have been detected in deep-sea waters, organisms and sediments, in some locations in comparable levels to coastal areas. The Atlantic Ocean and the Mediterranean Sea are the most studied areas and where higher levels of MPs and PPCPs have been detected. The paucity of data for most other deep-sea ecosystems indicates that many more locations are likely to be contaminated by these emerging stressors, but the absence of studies hampers a better assessment of the potential risk. The main knowledge gaps in the field are identified and discussed, and future research priorities are highlighted to improve hazard and 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 Elsevier B.V. All rights reserved.)

Industrial deep-sea mining will release plumes containing metals that may disperse over long distances; however, there is no general understanding of metal effects on marine ecosystems. Thus, we conducted a systematic review in search of models of metal effects on aquatic biota with the future perspective to support Environmental Risk Assessment (ERA) of deep-sea mining. According to results, the use of models to study metal effects is strongly biased towards freshwater species (83% freshwater versus 14% marine); Cu, Hg, Al, Ni, Pb, Cd and Zn are the best-studied metals, and most studies target few species rather than entire food webs. We argue that these limitations restrain ERA on marine ecosystems. To overcome this gap of knowledge, we suggest future research directions and propose a modelling framework to predict the effects of metals on marine food webs, which in our view is relevant for ERA of deep-sea mining., 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.)

Contaminants of emerging concern have been increasingly associated with the modulation of the epigenome, leading to potentially inherited and persistent impacts on apical endpoints. Here, we address the performance of the OECD Test No. 236 FET (fish embryo acute toxicity) in the identification of chemicals able to modulate the epigenome. Using zebrafish (Danio rerio) embryos, acute and chronic exposures were performed with the pharmaceutical, simvastatin (SIM), a widely prescribed hypocholesterolemic drug reported to induce inter and transgenerational effects. In the present study, the epigenetic effects of environmentally relevant concentrations of SIM (from 8 ng/L to 2000 ng/L) were addressed following (1) an acute embryo assay based on OECD Test No. 236 FET, (2) a chronic partial life-cycle exposure using adult zebrafish (90 days), and (3) F1 embryos obtained from parental exposed animals. Simvastatin induced significant effects in gene expression of key epigenetic biomarkers (DNA methylation and histone acetylation/deacetylation) in the gonads of exposed adult zebrafish and in 80 hpf zebrafish embryos (acute and chronic parental intergenerational exposure), albeit with distinct effect profiles between biological samples. In the chronic exposure, SIM impacted particularly DNA methyltransferase genes in males and female gonads, whereas in F1 embryos SIM affected mostly genes associated with histone acetylation/deacetylation. In the embryo acute direct exposure, SIM modulated the expression of both genes involved in DNA methylation and histone deacetylase. These findings further support the use of epigenetic biomarkers in zebrafish embryos in a high throughput approach to identify and prioritize epigenome-modulating chemicals., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The antidiabetic drug Metformin (MET), one of the most prevalent pharmaceuticals in the environment, is currently detected in surface waters in the range of ng/L to low μg/L. As current knowledge regarding the long-term effects of environmentally relevant concentrations of MET in nontarget organisms is limited, the present study aimed at investigating the generational effects of MET, in concentrations ranging from 390 to 14 423 ng/L in the model organism Danio rerio (up to 9 mpf), including the effects on its nonexposed offspring (until 60 dpf). We integrate several apical end points, i.e., embryonic development, survival, growth, and reproduction, with qRT-PCR and RNA-seq analyses to provide additional insights into the mode of action of MET. Reproductive-related parameters in the first generation were particularly sensitive to MET. MET parental exposure impacted critical molecular processes involved in the metabolism of zebrafish males, which in turn affected steroid hormone biosynthesis and upregulated male vtg1 expression by 99.78- to 155.47-fold at 390 and 14 432 MET treatment, respectively, pointing to an estrogenic effect. These findings can potentially explain the significant decrease in the fertilization rate and the increase of unactivated eggs. Nonexposed offspring was also affected by parental MET exposure, impacting its survival and growth. Altogether, these results suggest that MET, at environmentally relevant concentrations, severely affects several biological processes in zebrafish, supporting the urgent need to revise the proposed Predicted No-Effect Concentration (PNEC) and the Environmental Quality Standard (EQS) for MET.

Metformin (MET), an anti-diabetic pharmaceutical of large-scale consumption, is increasingly detected in surface waters. However, current knowledge on the long-term effects of MET on non-target organisms is limited. The present study aimed to investigate the effects of MET in the model freshwater teleost Danio rerio, following a full life-cycle exposure to environmentally relevant concentrations (390 to 14 423 ng/L). Considering that the mode of action (MoA) of MET on non-target organisms remains underexplored and that MET may act through similar human pathways, i.e., lipid and energy metabolisms, biochemical markers were used to determine cholesterol and triglycerides levels, as well as mitochondrial complex I activity in zebrafish liver. Also, the hepatosomatic index as an indication of metabolic disruption, and the expression levels of genes involved in MET's putative MoA, i.e. acaca, acadm, cox5aa, idh3a, hmgcra, prkaa1, were determined, the last by qRT-PCR. A screening of mRNA transcripts, associated with lipid and energy metabolisms, and other signaling pathways potentially involved in MET-induced toxicity were also assessed using an exploratory RNA-seq analysis. The findings here reported indicate that MET significantly disrupted critical biochemical and molecular processes involved in zebrafish metabolism, such as cholesterol and fatty acid biosynthesis, mitochondrial electron transport chain and tricarboxylic acid cycle, concomitantly to changes on the hepatosomatic index. Likewise, MET impacted other relevant pathways mainly associated with cell cycle, DNA repair and steroid hormone biosynthesis, here reported for the first time in a non-target aquatic organism. Non-monotonic dose response curves were frequently detected in biochemical and qRT-PCR data, with higher effects observed at 390 and 2 929 ng/L MET treatments. Collectively, the results suggest that environmentally relevant concentrations of MET severely disrupt D. rerio metabolism and other important biological processes, supporting the need to revise the proposed environmental quality standard (EQS) and predicted no-effect concentration (PNEC) for MET., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Teresa Neuparth reports financial support was provided by FCT - Fundação para a Ciência e a Tecnologia and INTERREG. Miguel Santos reports financial support was provided by FCT - Fundação para a Ciência e a Tecnologia. Susana Barros reports financial support was provided by FCT - Fundação para a Ciência e a Tecnologia. Marlene Pinheiro reports financial support was provided by FCT - Fundação para a Ciência e a Tecnologia. Hugo Morais reports financial support was provided by FCT - Fundação para a Ciência e a Tecnologia. Nelson Alves reports financial support was provided by FCT - Fundação para a Ciência e a Tecnologia. Ana M. Coimbra reports financial support was provided by FCT - Fundação para a Ciência e a Tecnologia. Jose Benito Quintana reports was provided by Xunta de Galicia. Jose Benito Quintana reports financial support was provided by Agencia Estatal de Investigación., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Contaminants of emerging concern (CECs) are compounds of diverse origins that have not been deeply studied in the past which are now accruing growing environmental interest. The NOR-Water project aimed to identify the main CECs and their sources in the water environment of Northern Portugal-Galicia (located in northwest Spain) transnational region. To achieve these goals, a suspect screening analytical methodology based on the use of liquid chromatography coupled to high resolution mass spectrometry (LC-HRMS) was applied to 29 sampling sites in two campaigns. These sampling sites included river and sea water, as well as treated wastewater. The screening was driven by a library of over 3500 compounds, which included 604 compounds prioritized from different relevant lists on the basis of the persistency, mobility, and toxicity criteria. Thus, a total of 343 chemicals could be tentatively identified in the analyzed samples. This list of 343 identified chemicals was submitted to the classification workflow used for prioritization and resulted in 153 chemicals tentatively classified as persistent, mobile, and toxic (PMT) and 23 as very persistent and very mobile (vMvP), pinpointing the relevance of these types of chemicals in the aqueous environment. Pharmaceuticals, such as the antidepressant venlafaxine or the antipsychotic sulpiride, and industrial chemicals, especially high production volume chemicals (HPVC) such as ε-caprolactam, were the groups of compounds that were detected at the highest frequencies.

Due to non-linear interactions, the effects of contaminant mixtures on aquatic ecosystems are difficult to assess, especially under temperature rise that will likely exacerbate the complexity of the responses. Yet, under the current climatic crisis, assessing the effects of water contaminants and temperature is paramount to understanding the biological impacts of mixtures of stressors on aquatic ecosystems. Here, we use an ecosystem model followed by global sensitivity analysis (GSA) to prioritize the effects of four single emerging contaminants (ECs) and their mixture, combined with two temperature rise scenarios, on the biomass production of a NE Atlantic estuary. Scenarios ran for 10 years with a time-step of 0.1 days. The results indicate that macroinvertebrate biomass was significantly explained by the effect of each single EC and by their mixture but not by temperature. Globally, the most adverse effects were induced by two ECs and by the mixture of the four ECs, although the sensitivity of macroinvertebrates to the tested scenarios differed. Overall, the present approach is useful to prioritize the effects of stressors and assess the sensitivity of the different trophic groups within food webs, which may be of relevance to support decision making linked to the sustainable management of estuaries and other aquatic systems.

Hazard assessment strategies are often supported by extrapolation of damage probabilities, regarding chemical action and species susceptibilities. Yet, growing evidence suggests that an adequate sampling of physiological responses across a representative taxonomic scope is of paramount importance. This is particularly relevant for Nuclear Receptors (NR), a family of transcription factors, often triggered by ligands and thus, commonly exploited by environmental chemicals. Within NRs, the ligand-induced Ecdysone Receptor (EcR) provides a remarkable example. Long regarded as arthropod specific, this receptor has been extensively targeted by pesticides, seemingly innocuous to non-target organisms. Yet, current evidence clearly suggests a wider presence of EcR orthologues across metazoan lineages, with unknown physiological consequences. Here, we address the state-of-the-art regarding the phylogenetic distribution and functional characterization of metazoan EcRs and provide a critical analysis of the potential disruption of such EcRs by environmental chemical exposure. Using EcR as a case study, hazard assessment strategies are also discussed in view of the development of a novel "precision hazard assessment paradigm.

The objective of the present study was to evaluate the potential toxic effects of polyethylene microplastics (PE-MPs) (40-48 μm) on the Mediterranean mussel Mytilus galloprovincialis in controlled laboratory conditions. The exposure was carried out for 14 days with three environmentally relevant PE-MPs concentrations of 1, 10, and 100 and a high concentration of 1000 μg/L. Effects of PE-MPs were assessed by evaluating the filtration rate (FR) after 7 and 14 days of exposure and by analyzing biochemical biomarkers of oxidative stress (catalase - CAT, glutathione S-transferase - GST, and the levels of lipid peroxidation - LPO) in the M. galloprovincialis digestive gland after 14 days of exposure. Results showed that M. galloprovincialis does not accumulate PE-MPs of 40-48 μm size in its whole tissues. The filtration rate was significantly reduced with the increase of PE-MPs concentrations. The biochemical biomarkers indicated that PE-MPs induced oxidative damage (LPO) at low concentrations (1 and 10 μg/L) with a significant reduction in females of 1000 μg/L treated group and inactivate antioxidative system (CAT and GST) in the digestive gland of both sexes at high concentrations (100 and 1000 μg/L). This study demonstrates that PE-MPs have biological effects on M. galloprovincialis at environmentally relevant concentrations thus brings new insights on the potential impacts of PE-MPs in marine bivalves.

The deep-sea is the biggest ecosystem in the world and despite the extreme conditions that characterize it, is highly biodiverse and complex. Deep-sea mining has been foreseen as a potential and concerning new stressor, and among the deep-sea mining associated stressors, sediment plumes, likely to be released into the water column as a side effect of mining, can reach habitats within a radius of more than a hundred kilometers. The present study examined the effects of suspended sediments of different grain sizes (63-125 μm, 125-250 μm and 250-500 μm) in the model species Mytilus galloprovincialis, at 4 bar, as a proxy to address the potential effects of sediment plumes, in the water column, with different grain sizes under high pressure conditions. Functional (filtration rate - FR), biochemical (catalase - CAT, glutathione s-transferase - GST, lipid peroxidation - LPO) and molecular (gene expression of [actin (ACTN), glutathione S-transferase alpha (GSTA), superoxide dismutase 2 (SOD2), catalase (CAT), heat shock protein 60 (HSP60), cytochrome c oxidase (COI) and DNA mismatch repair protein (MSH6)]) endpoints were studied in juvenile organisms. The FR decreased significantly for all tested grain size ranges, with a more severe effect for the particles with a diameter between 63 and 125 μm. In addition to the FR, significant changes were also observed for all tested biomarkers. Gene expression was significantly downregulated for CAT and ACTN. Overall, this study demonstrated that the smaller sized particles are the ones leading to more severe effects. Given their high dispersion potential and longer suspension periods under mining operation scenarios, particular attention should be given to the release of sediment plumes that may affect deep-sea environments and the water column. It is, therefore, vital to create standards and guidelines for sustainable mining practices., (Copyright © 2021 Elsevier Inc. All rights reserved.)

The adverse effects of certain environmental chemicals have been recently associated with the modulation of the epigenome. Although changes in the epigenetic signature have yet to be integrated into hazard and risk assessment, they are interesting candidates to link environmental exposures and altered phenotypes, since these changes may be passed across multiple non-exposed generations. Here, we addressed the effects of simvastatin (SIM), one of the most prescribed pharmaceuticals in the world, on epigenetic regulation using the amphipod Gammarus locusta as a proxy, to support its integration into hazard and environmental risk assessment. SIM is a known modulator of the epigenome in mammalian cell lines and has been reported to impact G. locusta ecological endpoints at environmentally relevant levels. G. locusta juveniles were exposed to three SIM environmentally relevant concentrations (0.32, 1.6 and 8 µg L -1 ) for 15 days. Gene transcription levels of selected epigenetic regulators, i.e., dnmt1, dmap1, usp7, kat5 and uhrf1 were assessed, along with the quantification of DNA methylation levels and evaluation of key ecological endpoints: survival and growth. Exposure to 0.32 and 8 µg L -1 SIM induced significant downregulation of DNA methyltransferase 1 (dnmt1), concomitant with global DNA hypomethylation and growth impacts. Overall, this work is the first to validate the basal expression of key epigenetic regulators in a keystone marine crustacean, supporting the integration of epigenetic biomarkers into hazard assessment frameworks., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

The use of transcriptomics data brings new insights and works as a powerful tool to explore the molecular mode of action (MoA) of transgenerational inheritance effects of contaminants of emerging concern. Therefore, in this dataset, we present the transcriptomic data of the transgenerational effects of environmentally relevant simvastatin levels, one of the most prescribed human pharmaceuticals, in the keystone amphipod species Gammarus locusta . In summary, G. locusta juveniles were maintained under simvastatin exposure up to adulthood (exposed group - F0E) and the offspring of F0E were transferred to control water for the three subsequent generations (transgenerational group - F1T, F2T and F3T). To gain insights into the biological functions and canonical pathways transgenerationally disrupted by simvastatin, a G. locusta de novo transcriptome assembly was produced and the transcriptomic profiles of three individual G. locusta females, per group, over the four generations (F0 to F3) - solvent control groups (F0.C, F1.C, F2.C and F3.C), F0 320 ng/L simvastatin exposed group (F0.320E) and F1 to F3 320 transgenerational group (F1.320T; F2.320T and F3.320T) - were analyzed. Briefly, Illumina HiSeq™ 2500 platform was used to perform RNA sequencing, and due to the unavailability of G. locusta genome, the RNA-seq datasets were assembled de novo using Trinity and annotated with Trinotate software. After assembly and post-processing steps, 106093 transcripts with N50 of 2371 bp and mean sequence length of 1343.98 bp was produced. BUSCO analyses showed a transcriptome with gene completeness of 97.5 % Arthropoda library profile. The Bowtie2, RSEM and edgeR tools were used for the differential gene expression (DEGs) analyses that allowed the identification of a high quantity of genes differentially expressed in all generations. Finally, to identify the main metabolic pathways affected by the transgenerational effects of SIM across all generations, the DGEs genes were blasted onto KEGG pathways database using the KAAS webserver. The data furnished in this article allows a better molecular understanding of the transgenerational effects produced by simvastatin in the keystone amphipod G. locusta and has major implications for hazard and risk assessment of pharmaceuticals and other emerging contaminants. This article is related to the research article entitled "Transgenerational inheritance of chemical-induced signature: a case study with simvastatin [1]., Competing Interests: 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., (© 2020 The Author(s).)

Simvastatin (SIM), a hypocholesterolaemic drug belonging to the statins group, is a widely prescribed pharmaceutical for prevention of cardiovascular diseases. Several studies showed that lipophilic statins, as SIM, cross the blood-brain barrier and interfere with the energy metabolism of the central nervous system in humans and mammalian models. In fish and other aquatic organisms, the effects of SIM on the brain energy metabolism are unknown, particularly following exposure to low environmentally relevant concentrations. Therefore, the present study aimed at investigating the influence of SIM on gene signaling pathways involved in brain energy metabolism of adult zebrafish ( Danio rerio ) following chronic exposure (90 days) to environmentally relevant SIM concentrations ranging from 8 ng/L to 1000 ng/L. Real-time PCR was used to determine the transcript levels of several genes involved in different pathways of the brain energy metabolism ( glut1b, gapdh, acadm, accα, fasn, idh3a, cox4i1 , and cox5aa ). The findings here reported integrated well with ecological and biochemical responses obtained in a parallel study. Data demonstrated that SIM modulates transcription of key genes involved in the mitochondrial electron transport chain, in glucose transport and metabolism, in fatty acid synthesis and β-oxidation. Further, SIM exposure led to a sex-dependent transcription profile for some of the studied genes. Overall, the present study demonstrated, for the first time, that SIM modulates gene regulation of key pathways involved in the energy metabolism in fish brain at environmentally relevant concentrations.

The increasing use of Sertraline (SER) as antidepressant and its consequent presence in the aquatic environment is raising concern about the chronic effects of this pharmaceutical to aquatic organisms. As the current concentrations of SER in surface waters are typically in the low ng/L range, acute toxicity is unlikely to occur. However, prolonged exposure to low concentrations of SER may lead to sub-lethal effects in aquatic organisms, including alterations in important physiological functions like growth, reproduction, behaviour, and also in key biochemical processes, such as those associated with neurotransmission and redox balance. To test this hypothesis, we selected the amphipod Gammarus locusta, a keystone species used in ecotoxicological hazard assessment. In the present study, juveniles' G. locusta from a permanent laboratory culture were chronically exposed to low concentrations of SER (8-1000 ng/L) in a bioassay that lasted for 48 days, allowing for a life-cycle study including effects on reproduction. At the lowest SER concentrations with environmental relevance (8, 40 and 200 ng/L) we detected no significant changes in key ecological endpoints such as survival, growth, reproduction and movement behaviour, or in any of the biochemical markers analysed. However, at 1000 ng/L SER (a concentration one order of magnitude higher than the levels reported in aquatic environments) females showed a significant increase in movement versus control, whereas no activity changes were observed in males. Overall, these findings indicate that G. locusta females are potentially more susceptible to the chronic effects of SER. Moreover, the current environmental SER concentrations are unlikely to affect amphipod's ecological endpoints because only SER concentrations higher than the levels reported in aquatic environments produced effects on the behaviour of G. locusta females. However, the increasing consumption of SER, highlights the importance of monitoring its chronic risk to the aquatic wildlife., (Copyright © 2019 Elsevier Inc. All rights reserved.)

The deep-sea is the biggest ecosystem in the world and is characterized by extreme conditions such as high pressure, low temperatures and absence or limited light. Despite the scarce studies due to inaccessibility, these ecosystems are considered highly biodiverse. The deep-sea is subjected to anthropogenic stressors with deep-sea mining being a likely new form of disruption. Understanding how it affects the surrounding environments is paramount to develop guidelines to protect sensitive habitats and allow for responsible exploitation of resources. One of the potential stressors associated with deep-sea mining are the sediment laden plumes that can be generated during the mining process. The present study examined, for the first time, the effects of suspended sediments (0, 1, 2 and 4 g/L) in the model mussel species, Mytilus galloprovincialis, under hyperbaric conditions (1, 4 and 50 Bar). Functional endpoints, i.e. feeding assays, together with biochemical biomarkers of oxidative stress [catalase (CAT), lipid peroxidation (LPO), glutathione-s-transferase (GST) and superoxide dismutase (SOD)] were studied in juvenile mussels. The filtration rate (FR) of M. galloprovincialis decreased with the increment in the sediment concentrations, for all tested pressure conditions (1, 4 and 50 Bar). Significant alterations were also observed for all tested biomarkers, being sediment and pressure-dependent. Interestingly, pressure had an effect in GST activity, that increased in the 4 and 50 Bar experiments in comparison with the results at 1 Bar. Remarkably, filtration rates were significantly affected by pressure. These findings will support the filling of the knowledge gaps related with the hazard assessment of deep-sea mining associated stressors., (Copyright © 2019 Elsevier B.V. All rights reserved.)

An innovative photoreactor, FluHelik, was used to promote the degradation of contaminants of emerging concern (CECs) by a photochemical UVC/H 2 O 2 process. First, the system was optimized for the oxidation of a model antibiotic, oxytetracycline (OTC), using both ultrapure water (UPW) and a real urban wastewater (UWW) (collected after secondary treatment) as solution matrices. Following, the process was evaluated for the treatment of a UWW spiked with a mixture of OTC and 10 different pharmaceuticals established by the Swiss legislation at residual concentrations (∑CECs <660 μg L -1 ). The performance of the FluHelik reactor was analyzed both at lab and pre-pilot scale in multiple and single pass flow modes. The efficiency of the FluHelik photoreactor, at lab-scale, was evaluated at different operational conditions (H 2 O 2 concentration, UVC lamp power (4, 6 and 11 W) and flow rate) and further compared with a conventional Jets photoreactor. Both photoreactors exhibited similar OTC removal efficiencies at the best conditions; however, the FluHelik reactor showed to be more efficient (1.3 times) in terms of mineralization when compared with the Jets reactor. Additionally, the efficiency of the UVC/H 2 O 2 photochemical system using the FluHelik photoreactor in reducing the toxicity of the real effluent containing 11 pharmaceuticals was evaluated through zebrafish (Danio rerio) embryo toxicity bioassays. FluHelik scale-up from laboratory to pre-pilot to promote UVC/H 2 O 2 photochemical process proved to be feasible., (Copyright © 2019. Published by Elsevier B.V.)

We aimed to show how the predicted pH decrease in the ocean would alter the toxicity, bioconcentration and dietary transfer of trace metal copper on seagrass ecosystems, on a short-term basis. Seagrass Zostera noltei was exposed to two pH levels (8.36 and 8.03) and three copper levels (nominal concentrations, <3, 30 and 300 μg Cu L -1 ) in a factorial design during 21 days, while Gammarus locusta amphipods were continuously fed with the treated seagrass leaves. We found that the toxicity and bioconcentration of copper in seagrasses were not affected by pH, yet complex copper-pH interactions were observed in the seagrass photosynthesis. We demostrated that seagrasses can act as a copper source in the food web via direct consumption by herbivores. Future research need to investigate the interactive effects on a long-term basis, and to include biochemical and molecular endpoints to provide additional insights to the complex phisiological interactions observed., (Copyright © 2019. Published by Elsevier Ltd.)

This work deals with a preliminary study aimed at evaluating the efficiency of three different microalgae strains, namely Chlorella sorokiniana, Chlorella vulgaris and Scenedesmus obliquus in the bioremediation of diclofenac contaminated water. For this purpose, microalgae were cultured in bubbling column photobioreactors (PBRs) under batch operation until the end of the exponential growth phase. For the three strains, the concentration of diclofenac in the PBRs aquatic medium decreased along microalgae growing, which pointed to biodegradation as the main removal mechanism. Among the three strains, S. obliquus was the most capable to reduce diclofenac concentration (99% removal from an initial concentration of 25,000 μg l -1 ). However, such a large removal does not guarantee an efficient treatment since transformation products (TPs) exceeding the concentration and/or toxicity of the parent compound may be generated during biodegradation of diclofenac. Thus, for a comprehensive evaluation of the microalgae treatments efficiency, the final effluents from the PBRs were tested for their effects on the embryonic development of zebrafish. Again, the S. obliquus treatment was the most efficient in the reduction of toxicity, with the corresponding effluents having no effects on the embryo's mortality or abnormalities incidence (at 80 h post fertilization). In any case, for the three strains, the toxicity effects of effluents were equal or lower than those determined for diclofenac solutions with the same concentration. Therefore, it may be stated that, at the end of the batch culture, the removal of diclofenac by the considered strains did not involve the generation of toxic TPs to zebrafish embryos., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Simvastatin (SIM), a hypocholesterolaemic compound, is among the most prescribed pharmaceuticals for cardiovascular disease prevention worldwide. Several studies have shown that acute exposure to SIM causes multiple adverse effects in aquatic organisms. However, uncertainties still remain regarding the chronic effects of SIM in aquatic ecosystems. Therefore, the present study aimed to investigate the effects of SIM in the model freshwater teleost zebrafish (Danio rerio) following a chronic exposure (90 days) to environmentally relevant concentrations ranging from 8 ng/L to 1000 ng/L. This study used a multi-parameter approach integrating distinct ecologically-relevant endpoints, i.e. survival, growth, reproduction and embryonic development, with biochemical markers (cholesterol and triglycerides). Real Time PCR was used to analyse the transcription levels of key genes involved in the mevalonate pathway (hmgcra, cyp51, and dhcr7). Globally, SIM induced several effects that did not follow a dose-response relationship; embryonic development, biochemical and molecular markers, were significantly impacted in the lower concentrations, 8 ng/L, 40 ng/L and/or 200 ng/L, whereas no effects were recorded for the highest tested SIM levels (1000 ng/L). Taken together, these findings expand our understanding of statin effects in teleosts, demonstrating significant impacts at environmentally relevant concentrations and highlight the importance of addressing the effects of chemicals under chronic low-level concentrations., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Given the lack of knowledge regarding climate change-chemical exposure interactions, it is vital to evaluate how these two drivers jointly impact aquatic species. Thus, for the first time, we aimed at investigating the combined effects of increased temperature, pCO 2 and the synthetic progestin levonorgestrel on survival, growth, consumption rate and reproduction of the amphipod Gammarus locusta. For that, a full factorial design manipulating temperature [ambient temperature and warming (+4 °C)], pCO 2 [normocapnia and hypercapnia (Δ pH 0.5 units)] and the progestin levonorgestrel (LNG: L1 - 10 ngLL -1 and L2 - 1000 ngLL -1 , control - no progestin and solvent control - vehicle ethanol (0.01%)) was implemented for 21 days. G. locusta was strongly negatively affected by warming, experiencing higher mortality rates (50-80%) than in any other treatments. Instead, growth rates were significantly affected by interactions of LNG with temperature and pCO 2 . It was observed, in the short-term (7d) that under ambient temperature (18 °C) and hypercapnic conditions (pH 7.6), the LNG presence promoted the amphipod's growth, while in the medium-term (21d) this response was not observed. Relative consumption rates (RCRs), during the first week were higher than in the third week. Furthermore, in the first week, RCRs were negatively affected by higher temperature while in the third week, RCRs were negatively affected by acidification. Furthermore, it was observed a negative effect of higher temperature and acidification on G. locusta fecundity, contrarily to LNG. Concluding, the impact of increased temperature and pCO 2 was clearly more adverse for the species than exposure to the synthetic progestin, however, some interactions between the progestin and the climate factors were observed. Thus, in a future scenario of global change, the presence of LNG (and other progestins alike) may modulate to a certain level the effects of climate drivers (and vice-versa) on the gammarids fitness and reproduction., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Methyl farnesoate (MF) is an arthropod hormone that plays a key role in the physiology of several arthropods' classes being implicated in biological processes such as molting and reproduction. The development of an analytical technique to quantify the levels of this compound in biological tissues can be of major importance for the field of aquaculture/apiculture conservation and in endocrine disruption studies. Therefore, the aim of this study was to develop a simple and sensitive method to measure native levels of MF in the tissue of three representative species from different arthropods classes with environmental and/or economic importance. Thus, a new approach using whole organisms and the combination of matrix solid-phase dispersion with gas chromatography coupled to mass spectrometry was developed. This method allows quantifying endogenous MF at low levels (LOQs in the 1.2-3.1ng/g range) in three arthropod species, and could be expanded to additional arthropod classes. The found levels ranged between 2 and 12ng/g depending on the studied species and gender. The overall recovery of the method was evaluated and ranged between 69 and 96%., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Triclocarban (TCC), a common antimicrobial agent widely used in many household and personal care products, has been widely detected in aquatic ecosystems worldwide. Due to its high lipophilicity and persistence in the aquatic ecosystems, TCC is of emerging environmental concern. Despite the frequently reported detection of TCC in the environment and significant uncertainties about its long term effects on aquatic ecosystems, few studies have addressed the chronic effects of TCC in aquatic organisms at ecologically relevant concentrations. Therefore, we aimed at testing a broad range of biological responses in the amphipod Gammarus locusta following a chronic (60 days) exposure to environmentally relevant concentrations of TCC (100, 500 and 2500ng/L). This work integrated biochemical markers of oxidative stress (catalase (CAT), glutathione-s-transferase (GST) and lipid peroxidation (LPO)) and neurotransmission (acetylcholinesterase (AChE)) with several key ecological endpoints, i.e. behaviour, survival, individual growth and reproduction. Significant alterations were observed in all biochemical markers. While AChE showed a dose-response curve (with a significant increased activity at a TCC concentration of 2500ng/L), oxidative stress markers did not follow a dose-response curve, with significant increase at 100 and/or 500ng/L and a decreased activity in the highest concentration (2500ng/L). The same effect was observed in the females' behavioural response, whereas males' behaviour was not affected by TCC exposure. The present study represents a first approach to characterize the hazard of TCC to crustaceans., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Literature data and data obtained with modelling tools were compiled to derive the physicochemical behaviour of 24 priority Hazardous and Noxious Substances (HNS), as a proxy to improve environmental, public health and political issues in relation to HNS spills. Parameters that rule the HNS behaviour in water and those that determine their distribution and persistence in the environment, such as fugacity, physicochemical degradation, biodegradation, bioaccumulation/biotransformation and aquatic toxicity, were selected. Data systematized and produced in the frame of the Arcopol Platform project was made available through a public database (http://www.ciimar.up.pt/hns/substances.php). This tool is expected to assist stakeholders involved in HNS spills preparedness and response, policy makers and legislators, as well as to contribute to a current picture of the scientific knowledge on the fate, behaviour, weathering and toxicity of priority HNS, being essential to support future improvements in maritime safety and coastal pollution response before, during and after spill incidents., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Emerging pollutants, such as pharmaceuticals, may pose a considerable environment risk. Hypocholesterolaemic drugs such as statins are among the most prescribed human pharmaceuticals in western European countries. In vertebrates, this therapeutic class disrupts the cholesterol synthesis by inhibiting the enzyme 3-hydroxy-3-methyl-glutaryl-CoA reductase (HMGR), responsible for the limiting step in the mevalonate pathway. Recently, functional studies have shown that statins competitively inhibit HMGR in vertebrates and arthropods, two taxa that have diverged over 450 million years ago. Importantly, chronic simvastatin exposure disrupts crustacean reproduction and development at environmentally relevant concentrations. Hence, a fundamental question emerges: what is the taxonomic scope of statins-induced HMGR inhibition across metazoans? Here, we address this central question in a large sampling of metazoans using comparative genomics, homology modelling and molecular docking. Sequence alignment of metazoan HMGRs allowed the annotation of highly conserved catalytic, co-factor and substrate binding sites, including residues highjacked for statin binding. Furthermore, molecular docking shows that the catalytic domains of metazoan HMGRs are highly conserved regarding interactions, not only with HMG-CoA, but also with both simvastatin and atorvastatin, the top prescribed statins in Europe and USA. Hence, the data indicates that both statins are expected to competitively inhibit metazoan's HMGRs, and therefore all metazoan taxa might be at risk. The environmental relevance of these findings are discussed and research priorities established. We believe that the conceptual framework used in this study can be applied to other emerging pollutants and assist in the design of toxicity testing and risk assessment., (Copyright © 2016 Elsevier B.V. All rights reserved.)