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The effects of metallic nanoparticles (NPs) on the estuarine biota have mostly been shown for concentrations higher than those actually measured or predicted in these environments. To address this gap, a range of concentrations expected to occur in estuarine environments (from 0.01 to 1 μg g−1) was employed in microcosms studies to assess the impact of Cu NPs in the denitrification pathway. This was achieved by quantifying the gene expression in estuarine sediments exposed to Cu NPs for up to six days and potential denitrification rates. The expression of nitrite (nirS) and nitrous oxide (nosZ) reductase genes was enhanced in a timewise manner upon exposure to Cu NPs

This work aimed to investigate the possible effect of 2 quenchers commonly used in H2O2-based advanced oxidation technologies (AOTs), i.e. catalase and sodium sulphite (Na2SO3), on the analytical signal of 3 detectors coupled to liquid chromatography (LC): tandem mass spectrometry (LC-MS/MS), fluorescence detection (LC-FD) and LC-diode array detection (LC-DAD). The observation of analytical interferences for a group of compounds when studying the removal by continuous mode UV/H2O2 of 26 micropollutants (MPs) from a spiked surface water (SW), for which the residual H2O2 in the samples was quenched by Na2SO3, triggered the need of understanding these effects and thus catalase was used as comparative quencher. From the 26 MPs having a wide range of polarity and pKa, those monitored after electrospray ionization (ESI) under positive ionization (PI) mode and presenting a pKa higher than 5.9 revealed a great signal suppression, but only when using Na2SO3 as H2O2 quencher. In this sense, we further explored this effect by selecting 2 MPs, metoprolol and diclofenac, which had respectively signal suppression and no interference in the LC-MS/MS response. These MPs were analysed before and after addition of H2O2 and catalase or Na2SO3 in reaction vials, using: (i) different detectors coupled to LC, namely LC-MS/MS with ESI under PI and negative ionization (NI) modes, LC-FD and LC-DAD; (ii) different environmental matrices (SW, drinking water, wastewater) and ultrapure water; and (iii) different magnitude levels (0.1–10 mg L−1). The results demonstrated a remarkable signal suppression in LC-MS/MS analyses under PI mode for those compounds with pKa higher than 5.9, confirming the interfering effect of H2O2/Na2SO3. To the best of our knowledge, the analytical interference in the LC-MS/MS analysis, after adding Na2SO3 to quench H2O2 in AOTs experiments was never reported before and the results presented herein support the recommendation to use catalase instead of Na2SO3 as quencher in AOTs studies.

The staff of the XAFS beamline BM30B (FAME) at ESRF, France, and more particularly Dr. D. Testemale, are gratefully acknowledged for their help in performing the XAS experiments.; International audience; Application of anaerobically digested sludges and manure (digestates) to agricultural fields reduces the need for fertilizer application, but might also pose environmental risks due to the introduction of various pollutants –including metal trace elements and pharmaceuticals– to amended soils. The simultaneous presence of different contaminants can affect plant uptake, altering the performance of phytoremediation. This work aims to investigate and compare the single and combined effects of Enrofloxacin (Enro) and Cu on their plant uptake from digestate solutions. Plant uptake experiments were carried out by exposing, for 5 days, Phragmites australis to three different concentrations of a liquid digestate doped with Cu and/or Enro. Contaminant concentrations were afterward determined in both plant tissues and digestate solutions. Contaminant speciation in solution and Cu speciation in plant roots were also assessed. Cu plant uptake (mostly in plant roots, Cu concentration varied between 55 and 254 μg·g−1) was observed, especially at low concentration of digestate. At high digestate concentration, Cu uptake decreased due to the formation of Cu-DOC complexes rendering Cu less bioavailable. The presence of the antibiotic slightly further reduced Cu plant uptake due to the formation of Cu-Enro complexes. Plant roots also accumulated Enro, an accumulation also higher for the lowest concentration of digestate (an increase of 40%), and that slightly increased in the presence of Cu (up to 226 μg·g−1). However, proportion of free Enro in solution decreased (up to 70%) in the presence of Cu due to complexation. Cu speciation in plant roots confirmed the implication of the complexation in the uptake of Cu and Enro when they are simultaneously present in solution. Thus, the presence of amendments (such as digestates) increasing the DOC content, in addition to the interactions between contaminants, should be taken into account for field crop soils as well as for phytoremediation technologies.

This study aimed to evaluate the potential of CWs to remove bacteria, including antibiotic resistant bacteria, and antibiotics from livestock (swine production) wastewater. Microcosms were assembled with Phragmites australis (Cav.) Trin. ex Steud and feed with wastewater spiked or not with 100 µg/L of enrofloxacin or of ceftiofur, alone or as a mixture. Wastewater (spiked or not) was treated during twenty one-week cycles. Treated wastewater was collected at weeks 1, 2, 4, 8, 14, 18 and 20 to evaluate the removal of pollutants, including nutrients, organic matter, metals and added antibiotics, and total cultivable and antibiotic resistant bacteria (enterobacteria and enterococci), which were enumerated in plate culture media. After treatment, a significant reduction in pollutants load (up to 99% depending on the parameter) was observed regardless of the presence of the veterinary antibiotics. Antibiotics were also significantly removed (removal > 90%). High removal rates were achieved for enterobacteria and enterococci bacteria during the experimental period, in the presence or absence of antibiotics (mean removals between 86 and 100%). For antibiotic resistant bacteria initially present in the influent wastewater mean removals > 90% were obtained, and no significant effect of time or of antibiotic presence was observed. This study is, to our knowledge, the first to focus on the removal of enrofloxacin and ceftiofur resistance in faecal bioindicators in CWs fed with livestock wastewater. Results confirm CWs as a green alternative to reduce the risks associated with the release of livestock effluents into the environment.

Estuaries are productive environments used by many fish as nursery grounds. The initial stages of fishes are highly vulnerable to (a)biotic factors, and anthropogenic pressures, influencing fish larvae assemblages along the estuary. Microplastics (MPs 5 mm) are particularly dangerous to early life stages of fishes because their ingestion can induce gut blockage, limiting food intake or exposing organisms to contamination due to MPs capacity to absorb pollutants. Present work aimed to investigate the contamination of an urban impacted estuary (Douro estuary, NW Portugal) by MPs, and study the abundance and distribution of MPs and fish larvae in this estuary. Monthly sampling surveys were performed from December 2016 to December 2017, in nine stations along the estuary. Sub-surface planktonic horizontal trawls were performed to collect fish larvae and MPs. Planktonic samples were sorted, and fish larvae identified. MPs density was determined using a protocol optimized in our laboratory. A total of 1498 fish larvae belonging to 32 taxa were collected, with a mean density of 11.66 fish larvae 100 m

This study aimed to investigate the potential of microbial communities from the rhizosediment of two plants - Phragmites australis and Juncus maritimus - occurring in an estuarine area subjected to a high anthropogenic impact, to biodegrade ENR, a commonly used veterinary antibiotic. An enrichment process with 1 mgL−1 of ENR was conducted during ca. 9 months, using acetate as a co-substrate. After this, the enriched microbial consortia were challenged with higher ENR concentrations of 2 and 3 mgL−1. Microbial cultures enriched with 1 mgL−1 of ENR were capable of biodegrading this antibiotic, though not completely. By the end of the enrichment phase, microbial cultures were defluorinating an average of 50% of the ENR supplemented. Higher ENR concentrations led to lower biodegradation performances, suggesting a possible toxic/inhibitory effect in the microbial cultures. Phylogenetic identification of the microorganisms isolated from microbial cultures enriched with ENR revealed a high taxonomical diversity, with microorganisms belonging mainly to Proteobacteria and Bacteroidetes phyla. Assemblage of the obtained isolated strains (according to the enriched cultures from which they were isolated) revealed that the resulting consortia were also capable of degrading ENR, indicating that the main microbial players in the biodegradation of this antibiotic were isolated. These consortia also showed to be more robust to degrade higher concentrations of ENR than the corresponding enriched cultures. This study shows that microorganisms derived from rhizosediments of the selected plants, exhibit capacity to biodegrade ENR, though not completely for the concentrations tested, and may be further explored for the development of bioremediation strategies for the treatment of this antibiotic.

This work investigated the potential of microbial communities native to an estuarine environment to biodegrade enrofloxacin (ENR) and oxytetracycline (OXY). Sediments collected from two sites in the Douro river estuary (Porto, Portugal) were used as inocula for the biodegradation experiments. Experiments were carried out for one month, during which ENR and OXY (1 mg L−1) were supplemented individually or in mixture to the cultures at 10-day intervals. Acetate (400 mg L−1) was added to the cultures every 3 days to support microbial growth. A series of experimental controls were established in parallel to determine the influence of abiotic breakdown and adsorption in the removal of the antibiotics. Removal of antibiotics was followed by measuring their concentration in the culture medium. Additionally, next-generation sequencing of the 16S rRNA gene amplicon was employed to understand how microbial communities responded to the presence of the antibiotics. At the end of the biodegradation experiments, microbial cultures derived from the two estuarine sediments were able to remove up to 98% of ENR and over 95% of OXY. The mixture of antibiotics did not affect their removal. ENR was removed mainly by biodegradation, while abiotic mechanisms were found to have a higher influence in the removal of OXY. Both antibiotics adsorbed at different extents to the estuarine sediments used as inocula but exhibited a higher affinity to the sediment with finer texture and higher organic matter content. The presence of ENR and OXY in the culture media influenced the dynamics of the microbial communities, resulting in a lower microbial diversity and richness and in the predominance of bacterial species belonging to the phylum Proteobacteria. Therefore, microbial communities native from estuarine environments have potential to respond to the contamination caused by antibiotics and may be considered for the recovering of impacted environments through bioremediation.

One of the most used protocols to extract and quantify MPs is NOAA protocol in aquatic environments. However, there is still no standardized method to extract and quantify MPs in estuarine waters. The aim of this work was to adapt the NOAA protocol to quantify microplastics in estuarine water and provide all the details and changes to improve the efficiency of the method. For that, four types of plastic (PE-LD; PET; PA; PE-HD) were used in artificial samples to test all the steps of the protocol. Several criteria were tested, namely: (i) quantities of H2O2 used for organic matter degradation; (ii) temperatures of drying samples; and (iii) density separation efficacy. With the proposed modifications, the microplastics extraction were above 90%, regardless the type of plastic, with PE-LD reaching 100% of efficiency. The new adapted protocol that we propose will allow a better efficiency in extraction and quantification of microplastics in samples from estuarine environments. • Four different types of plastic (PE-LD; PET; PA; PE-HD) were used to test the efficiency of the protocol • Details as the ideal quantity of H2O2, temperature and exact quantity of NaCl were tested and defined during the experiments • Efficiency of the microplastics extraction were above 90% Method name: Estuarine waters microplastics quantification protocol, Keywords: Microplastics, Estuarine waters, Contamination, Analytical procedure

Contamination derived from maritime transportation is not only linked to petroleum-based substances but also to transportation of other substances such as hazardous and noxious nature (HNS). Over the last decade, the maritime trade of HNS has been augmenting. However, levels of HNS in aquatic environment are still quite unknown. So, an integrative study, combining a chemical and a multi-biomarker evaluation, was done in the NW Atlantic Iberian coast where specimens of Mytilus sp. were collected. Ten HNS were found in the digestive gland of mussels from Vila Cha and Foz, though at generally low levels. Only measurable levels of benzene, trichloroethylene and 1,1,2-trichlorethane were found in other locations. Principal component analysis produced a clear distinction between Vila Cha and Foz and the remaining sampling sites. Biomarkers associated to contamination were the levels of lipid peroxidation (LPO) and activity of glutathione reductase (GR), superoxide dismutase (SOD) and glutathione peroxidase (GPx) enzymes. Correlations between biomarkers and HNS levels highlight the importance of performing biomonitoring campaigns integrating chemical analysis and a biomarker approach to accurately assess the environmental state of aquatic environments.

The aim of this study focused on the evaluation of constructed wetlands (CWs) microcosms, on a laboratory scale, for the removal of metals from a pig industry effluent while maintaining effluent organic matter and nutrients levels for its later used as a fertilizer. CWs with different macrophytes (Phragmites australis and Typha latifolia) and different substrates (light expanded clay aggregate and lava rock) were tested. Results showed high removals of metals during CWs treatment, with removal rates reaching &gt, 80% for Cd, Cr, Cu, Fe, Mn, and Zn after 2 days of treatment in CWs planted with T. latifolia and &gt, 60% in CWs planted with P. australis. Significant differences were only found between substrates for Fe and Mn in CWs with P. australis. Removal of organic matter (through chemical oxygen demand (COD)) was &gt, 77%, with no significant differences between substrates or plants. Removals of ammonium and phosphate ions ranged between 59&ndash, 84% and 32&ndash, 92%, respectively, in CWs with P. australis and 62&ndash, 75% and 7&ndash, 68% in CWs with T. latifolia, with no significant differences between substrates. Overall, CWs showed potential to be efficient in removing toxic contaminants, as metals, while maintaining moderated levels of nutrients, allowing the use of reclaimed water in agriculture, namely as fertilizer. If one aims for a short CW treatment, CW planted with T. latifolia and expanded clay as substrate could be the more suitable choice.

Biodegradation of two highly persistent fluorinated fungicides, epoxiconazole (EPO) and fludioxonil (FLU), by microbial consortia enriched from estuarine sediment and agricultural soil is reported. After an enrichment period of 6 months, four microbial consortia were able to completely remove and defluorinate the fungicides in co-metabolic conditions. Defluorination was biologically mediated and results suggest it is not a primary catabolic step, as fungicide removal was always faster than its defluorination. Three of the four enriched consortia had similar biodegradation performances in the absence of a co-substrate. Biodegradation kinetics revealed that microbial degradation followed a first-order kinetics, with cultures being capable of biodegrading concentrations up to 10 mg L−1 of EPO or FLU, in a maximum of 21 days. Estimated half-life values for these compounds were significantly lower than those reported in literature, highlighting the unique metabolic performance of the obtained consortia. Analysis of their microbial composition revealed that they integrate several bacterial species belonging to the Proteobacteria phylum, with the most common genera being Pseudomonas, Ochrobactrum and Comamonas. This is the first study providing clear evidence on the biodegradation of EPO and FLU, opening doors for the design of bioremediation technologies for the recovery of ecosystems polluted with such recalcitrant compounds.

Microcystis blooms and the subsequent release of hepatotoxic microcystins (MCs) pose a serious threat to the safety of water for human and livestock consumption, agriculture irrigation, and aquaculture worldwide. Microcystin-LR (MC-LR), the most toxic variant of MCs, has been widely detected in a variety of environments such as water, sediments, plants, and many aquatic organisms. Conventional solutions of water treatment are costly, requiring specific infrastructure, as well as specialized personnel and equipment. Therefore, these solutions are not feasible in many rural areas or in the treatment of large reservoirs. In this regard, low-cost and low-technology solutions, such as constructed wetlands (CWs), are attractive solutions to treat surface waters contaminated with toxic cyanobacteria blooms from lakes, ponds, reservoirs, and irrigation systems. In line with this, the main aim of this work was to evaluate the potential of CWs for the treatment of water contaminated with MC-LR produced by Microcystis aeruginosa&mdash, LEGE 91094. For that, microcosms (0.4 &times, 0.3 &times, 0.3 m) simulating CWs were assembled with Phragmites australis to treat lake water contaminated with Microcystis aeruginosa cells and MCs. Results showed removal percentages of M. aeruginosa cells above 94% and about 99% removal of MC-LR during 1 week treatment cycles. CWs maintained their functions, regardless the presence of MC-LR in the system, and also showed significant removal of nutrients (ammonium ion removal up to 86%) and organic matter (removal reaching 98%). The present work indicates that CWs have the potential for removal of cyanobacterial cells and cyanotoxins, which can be useful for the treatment of eutrophic waters and provide water of sufficient quality to be used, for instance, in agriculture.

The data presented herein relates to the article entitled “Multibiomarker interactions to diagnose and follow-up chronic exposure of a marine crustacean to hazardous and noxious substances (HNS)” (Abreu et al., 2018). Multibiomarker approaches, including molecular, biochemical, physiological and behaviour parameters, are recognised as valuable and cost-effective to employ in integrated chemical and biological effects monitoring of aquatic contamination. Many biomarkers assessed in such programmes share common physiological pathways, showing concomitant or interdependent responses, which can reflect in increased energy costs related to physiological acclimation. Though, routine single biomarker data analysis, and exploratory principal component analysis, limit information obtained from the data collected and their functional interpretation. Ultimately, this influences the type of management actions taken to protect an affected ecosystem. This article presents data employed to develop an analytical approach accounting for multibiomarker interactions. The method was useful to diagnose and follow-up long-term exposure of the marine green crab (Carcinus maenas) to Hazardous and Noxious Substances (HNS). © 2018 The Authors This article is a result of INNOVMAR - Innovation and Sustainability in the Management and Exploitation of Marine Resources (reference NORTE-01-0145-FEDER-000035), ECOSERVICES, supported by the North Portugal Regional Operational Programme (NORTE2020), under the PORTUGAL2020 Partnership Agreement, through the European Regional Development Fund (ERDF). This work was partially funded by Strategic Funding UID/Multi/04423/2013 through the Portuguese Foundation for Science and Technology (FCT) and ERDF . Isabel Oliveira Abreu was supported by a FCT Ph.D grant (SFRH/BD/115529/2016). Transparency document

Hydrocarbon degradation (HD) potential by autochthonous microorganisms in the coastal sediments of the NW area of Portugal (coastal sandy beaches and estuaries of the rivers Minho and Douro) was evaluated, investigating if water and sediment contamination/characteristics influence it. Sediments were characterized for microbial abundance (by DAPI), HD microorganisms' abundance (by MPN), microbial community structure (by ARISA), hydrocarbons (by FTIR and SPME-GC-MS), hazardous and noxious substances (SPME-GC-MS) and metals (by AAS). To our knowledge, this is the first time all these pollutants, including the selected HNS, were measured simultaneously in sediments of the selected coastal area. Higher contaminants concentrations were, generally, registered in Douro samples. A clear differentiation of the microbial community structure between beaches and estuaries was observed, as well as, between Douro and Minho river estuaries. BIO-ENV analysis indicated both sediment characteristics (e.g. OM content) and contaminants presence/concentrations (e.g. tetrachloroethylene presence) affected the structure of the microbial community along the studied areas. In all the selected sites, the characterized autochthonous microbial communities showed potential for hydrocarbons degradation, with HD microorganisms being found in all collected sediments. These microorganisms can be a valuable asset to recover contaminated areas, but sediment characteristics and contaminants presence/levels need to be taken into account as they can affect their bioremediation potential and the success of their application as biotechnological tool. © 2018 Elsevier This research was partially supported by the Strategic Funding UID/Multi/04423/2013 through national funds provided by FCT – Foundation for Science and Technology and European Regional Development Fund (ERDF), in the framework of the programme PT2020, by the structured Program of R&D&I INNOVMAR - Innovation and Sustainability in the Management and Exploitation of Marine Resources, reference NORTE-01-0145-FEDER-000035 , namely within the Research Line ECOSERVICES (Assessing the environmental quality, vulnerability and risks for the sustainable management of the NW coast natural resources and ecosystem services in a changing world) within the R&D Institution CIIMAR (Interdisciplinary Centre of Marine and Environmental Research), supported by the Northern Regional Operational Programme ( NORTE2020 ), through the European Regional Development Fund (ERDF) and by the research project SpilLess - First line response to oil spills based on native microorganisms cooperation ( EASME/EMFF/2016/1.2.1.4/010 ) supported by the Executive Agency for Small and Medium-sized Enterprises (EASME) delegated by the European Commission. Appendix A

The aim of the present study was to test biomarker responses in an edible mollusk, Donax trunculus L. (Mollusca, Bivalvia) associated with environmental pollution in the Gulf of Annaba (northeastern Algeria). The biomarkers selected were glutathione S-transferase (GST), acetylcholinesterase (AChE) and metallothioneins (MTs). Samples were collected seasonally (September 2014, and January, April and July 2015) from two sites located over the Gulf of Annaba: El Battah and Sidi Salem. The results obtained reveal that autumn and winter were the two seasons that show an increase in GST activity, an inhibition of AChE activity and a high rate of MT. In addition, a decrease in AChE activity, an increase in both GST activity and MT levels in D. Trunculus collected from Sidi Salem in comparison with those of El Battah were observed. The biomarker responses at the Sidi Salem site reflect the presence of certain pro-oxidative compounds such as metals (Cd, Cu, Pb, Zn, Mn and Fe) determined in sediments in winter (January) 2015. Moreover, metal concentrations, except Fe, were higher at Sidi Salem than at El Battah. Overall, the Gulf of Annaba remains contaminated by heavy metal. However, this metallic contamination is relatively low and the risks for local population via this edible species were also low. © 2018 Académie des sciences This research was supported by the Algerian Fund for Scientific Research (Laboratory of Applied Animal Biology to Prof. N. Soltani), and by the Ministry of High Education and Scientific Research of Algeria (CNEPRU Project to Prof. N. Soltani) and by Centro Interdisciplinary de Investigação Marinha e Ambientol (CIIMAR), Portugal. Strategic Funding UID/Multi/04423/2013 through national funds provided by FCT–Foundation for Science and Technology and European Regional Development Fund (ERDF), in the framework of the program PT2020.

Marine pollution has been increasing as a consequence of anthropogenic activities. The preservation of marine ecosystems, as well as the safety of harvested seafood, are nowadays a global concern. Here, we report for the first time the contamination levels of a large set of 99 emerging and persistent organic contaminants (butyltins (BTs), polycyclic aromatic hydrocarbons (PAHs), pesticides including pyrethroids, pharmaceuticals and personal care products (PCPs) and flame retardants) in roe/gonads of sea urchin Paracentrotus lividus. Sea urchins are a highly prized worldwide delicacy, and the harvesting of this seafood has increased over the last decades, particularly in South West Atlantic coast, where this organism is harvested mainly for exportation. Sampling was performed in three harvesting sites of the NW Portuguese coast subjected to distinct anthropogenic pressures: Carreço, Praia Norte and Vila Chã, with sea urchins being collected in the north and south areas of each site. Butyltins and pharmaceuticals were not found at measurable levels. Several PAHs, four pyrethroids insecticides, four PCPs and eleven flame retardants were found in roe/gonads of sea urchins, though in general at low levels. Differences among harvesting sites and between areas within each site were found, the lowest levels of contaminants being registered in Carreço. The accumulation of contaminants in sea urchins’ roe/gonads seemed to reflect the low anthropogenic pressure felt in the sampling sites. Nevertheless, taking into account the low accumulated levels of chemicals, results indicate that sea urchins collected in South West Atlantic coast are safe for human consumption. © 2017 Elsevier Inc. Authors acknowledge INSEAFOOD colleagues that carried out sea urchin samplings. This work has been financially supported by the European project ‘Real time monitoring of SEA contaminants by an autonomous lab-on-a-chip biosensor ( SEA-on-a-CHIP ; No. 614168 ) and the research line INSEAFOOD within the Structured Program of R&D&I INNOVMAR – Innovation and Sustainability in the Management and Exploitation of Marine Resources (Reference NORTE-01–145-FEDER-000035 ) is funded by the Northern Regional Operational Programme ( NORTE2020 ) through the European Regional Development Fund (ERDF). António Marques acknowledge FCT for the IF2014 contract. Carolina Camacho acknowledges MARINALGAE4aqua Project ( ERA-NETCOFASP/004/2015 ) for the research grant. Part of this work was conducted in the Unit of Environmental, Organic and Biochemical high resolution analysis-Orbitrap-LC-MS analysis of the University of Ioannina, Greece and the authors would like to thank the Unit for providing access to the facilities.

Due to their ubiquitous presence, size and characteristics as ability to adsorb pollutants, microplastics are hypothesized as causing a major impact on smaller organisms, such as plankton. Despite this, there is a need to determine whether these impacts just relate to the environmental presence of the materials or their effects on biological processes. Therefore, we aimed to 1) review current research on plankton and microplastics; 2) compare field and laboratory experimental findings, and 3) identify knowledge gaps. The systematic review showed that 70% of the 147 relevant scientific publications were from laboratory studies and microplastics interactions with plankton were recorded in 88 taxa. Field study publications were relatively scarce and the characteristics of microplastics collected in the field were very different from those used in laboratory experiments thereby limiting the comparison between studies. Our systematic review highlighted knowledge gaps in: 1) the number of field studies; 2) the non-comparability between laboratory and field conditions, and 3) the low diversity of plankton species studied. Furthermore, this review indicated that while there are many studies on contamination by microplastics, the effects of this contamination (i.e., pollution per se) have been less well-studied, especially in the field at population, community, and ecosystem levels.

Ozone-based water treatments were investigated to simultaneously remove organic micropollutants (OMPs) and bacteria from surface water samples collected in a river reservoir supplying a drinking water treatment plant. Among the 21 environmentally relevant OMPs selected, 15 were detected at low ( O 3 L−1 and HRT: 10 min). Under these conditions, the removals of spiked OMPs were also higher than 85% (except for PFOS), whereas bacteria (i.e., total heterotrophs at 37 and 22 °C, E. coli and enterococci) decreased to values lower than the permissible levels for drinking water, even upon storage for 3-days. However, lower ozone doses (1.5 mg O 3 L−1) could not prevent the regrowth of total heterotrophs incubated at 37 °C, which reached levels five times higher than those recommended. Because drinking water treatments should not only consider OMPs abatement but also microbial inactivation for long periods of time, the results herein obtained point out the importance of using ozone doses capable of accomplish both targets when using single ozonation.

The presence of emergent antibiotics, in livestock wastewater may affect constructed wetlands (CWs) performance in the removal of other pollutants. The main objective of this study was to evaluate the influence of two antibiotics commonly used in livestock industry, enrofloxacin and ceftiofur, on metal removal by CWs. Microcosms (0.4 m×0.3 m×0.3 m), simulating CWs, were constructed with Phragmites australis to treat livestock wastewater spiked or not with 100 µg/L of enrofloxacin or ceftiofur (individually or in mixture). Wastewater was treated during 20 one-week cycles. After one-week cycle wastewater was removed and replaced by new wastewater (with or without spiking). At weeks 1, 2, 4, 8, 14, 18 and 20, treated wastewater was analysed to determine the removal rates of metals (Zn, Cu, Fe and Mn) and of each antibiotic. At weeks 1, 8 and 20 portions of the plant root substrate were collected and metals determined. At the end of the experiment metal levels were also determined in plant tissues. Removal rate of Fe from wastewater was 99%. Removal rates of Cu and Zn were higher than 85% and 89%, respectively, whereas for Mn removal rates up to 75% were obtained. In general, no significant differences were observed through time in the removals of the different metals, indicating that the systems maintained their functionality during the experimental period. Antibiotics did not interfere with the system depuration capacity, in terms of metals removals from wastewater, and ceftiofur even promoted metal uptake by P. australis. Therefore, CWs seem to be a valuable alternative to remove pollutants, including antibiotics and metals, from livestock wastewaters, reducing the risk the release of these wastewaters might pose into the environment, although more research should be conducted with other antibiotics in CWs. © 2016 Elsevier This research was partially supported by the Strategic Funding UID/Multi/04423/2013 through national funds provided by FCT – Foundation for Science and Technology and European Regional Development Fund (ERDF), in the framework of the programme PT2020 and by the structured Programme of R&D&I INNOVMAR - Innovation and Sustainability in the Management and Exploitation of Marine Resources, reference NORTE-01-0145-FEDER-000035, namely within the Research Line ECOSERVICES (Assessing the environmental quality, vulnerability and risks for the sustainable management of the NW coast natural resources and ecosystem services in a changing world) within the R&D Institution CIIMAR (Interdisciplinary Centre of Marine and Environmental Research), supported by the Northern Regional Operational Programme (NORTE2020), through the European Regional Development Fund (ERDF).

Contaminants of emerging concern (CEC) discharged in effluents of wastewater treatment plants (WWTPs), not specifically designed for their removal, pose serious hazards to human health and ecosystems. Their impact is of particular relevance to wastewater disposal and re-use in agricultural settings due to CEC uptake and accumulation in food crops and consequent diffusion into the food-chain. This is the reason why the chemical CEC discussed in this review have been selected considering, besides recalcitrance, frequency of detection and entity of potential hazards, their relevance for crop uptake. Antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs) have been included as microbial CEC because of the potential of secondary wastewater treatment to offer conditions favourable to the survival and proliferation of ARB, and dissemination of ARGs. Given the adverse effects of chemical and microbial CEC, their removal is being considered as an additional design criterion, which highlights the necessity of upgrading conventional WWTPs with more effective technologies. In this review, the performance of currently applied biological treatment methods for secondary treatment is analysed. To this end, technological solutions including conventional activated sludge (CAS), membrane bioreactors (MBRs), moving bed biofilm reactors (MBBRs), and nature-based solutions such as constructed wetlands (CWs) are compared for the achievable removal efficiencies of the selected CEC and their potential of acting as reservoirs of ARB&ARGs. With the aim of giving a picture of real systems, this review focuses on data from full-scale and pilot-scale plants treating real urban wastewater. To achieve an integrated assessment, technologies are compared considering also other relevant evaluation parameters such as investment and management costs, complexity of layout and management, present scale of application and need of a post-treatment. Comparison results allow the definition of design and operation strategies for the implementation of CEC removal in WWTPs, when agricultural reuse of effluents is planned.

Denitrification and anammox are key processes for reducing the external nitrogen loads delivered to coastal ecosystems, and these processes can be affected by pollutants. In this study, we investigated the effect of crude oil on denitrification and anammox. Controlled laboratory experiments were performed using sediment slurries from the Lima Estuary (NW Portugal). Anammox and denitrification rates were measured using 15N-labeled NO3−, and the production of 29N2 and 30N2 quantified by membrane inlet mass spectrometry. Results revealed that while denitrification rates were stimulated between 10 and 25 000 times after crude oil amendment, anammox activity was partially (between 2 and 5 times) or completely inhibited by the addition of crude oil when comparing to rates in unamended controls. Similar results were observed across four estuarine sediment types, despite their different physical-chemical characteristics. Moreover, N2O production was reduced by 2–36 times following crude oil addition. Further work is required to fully understand the mechanism(s) of the observed reduction in N2O production. This study represents one of the first contributions to the understanding of the impact of crude oil pollution on denitrification and anammox, with profound implications for the management of aquatic ecosystems regarding eutrophication (N-removal).

Metal exposure is known to induce the production and secretion of substances, such as aliphatic low molecular weight organic acids (ALMWOAs), into the rhizosphere by plant roots. Knowledge on this matter is extensive for soil plants but still considerably scarce regarding marsh plants roots adapted to high salinity media. Phragmites australis and Halimione portulacoides , two marsh plants commonly distributed in European estuarine salt marshes, were used to assess the response of roots of both species, in terms of ALMWOAs exudation, to Cu, Ni and Cd exposure (isolated and in mixture since in natural environment, they are exposed to mixture of metals). As previous studies were carried out in unrealistic and synthetic media, here a more natural medium was selected. Therefore, in vitro experiments were carried out, with specimens of both marsh plants, and in freshwater contaminated with two different Cu, Ni and Cd concentrations (individual metal and in mixture). Both marsh plants were capable of liberating ALMWOAs into the surrounding medium. Oxalic, citric and maleic acids were found in P. australis root exudate solutions and oxalic and maleic acids in H. portulacoides root exudate solutions. ALMWOA liberation by both plants was plant species and metal-dependent. For instance, Cu affected the exudation of oxalic acid by H. portulacoides and of oxalic and citric acids by P. australis roots. In contrast, Ni and Cd did not stimulate any specific response. Regarding the combination of all metals, H. portulacoides showed a similar response to that observed for Cu individually. However, in the P. australis case, at high metal concentration mixture, a synergetic effect led to the increase of oxalic acid levels in root exudate solution and to a decrease of citric acid liberation. A correlation between ALMWOAs exudation and metal accumulation could not be established. P. australis and H. portulacoides are considered suitable metal phytoremediators of estuarine impacted areas. Understanding the mechanisms developed by these plants which allow them to tolerate and remediate metal-contaminated sediments is important to potentiate their use in phytoremediation purpose. This work provides new knowledge regarding the H. portulacoides and P. australis ability to exude ALMWOAs in response to metal contamination.

Oil spill pollution remains a serious concern in marine environments and the development of effective oil bioremediation techniques are vital. This work is aimed at developing an autochthonous hydrocarbon-degrading consortium with bacterial strains with high potential for hydrocarbons degradation, optimizing first the growth conditions for the consortium, and then testing its hydrocarbon-degrading performance in microcosm bioremediation experiments. Bacterial strains, previously isolated from a sediment and cryopreserved in a georeferenced microbial bank, belonged to the genera Pseudomonas, Rhodococcus and Acinetobacter. Microcosms were assembled with natural seawater and petroleum, for testing: natural attenuation (NA), biostimulation (BS) (nutrients addition), bioaugmentation with inoculum pre-grown in petroleum (BA/P) and bioaugmentation with inoculum pre-grown in acetate (BA/A). After 15 days, a clear blending of petroleum with seawater was observed in BS, BA/P and BA/A but not in NA. Acetate was the best substrate for consortium growth. BA/A showed the highest hydrocarbons degradation (66%). All bacterial strains added as inoculum were recovered at the end of the experiment. This study provides an insight into the capacity of autochthonous communities to degrade hydrocarbons and on the use of alternative carbon sources for bacterial biomass growth for the development of bioremediation products to respond to oil spills.

Floating wetland islands (FWI) are considered nature-based solutions with great potential to promote several ecosystem services, such as biodiversity and water quality enhancement through phytoremediation processes. To our knowledge, the present work is the first to scientifically document the in-situ establishment of an FWI in a seawater port marina. The establishment and performance of a cork floating platform with a polyculture (Sarcocornia perennis, Juncus maritimus, Phragmites australis, Halimione portulacoides, Spartina maritima, Limonium vulgare) was evaluated. The diversity of organisms present in the FWI was undertaken based on the macrofauna assessment, taking into consideration marine water characterization, with a focus on hydrocarbons. Microbial communities were assessed based on metabarcoding approach to study 16S rRNA gene from environmental DNA retrieved from biofilm (from the planting media), marine biofouling (from the submerged platform) and surface marina water. S. perennis was the species with the highest survival rate and growth. The structure of the microbial community showed clear differences between those established in the FWI and those in the surrounding water, showing the presence of some bacterial groups that can be relevant for bioremediation processes (e.g., Saprospiraceae family). Concerning the macrofauna analysis, Mytilus sp. was the predominant taxa. To be of relevance, total petroleum hydrocarbons were detected at the marina up to ca. 6 mg/L. This study gives new insights into broadening FWI application to the saline environments of port marinas and to supporting a management strategy to promote several ecosystem services such biodiversity, species habitat, water quality enhancement and added aesthetic value to the marina landscape.

This study aimed to investigate if microplastics (MPs) type (polyethylene microspheres (mPE), fishing line fibers, film plastic bags MPs and bottle cap particles) and aging affect MPs capacity to sorb Cd or Cu in estuarine salt marsh medium. Tests were carried out in elutriate solution, a simple medium obtained by mixing rhizosediment (sediment in contact with plants roots) with the respective estuarine water, that can be used to simulate water-sediment exchanges in estuarine salt marsh environments. After 7 days of exposure, metals adsorption was only detected for film MPs. No differences were observed between virgin and aged MPs. Salinity also did not influence metal adsorption to mPE. Present results indicate that in estuarine salt marsh areas some types of MPs might adsorb metals, which could affect metals availability.

The study aim was to determine a range of relevant persistent and emerging pollutants in oysters produced in an aquaculture facility located in an important production area, to assure their safety for human consumption. Pollutants, including 16 PAHs, 3 butyltins (BTs), 29 flame retardants (FRs, including organophosphate and halogenated FRs), 35 pesticides (including 9 pyrethroid insecticides) and 13 personal care products (PCPs, including musks and UV filters), were determined in oysters' tissues collected during one year in four seasonal sampling surveys. The seasonal environmental pollution on the production site was evaluated by water and sediment analysis. Furthermore, oysters' nutritional quality was also assessed and related with the consumption of healthy seafood, showing that oysters are a rich source of protein with low fat content and with a high quality index all year around. Results showed that most analysed pollutants were not detected either in oyster tissues or in environmental matrixes (water and sediments). The few pollutants detected in oyster tissues, including both regulated and non-legislated pollutants, such as a few PAHs (fluorene, phenanthrene, anthracene, fluoranthene, pyrene and indenopyrene), FRs (TPPO, TDCPP, DCP, BDE-47, BDE-209 and Dec 602) and PCPs (galaxolide, galaxolidone, homosalate and octocrylene), were present at low levels (in the ng/g dw range) and did not represent a significant health risk to humans. The observed seasonal variations related to human activities (e.g. tourism in summer) highlights the need for environmental protection and sustainable resource exploration for safe seafood production. © 2019 Elsevier B.V., This work was supported by the Structured R&D&I Project INNOVMAR – “Innovation and Sustainability in the Management and Exploitation of Marine Resources” (ref. NORTE-01-0145-FEDER-000035) within the research line “INSEAFOOD - Innovation and valorization of seafood products: meeting local challenges and opportunities”, founded by the Northern Regional Operational Programme (NORTE2020) through the European Regional Development Fund (ERDF). This study was also supported by the SEA-on-a-CHIP project, funded from European Union Seventh Framework Programme (FP7-OCEAN-2013) under grant agreement No. 614168. This work has been also financially supported by the Generalitat de Catalunya (Consolidated Research Group 2017 SGR 01404 – Water and Soil Quality Unit). Biotage is acknowledged for providing SPE cartridges. Authors acknowledge Carlos R. Gomes for helping on multivariated analysis.

The present work aimed to explore the potential of autochthonous microorganisms from an urban estuary and from activated sludge of an associated wastewater treatment plant (WWTP), for biodegradation of an antidepressant drug, paroxetine, and on a cholesterol-lowering agent, bezafibrate. These compounds were chosen as representatives of extensively used pharmaceuticals. Autochthonous microorganisms from the indicated sources were exposed to the target pharmaceuticals (1 mg/L) in co-metabolism with sodium acetate (500 mg/L) along a two-weeks period, for a total of 7 two-weeks periods (here referred as cycles). Exposures were carried out in batch mode, under different incubation conditions (agitation vs. static). Removal of pharmaceuticals was monitored at the end of each cycle, by analysing the culture medium. For paroxetine, fluoride ion release was also followed as an indicator of defluorination of the molecule. The structure of the bacterial communities was analysed by ARISA (Automated rRNA Intergenic Spacer Analysis), at the beginning of the experiment and at the end of the first and the last cycles to identify substantial changes associated with the time of exposure, the incubation conditions and the presence and type of pharmaceuticals. Incubation conditions affected not only the bacterial community structure, but also the biodegradation efficiency. At the beginning of the experiment, removal of target pharmaceuticals was found to be lower under agitation than under static conditions, but at the end of the experiment, results showed high removal of the pharmaceuticals from the culture medium (>97%) under both conditions, mainly by microbiological processes. For paroxetine, adsorption and abiotic processes also had an important influence on its removal, but defluorination only occurred in the presence of microorganisms. These results highlight that autochthonous microorganisms from estuarine sediments and WWTP sludge have high ability to remove the selected pharmaceuticals with relevant implications for the development of new bioremediation tools for environmental restoration.

Integrated compensatory responses of physiological systems towards homeostasis are generally overlooked when it comes to analysing alterations in biochemical parameters indicative of such processes. Here an hypothesis-driven multivariate analysis accounting for interactive multibiomarker responses was used to investigate effects of long-term exposure of Carcinus maenas to Hazardous and Noxious Substances (HNS). Adult male crabs were exposed to low and high post-spill levels of acrylonitrile (ACN) or aniline (ANL) for 21d. Bioaccumulation, feeding behaviour, and biomarkers related to mode-of-action (MoA) (detoxification, neurotransmission and energy production) were evaluated over time. Distinct temporal patterns of response to low and high exposure concentrations were depicted, with a main set of interactive multibiomarker predictors identified for each HNS (five for ACN and three for ANL), useful to follow coupled evolvement of biomarker responses. ACN caused peripheral neurotoxic effects coupled with enhanced biotransformation and significant oxidative damage particularly relevant in gills. ANL elicited alterations in central neurotransmission affecting ventilation coupled with very low levels of oxidative damage in gills. Results indicate chronic toxicity data are determinant to improve HNS hazard assessment if the aim is to obtain reliable risk calculations, and develop effective predictive models avoiding overestimation but sufficiently protective. Accounting for multibiomarker interactions brought otherwise overlooked information about C. maenas responses and MoA of ACN and ANL. Accounting for biomarker interactions drastically improves the quality of the model to diagnose toxic effects. © 2018 Elsevier This article is a result of INNOVMAR - Innovation and Sustainability in the Management and Exploitation of Marine Resources (reference NORTE-01-0145-FEDER-000035), ECOSERVICES, supported by NORTE2020, PORTUGAL2020 Partnership Agreement, through ERDF . This work was partially funded by Strategic Funding UID/Multi/04423/2013 through FCT and ERDF .

Constructed wetlands (CWs) can be used to reduce various pollutants present in livestock wastewater, such as organic matter, nutrients and metals. Very recently these systems have also been used to remove the so called emergent pollutants. These pollutants can be harmful for both microorganisms and plants, two key players in CWs removal processes. Therefore, the aim of the present work was to assess the influence of emergent pollutants, namely antibiotics, on the removal of pollutants from livestock wastewaters, as antibiotics might decrease CWs performance for the treatment of this type of wastewater. Microcosms (0.4 m × 0.3 m × 0.3 m), simulating CWs, were assembled with Phragmites australis to treat livestock wastewater not doped or doped with 100 μg/L of enrofloxacin and/or of ceftiofur, two antibiotics commonly used in livestock industry. Four different treatments (Control, Enr, Cef and Mix) were tested, each in triplicate. Wastewater was treated during one-week cycle, after which it was removed and replaced by new wastewater (doped or not), in a total of 8 cycles. At weeks 1, 2, 4 and 8 treated wastewater was collected and analysed to determine removal rates of nutrients (ammonia, nitrate, nitrite and phosphate), organic matter (chemical oxygen demand (COD) and biological oxygen demand (BOD)), and solids (including total suspended solids (TSS)), as well as, veterinary antibiotics (enrofloxacin and ceftiofur). High removal rates (up to 90% depending of the parameter) were observed independently of the presence of the veterinary antibiotics, which were also significantly removed from the wastewater. Generally, measured parameters presented values lower than those expressed in the legislation for wastewater discharge into the aquatic environment. Present results indicate that, in tested conditions, the presence of veterinary antibiotics, namely enrofloxacin and ceftiofur, did not influence significantly the biochemical removal processes that occur naturally in CWs during treatment of livestock wastewater, the systems maintaining their performance. Therefore, CWs are a valuable alternative to remove pollutants, including antibiotics, from livestock wastewaters, reducing the impact of this type of effluents into the environment. In addition, this technology can be an efficient/economically viable technology to meet the current wastewater reuse challenges. © 2017 Elsevier This research was partially supported by the Strategic Funding UID/Multi/04423/2013 through national funds provided by FCT – Foundation for Science and Technology and European Regional Development Fund (ERDF), in the framework of the programme PT2020 and by the structured Program of R&D&I INNOVMAR – Innovation and Sustainability in the Management and Exploitation of Marine Resources, reference NORTE-01-0145-FEDER-000035, namely within the Research Line ECOSERVICES (Assessing the environmental quality, vulnerability and risks for the sustainable management of the NW coast natural resources and ecosystem services in a changing world) within the R&D Institution CIIMAR (Interdisciplinary Centre of Marine and Environmental Research), supported by the Northern Regional Operational Programme (NORTE2020), through the European Regional Development Fund (ERDF).

Autochthonous bioaugmentation for metal phytoremediation is still little explored, particularly its application to estuarine salt marshes, but results obtained so far are promising. Nevertheless, understanding the behaviour of the microbial communities in the process of bioaugmentation and their role in improving metal phytoremediation is very important to fully validate the application of this biological technology. This study aimed to characterize the bacterial community dynamic associated with the application of autochthonous bioaugmentation in an experimentation which showed that Phragmites australis rhizosphere microorganisms could increase this salt marsh plant potential to phytoremediate Cu contaminated sediments. Bacterial communities present in the autochthonous microbial consortium resistant to Cu added to the medium and in the sediment at the beginning and at the end of the experiment were characterized by ARISA. Complementarily, the consortium and the sediment used for its production were characterized by next generation sequencing using the pyrosequencing platform 454. The microbial consortium resistant to Cu obtained from non-vegetated sediment was dominated by the genus Lactococcus (46%), Raoultella (25%), Bacillus (12%) and Acinetobacter (11%), whereas the one obtained form rhizosediment was dominated by the genus Gluconacetobacter (77%), Bacillus (17%) and Dyella (3%). Results clearly showed that, after two months of experiment, Cu caused a shift in the bacterial community structure of sediments, an effect that was observed either with or without addition of the metal resistant microbial consortium. Therefore, bioaugmentation application improved the process of phytoremediation (metal translocation by the plant was increased) without inducing long term changes in the bacterial community structure of the sediments. So, phytoremediation combined with autochthonous bioaugmentation can be a suitable technology for the recovery of estuarine areas, contributing for an efficient risk management strategy of these coastal zones. © 2017 To Rayra Santiago, Tatiana Necrasov and Catarina Magalhães for their help in the experiments assembling and dismantling of the vessels and to Paula Rodrigues for helping on samples preparation and metal analysis. This research was partially supported by the Strategic Funding UID/Multi/04423/2013 through national funds provided by FCT – Foundation for Science and Technology and European Regional Development Fund (ERDF) , in the framework of the programme PT2020 and by the structured Program of R&D&I INNOVMAR - Innovation and Sustainability in the Management and Exploitation of Marine Resources , reference NORTE-01-0145-FEDER-000035 , namely within the Research Line ECOSERVICES (Assessing the environmental quality, vulnerability and risks for the sustainable management of the NW coast natural resources and ecosystem services in a changing world) within the R&D Institution CIIMAR (Interdisciplinary Centre of Marine and Environmental Research), supported by the Northern Regional Operational Programme ( NORTE2020 ), through the European Regional Development Fund (ERDF) .

This study aimed to investigate Cu oxide nanoparticles (CuO NP) effect on microbial communities associated with salt marsh plants (Halimione portulacoides and Pragmites australis) rhizosphere and its implications for phytoremediation processes. Experiments were conducted, under controlled conditions, over one week. Rhizosediment soaked in the respective elutriate (a simplified natural medium) with or without plants, was doped with CuO NP or with Cu in ionic form. Microbial community in rhizosediments was characterized in terms of abundance (by DAPI) and structure (by ARISA). Metal uptake by plants was evaluated by measuring Cu in plant tissues (by atomic absorption spectroscopy). Results revealed significant metal uptake but only in plant roots, which was significantly lower (H. portulacoides) or not significant (P. australis) when the metal was in NP form. Microbial community structure was significantly changed by the treatment (absence/presence of Cu, ionic Cu or CuO NP) as showed by multivariate analysis of ARISA profiles and confirmed by analysis of similarities (Global test - one way ANOSIM). Moreover, in P. australis rhizosediments microbial abundance, bacterial richness and diversity indexes were significantly affected (increased or decreased) due to metal presence whereas in H. portulacoides rhizosediment microbial abundance showed a significant decrease, particularly when the metal was in NP form. Accordingly, Cu presence affected the response of the rhizosphere microbial community and in some cases that response was significantly different when Cu was in NP form. The response of the microbial communities to Cu NP might also contribute to the lower metal accumulation by plants when the metal was in this form. So, Cu NP may cause disturbances in ecosystems functions, ultimately affecting phytoremediation processes. These facts should be considered regarding the use of appropriate salt marshes plants to remediate moderately impacted areas such as estuaries, where NPs can be found. © 2017 Elsevier To C?tia Caetano and Paula Rodrigues for their help in experimental assembling and metal determination. This research was partially supported by the Strategic Funding UID/Multi/04423/2013 through national funds provided by FCT ? Foundation for Science and Technology and European Regional Development Fund (ERDF), in the framework of the programme PT2020 and by the structured Program of R&D&I INNOVMAR - Innovation and Sustainability in the Management and Exploitation of Marine Resources, reference NORTE-01-0145-FEDER-000035, namely within the Research Line ECOSERVICES (Assessing the environmental quality, vulnerability and risks for the sustainable management of the NW coast natural resources and ecosystem services in a changing world) within the R&D Institution CIIMAR (Interdisciplinary Centre of Marine and Environmental Research), supported by the Northern Regional Operational Programme (NORTE2020), through the European Regional Development Fund (ERDF). Authors acknowledge also FCT for the PhD scholarship SFRH/BD/112154/2015.

Constructed wetlands (CWs) are defined as engineered wetlands that utilize natural processes involving soil, wetland vegetation and their associated microbial assemblages to assist the treatment of wastewaters or other polluted water sources. Although CWs have been applied to different types of wastewaters, CWs application to livestock wastewaters is more complicated do to the characteristics of these waters and only a few studies have reported CWs application in these cases. Livestock wastewater can contain diverse veterinary drugs, including antibiotics, which are normally not removed in wastewater treatment plants. Consequently, veterinary antibiotics or their active compounds can enter directly in the water system through effluent discharges, which can lead to serious toxic effects in organisms and promote antibiotic resistance. Therefore, efficient wastewater treatments are needed. Considering the problematic of antibiotics release in the environment, the need for methodologies to efficiently remove these compounds from wastewater effluents, namely from livestock wastewater effluents, and the scarcity of studies on the application of CWs to deal with this problem, authors have been developing a series of studies to evaluated CWs applicability for livestock wastewater treatment. Studies have been assessing not only antibiotics removal but also antibiotic possible effects on CWs performance. These studies will be presented in this chapter.

91 Páginas, 4 Tablas, 6 Figuras, Optimal supply of trace elements (TE) is a prerequisite for microbial growth and activity in anaerobic digestion (AD) bioprocesses. However, the required concentrations and ratios of essential TE for AD biotechnologies strongly depend on prevailing operating conditions as well as feedstock composition. Furthermore, TE in AD bioreactors undergo complex physico-chemical reactions and may be present as free ions, complex bound or as precipitates depending on pH, or on the presence of sulfur compounds or organic macromolecules. To overcome TE deficiency various commercial mineral products are typically applied to AD processes. The addition of heavy metals poses the risk of overdosing operating systems, which may be toxic to microbial consortia and ultimately the environment. Adequate supplementation, therefore, requires not only appropriate knowledge about the composition, but also on the speciation and bioavailability of TE. However, very little is yet fully understood on this specific issue. Evaluations of TE typically only include the measurement of total TE concentrations but do not consider the chemical forms in which TE exist. Thus detailed information on bioavailability and potential toxicity cannot be provided. This review provides an overview of the state-of-the-art in approaches to determine bioavailable TE in anaerobic bioprocesses, including sequential fractionation and speciation techniques. Critical aspects and considerations, including with respect to sampling and analytical procedures, as well as mathematical modelling, are examined. The approaches discussed in this review are based on our experiences and on previously published studies in the context of the ‘COST Action 1302: European Network on Ecological Roles of Trace Metals in Anaerobic Biotechnologies'., This article is based upon work from COST Action 1302 (‘European Network on Ecological Roles of Trace Metals in Anaerobic Biotechnologies’) supported by COST (European Cooperation in Science and Technology). EvH and GE acknowledge support from the European Union's Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No. 643071 (‘Advanced Biological Waste-to-Energy Technologies – ABWET’). GC acknowledges funding from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme grant agreement No. 261330. The authors thank the two anonymous reviewers for their comments, which significantly improved the quality of the manuscript.

This study aimed to evaluate the response of the microbial community from CWs microcosms tested for the removal of two veterinary antibiotics, enrofloxacin (ENR) and tetracycline (TET), from livestock industry wastewater. Three treatments were tested (control, ENR or TET (100 μg L −1 )) over 12 weeks in microcosms unplanted and planted with Phragmites australis . CWs removal efficiency was relatively stable along time, with removals higher than 98% for ENR and 94% for TET. In addition, CWs were able to reduce wastewater toxicity, independently of antibiotics presence. Despite no significant differences were observed in terms of microbial abundance, bacterial richness or diversity, analysis of similarities (two-way crossed ANOSIM) showed a significant effect of both time and treatments in bacterial community structure. This study points to CWs applicability for veterinary antibiotics removal from livestock wastewaters, showing that CWs microbial communities were able to adapt without significant changes in their diversity or depuration capacity.

Recent findings indicate that different Endocrine Disrupting Chemicals (EDCs) interfere with lipid metabolic pathways in mammals and promote fat accumulation, a previously unknown site of action for these compounds. The antifoulant and environmental pollutant tributyltin (TBT), which causes imposex in gastropod snails, induces an “obesogenic” phenotype in mammals, through the activation of the nuclear receptors retinoid X receptor (RXR) and peroxisome proliferator-activated receptor gamma (PPARγ). In teleosts, the effects of TBT on the lipid metabolism are poorly understood, particularly following exposure to low, environmental concentrations. In this context, the present work shows that exposure of zebrafish to 10 and 50 ng/L of TBT (as Sn) from pre-hatch to 9 months of age alters the body weight, condition factor, hepatosomatic index and hepatic triglycerides in a gender and dose related manner. Furthermore, TBT modulated the transcription of key lipid regulating factors and enzymes involved in adipogenesis, lipogenesis, glucocorticoid metabolism, growth and development in the brain and liver of exposed fish, revealing sexual dimorphic effects in the latter. Overall, the present study shows that the model mammalian obesogen TBT interferes with triglyceride accumulation and the transcriptional regulation of lipid metabolism in zebrafish and indentifies the brain lipogenic transcription profile of fish as a new target of this compound.

International audience; The potential effects of the mineralogical composition of sediment on the degradation of oil buried on sandy beaches were investigated. Toward that purpose, a laboratory experiment was carried out with sandy sediment collected along NW Iberian Peninsula beaches, tar-balls from the Prestige oil spill (NW Spain) and seawater. The results indicate that the mineralogical composition is important for the physical appearance of the oil (tar-balls or oil coatings). This finding prompted a reassessment of the current sequence of degradation for buried oil based on compositional factors. Moreover, the halo development of the oil coatings might be enhanced by the carbonate concentration of the sand. These findings open new prospects for future monitoring and management programs for oiled sandy beaches.

The presence of polycyclic aromatic hydrocarbons (PAHs) may change the mechanisms of metal uptake, thus influencing kinetics and extent of metal phytoextraction. Studies on the subject are scarce, particularly for salt marsh plants. The aim of this work was to investigate the effect of PAHs on the uptake of Cu by Halimione portulacoides, a plant commonly found in salt marshes. Experiments were carried out in the laboratory, either in hydroponics (sediment elutriate) or in sediment soaked in elutriate, which were prepared with sediment and water from a salt marsh of the Cavado river estuary (NW Portugal). Groups of H. portulacoides (grown in a greenhouse) were exposed to those media during six days. Cu2+ (as Cu(NO3)2), 10(2) and 10(4) microg l(-1), was added to the media as well as 1.6 microg l(-1) of the sixteen EPA priority PAHs (0.1 microg l(-1) of each PAHs). Cu was assayed in solutions, sediments and different plant tissues before and after experiments. After exposure, photosynthetic efficiency and levels of chlorophylls were also measured, indicating that plant stress indicators were identical in all plants independently of the media to which the plants were exposed. PAHs influenced both the soluble Cu fraction and Cu uptake by plants. The amounts of metal accumulated in both roots and stems were significantly higher when the 10(4) microg l(-1) of Cu enriched elutriate was amended with PAHs. Thus, results suggest that PAHs may modify Cu solubility, the Cu sorption by plants and/or the passive penetration of Cu into the root cells. Therefore, the combined effects of different types of pollutants should be taken in consideration when studying the remediation potential of plants, namely in terms of phytoextraction.

Fluoroquinolones and cephalosporins are two classes of veterinary antibiotics arising as pollutants of emerging concern. In this work, the microbial degradation of two representative antibiotics of both these classes, enrofloxacin (ENR) and ceftiofur (CEF), is reported. Biodegradation of the target antibiotics was investigated by supplementing the culture medium with ENR and CEF, individually and in mixture. Microbial inocula were obtained from rhizosphere sediments of plants derived from experimental constructed wetlands designed for the treatment of livestock wastewaters contaminated with trace amounts of these antibiotics. Selected microbial inocula were acclimated during a period of 5 months, where the antibiotics were supplemented every three weeks at the concentration of 1 mg L− 1, using acetate as a co-substrate. After this period, the acclimated consortia were investigated for their capacity to biodegrade 2 and 3 mg L− 1 of ENR and CEF. Complete removal of CEF from the inoculated culture medium was always observed within 21 days, independently of its concentration or the concomitant presence of ENR. Biodegradation of ENR decreased with the increase in its concentration in the culture medium, with defluorination percentages decreasing from ca. 65 to 4%. Ciprofloxacin and norfloxacin were detected as biodegradation intermediates of ENR in the microbial cultures supplemented with this antibiotic, indicating that defluorination of at least part of ENR in these cultures is not an immediate catabolic step. Abiotic mechanisms showed high influence in the removal of CEF, affecting less ENR degradation. The acclimation process with the target antibiotics led to significant shifts in the structure and diversity of the microbial communities, predominantly selecting microorganisms belonging to the phyla Proteobacteria (e.g. Achromobacter, Variovorax and Stenotrophomonas genera) and Bacteroidetes (e.g. Dysgonomonas, Flavobacterium and Chryseobacterium genera). The results presented in this study indicate that biodegradation can be an important mechanism for the environmental removal of the tested compounds. © 2016 Elsevier M.F. Carvalho wishes to acknowledge Investigator FCT program supported by Fundação para a Ciência e a Tecnologia (FCT) (IF/00791/2013), Fundo Social Europeu and Programa Operacional Potencial Humano. W. Gao and Z. Jia acknowledge the Strategic Priority Research Program of the Chinese Academy of Sciences (XDB15040000). This work was implemented in the framework of the structured program of R&D&I INNOVMAR - Innovation and Sustainability in the Management and Exploitation of Marine Resources (reference NORTE-01-0145-FEDER-000035), namely within the research line ECOSERVICES, supported by the Northern Regional Operational Programme (NORTE2020), through the European Regional Development Fund (ERDF).

This study investigated the uptake of silver nanoparticles (AgNPs) by a salt marsh plant, Phragmites australis, as well as AgNPs effects on rhizospheric microbial community, evaluating the implications for phytoremediation processes. Experiments were carried out with elutriate solution doped with Ag, either in ionic form or in NP form. Metal uptake was evaluated in plant tissues, elutriate solutions and sediments (by AAS) and microbial community was characterized in terms of bacterial community structure (evaluated by ARISA). Results showed Ag accumulation but only in plant belowground tissues and only in the absence of rhizosediment, the presence of sediment reducing Ag availability. But in plant roots Ag accumulation was higher when Ag was in NP form. Multivariate analysis of ARISA profiles showed significant effect of the absence/presence of Ag either in ionic or NP form on microbial community structure, although without significant differences among bacterial richness and diversity. Overall, P. australis can be useful for phytoremediation of medium contaminated with Ag, including with AgNPs. However, the presence of Ag in either forms affected the microbial community structure, which may cause disturbances in ecosystems function and compromise phytoremediation processes. Such considerations need to be address regarding environmental management strategies applied to the very important estuarine areas. Capsule The form in which the metal was added affected metal uptake by Phragmites australis and rhizosediment microbial community structure, which can affect phytoremediation. © 2017 Elsevier This research was partially supported by the Strategic Funding UID/Multi/04423/2013 through national funds provided by FCT – Foundation for Science and Technology and European Regional Development Fund (ERDF), in the framework of the programme PT2020 and by the structured Program of R&D&I INNOVMAR - Innovation and Sustainability in the Management and Exploitation of Marine Resources, reference NORTE-01-0145-FEDER-000035, namely within the Research Line ECOSERVICES (Assessing the environmental quality, vulnerability and risks for the sustainable management of the NW coast natural resources and ecosystem services in a changing world) within the R&D Institution CIIMAR (Interdisciplinary Centre of Marine and Environmental Research), supported by the Northern Regional Operational Programme (NORTE2020), through the European Regional Development Fund (ERDF). Authors acknowledge FCT for the PhD scholarship SFRH/BD/112154/2015.

Sea urchin represents one of the most valuable seafood product being harvested and explored for their edible part, the gonads or roe. This species is generally considered a sentinel organism for ecotoxicological studies being widely used in monitoring programs to assess coastal aquatic environments quality, because is directly exposed to anthropogenic contaminants in their habitat. In this context, the aim of this study is to evaluate the concentrations of macro (Cl, K, P, Ca, S) and trace (Zn, Br, Fe, Sr, I, Se, Rb, Cu, Cr, Ni, As, iAs, Cd, Pb, Hg) elements in Paracentrotus lividus gonads from three South West Atlantic production areas subjected to distinct environmental and anthropogenic pressures. In all studied areas, the elements profile in sea urchin gonads was Cl > K > P > Ca > S > Zn > Br > Fe > Sr > I > Rb > Cu > Se > Cr > Ni, suggesting an element guide profile with special interest for sea urchin farming development. Concerning toxic elements, the profile was the following: As > Cd > Pb > Hg > iAs. The results evidenced higher levels of Pb and Hg in open areas. Distinct area characteristics and anthropogenic pressures of production areas evidence the importance of biomonitoring contaminants, particularly toxic elements. In general, the levels of these elements were below maximum levels in foodstuffs (MLs) which pose a minimal health risk to consumers. © 2018 Elsevier Inc. This work was supported by the research line INSEAFOOD within the Structured Program of R&D&I INNOVMAR – Innovation and Sustainability in the Management and Exploitation of Marine Resources (Ref. NORTE-01-145-FEDER-000035 ) that is funded by the Northern Regional Operational Programme (NORTE2020) through the European Regional Development Fund (ERDF). The Portuguese Foundation for Science and Technology (FCT) supported the contract of António Marques in the framework of IF2014 program, as well as the post-doc Grant of Patrícia Anacleto ( SFRH/BPD/100728/2014 ). Carolina Camacho acknowledges MARINALGAE4aqua Project ( ERA-NET COFASP/004/2015 ) for the research grant.

The presence of organic micropollutants (MPs) in the aquatic environment is strongly related to their difficult elimination by conventional water and wastewater treatment processes. Therefore, alternative treatment technologies are required to overcome this problem. In this domain, constructed wetlands (CWs) have gained increasing attention in the last years, mainly due to the low-cost, simple operation/maintenance and environmental friendliness of these systems. However, studies on the application of CWs to remove MPs from freshwater aquaculture effluents are still scarce. In this work, planted (Phragmites australis) vertical subsurface flow CWs, at microcosm scale, were investigated for the removal of MPs found in non-spiked freshwater aquaculture effluents, namely atrazine, isoproturon, perfluorooctanesulfonic acid (PFOS), clarithromycin, erythromycin, fluoxetine, norfluoxetine, and 2-ethylhexyl-4-methoxycinnamate (EHMC). A wider multi-component set of 36 MPs was also studied by adding these MPs at 100 ng L −1 to the same matrix (alachlor, atrazine, chlorfenvinphos, isoproturon, PFOS, azithromycin, clarithromycin, erythromycin, diclofenac, methiocarb, acetamiprid, clothianidin, thiacloprid, thiamethoxam, EHMC, simazine, atorvastatin, bezafibrate, carbamazepine, cephalexin, ceftiofur, citalopram, clindamycin, clofibric acid, diphenhydramine, enrofloxacin, fluoxetine, ketoprofen, metoprolol, norfluoxetine, ofloxacin, propranolol, tramadol, trimethoprim, venlafaxine, and warfarin). High weekly removal efficiencies (>87%) were observed for all MPs in both non-spiked and spiked experiments, with the exception of EHMC (removal rates between 0 and 86%). These results emphasize the potential of CWs to remove MPs from freshwater aquaculture effluents, but also the need to enhance the performance of these systems for the elimination of some recalcitrant MPs, such as EHMC, which was found at high concentrations in the studied effluents. © 2018 Elsevier This work was financially supported by Project POCI-01-0145-FEDER-006984 – Associate Laboratory LSRE-LCM funded by ERDF (European Regional Development Fund) through COMPETE2020 - Programa Operacional Competitividade e Internacionalização (POCI) – and by national funds through Fundação para a Ciência e a Tecnologia (FCT). This research was also partially supported by CIIMAR (Interdisciplinary Centre of Marine and Environmental Research) Strategic Funding UID/Multi/04423/2013 through national funds provided by FCT and ERDF, in the framework of the programme PT2020 and by the structured Program of R&D&I INNOVMAR - Innovation and Sustainability in the Management and Exploitation of Marine Resources, reference NORTE-01-0145-FEDER-000035, namely within the Research Line INSEAFOOD within the R&D Institution CIIMAR, supported by the Northern Regional Operational Programme (NORTE2020), through the ERDF. AMG and ARR acknowledge the research grant from FCT (Refs. SFRH/BD/133117/2017 and SFRH/BPD/101703/2014, respectively). The authors would like to acknowledge the financial support provided by COST- European Cooperation in Science and Technology , to the COST Action ES1403: New and emerging challenges and opportunities in wastewater reuse (NEREUS). Disclaimer: The content of this article is the authors' responsibility and neither COST nor any person acting on its behalf is responsible for the use, which might be made of the information contained in it. Appendix A

This study investigates the potential of an indigenous estuarine microbial consortium to degrade two polycyclic aromatic hydrocarbons (PAHs), naphthalene and fluoranthene, under nitrate-reducing conditions. Two physicochemically diverse sediment samples from the Lima Estuary (Portugal) were spiked individually with 25 mg L−1 of each PAH in laboratory designed microcosms. Sediments without PAHs and autoclaved sediments spiked with PAHs were run in parallel. Destructive sampling at the beginning and after 3, 6, 12, 30 and 63 weeks incubation was performed. Naphthalene and fluoranthene levels decreased over time with distinct degradation dynamics varying with sediment type. Next-generation sequencing (NGS) of 16 S rRNA gene amplicons revealed that the sediment type and incubation time were the main drivers influencing the microbial community structure rather than the impact of PAH amendments. Predicted microbial functional analyses revealed clear shifts and interrelationships between genes involved in anaerobic and aerobic degradation of PAHs and in the dissimilatory nitrate-reducing pathways (denitrification and dissimilatory nitrate reduction to ammonium - DNRA). These findings reinforced by clear biogeochemical denitrification signals (NO3 − consumption, and NH4 + increased during the incubation period), suggest that naphthalene and fluoranthene degradation may be coupled with denitrification and DNRA metabolism. The results of this study contribute to the understanding of the dissimilatory nitrate-reducing pathways and help uncover their involvement in degradation of PAHs, which will be crucial for directing remediation strategies of PAH-contaminated anoxic sediments. © 2018 Elsevier The authors acknowledge the reviewers for their valuable comments and suggestions, which were helpful in improving the manuscript. This research was partially supported by the Structured Program of R&D&I INNOVMAR - Innovation and Sustainability in the Management and Exploitation of Marine Resources . The post-doctoral scholarship granted by Erasmus Mundus Interweave to T. de Sousa is greatly acknowledged. C. Magalhães acknowledges Investigator FCT program supported by FCT, funded by the European Social Fund through the Operational Program of Human Capital (POCH) . C. Teixeira acknowledges FCT for a postdoctoral grant (ref. SFRH/BPD/110730/2015 ) cofunded by MCTES and the European Social Fund through POCH/QEC . Appendix A

This work aimed to evaluate the potential of constructed wetlands (CWs) for removal of antibiotics (enrofloxacin and oxytetracycline) and antibiotic resistant bacteria from saline aquaculture wastewaters. Removal of other contaminants (nutrients, organic matter and metals) and toxicity reduction and the influence of antibiotics with these processes were evaluated. Thus, nine CWs microcosms, divided into three treatments, were assembled and used to treat wastewater (doped or not with the selected antibiotics) between October and December of 2015. Each week treated wastewater was removed and new wastewater (doped or not) was introduced in CWs. Results showed > 99% of each antibiotic was removed in CWs. After three weeks of adaptation, removal percentages > 95% were also obtained for total bacteria and for antibiotic resistant bacteria. Nutrients, organic matter and metal removal percentages in CWs treated wastewater were identical in the absence and in the presence of each antibiotic. Toxicity in treated wastewaters was significantly lower than in initial wastewaters, independently of antibiotics presence. Results showed CWs have a high efficiency for removing enrofloxacin or oxytetracycline as well as antibiotic resistant bacteria from saline aquaculture wastewaters. CWs can also remove other contaminants independently of drug presence, making the aquaculture wastewater possible to be reutilized and/or recirculated. © 2016 by the authors. This research was partially supported by the Strategic Funding UID/Multi/04423/2013 through national funds provided by FCT—Foundation for Science and Technology and European Regional Development Fund (ERDF), in the framework of the program PT2020 and by the structured Program of R&D&I INNOVMAR—Innovation and Sustainability in the Management and Exploitation of Marine Resources, reference NORTE-01-0145-FEDER-000035, namely within the Research Lines ECOSERVICES (Assessing the environmental quality, vulnerability and risks for the sustainable management of the NW coast natural resources and ecosystem services in a changing world) and INSEAFOOD (Innovation and Valorisation of Seafood Products: Meeting Local Challenges and Opportunities) within the R&D Institution CIIMAR (Interdisciplinary Centre of Marine and Environmental Research), supported by the Northern Regional Operational Program (NORTE2020), through the European Regional Development Fund (ERDF).