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There is limited research on the occurrence of microplastics (MPs) in canned seafood. All types of canned seafood investigated in the present study were contaminated. After sample digestion in 30 % hydrogen peroxide, a total of 40 MPs were recovered. Fibers were the most common type, blue was the dominant colour, and Fourier Transform Infrared Spectroscopy (FTIR) identified polyester as the most common polymer. Considering all samples, an average of 3.5 ± 5.2 MPs/can was obtained, with octopus in tomato sauce and tuna in olive oil presenting the highest contamination (5.2 ± 7.5 MPs/can and 5.2 ± 5.1 MPs/can, respectively). Also, significant differences between the number of MPs in the seafood tissues and immersion liquids were verified. The present study demonstrates MPs occurrence in canned seafood, a potential contamination pathway for humans. More research on the different stages of the canning processing is vital for understanding MPs contamination in cans., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Microplastics (MPs) are emerging contaminants of increasing concern as they may cause adverse effects and carry other contaminants, which may potentially compromise human health. Despite occurring in aquatic ecosystems worldwide, the knowledge about MP presence in different aquaculture systems and their potential impact on seafood products is still limited. This study aimed to determine the levels of MPs in water, feed, and European seabass (Dicentrarchus labrax) from three relevant aquaculture systems and estimate human exposure to MPs and metals through seabass consumption. The recirculating aquaculture system (RAS) had the highest MP occurrence in water and feed. MP levels in seabass followed the aquaculture system's levels in water and feed, with RAS-farmed fish presenting the highest MP load, both in the fish gastrointestinal tract (GIT) and muscle, followed by pond-, and cage-farmed fish. MPs' characteristics across aquaculture systems and fish samples remained consistent, with the predominant recovered particles falling within the MP size range. The particles were visually characterized and chemically identified by micro-Fourier Transform Infrared Spectroscopy (μFTIR). Most of these particles were fibres composed of man-made cellulose and PET. MP levels in GIT were significantly higher than in muscle for pond- and RAS-farmed fish, MPs' bioconcentration factors >1 indicated bioconcentration in farmed seabass. Metal concentrations in fish muscle were below permissible limits, posing low intake risks for consumers according to the available health-based guidance values and estimated dietary scenarios., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Given the increasing pressure on water bodies, it is imperative to explore sustainable methodologies for wastewater treatment and reuse. The simultaneous presence of multiples contaminants in complex wastewater, such as the liquid effluents from biogas plants, can compromise biological treatment effectiveness for reclaiming water. Vertical subsurface flow constructed wetlands were established as low-cost decentralized wastewater treatment technologies to treat the liquid fraction of digestate from municipal organic waste with metals, antibiotics, and antibiotic resistance genes, to allow its reuse in irrigation. Twelve lab-scale planted constructed wetlands were assembled with gravel, light expanded clay aggregate and sand, testing four different treating conditions (liquid digestate spiked with oxytetracycline, sulfadiazine, or ofloxacin, at 100 μg/ L, or without dosing) during 3 months. Physicochemical parameters (pH, chemical oxygen demand (COD), nutrients, metals, and antibiotics), the microbial communities dynamics (through 16S high-throughput sequencing) and antibiotic resistance genes removal (qPCR) were monitored in influents and effluents. Systems removed 85.8%-96.9% of organic matter (as COD), over 98.1% of ammonium and phosphate ions, and 69.3%-99.4% of nitrate and nitrite ions, with no significant differences between the presence or absence of antibiotics. Removal of Fe, Mn, Zn, Cu, Pb and Cr exceeded 82% in all treatment cycles. The treatment also removed oxytetracycline, sulfadiazine and ofloxacin over 99%, and decreased intl1, tetA, tetW , sul1 and qnrS gene copies. Nonetheless, after 3 months of ofloxacin dosing, qnrS gene started being detected. Removal processes relied on high HRT (14 days) and various mechanisms including sorption, biodegradation, and precipitation. Microbial community diversity in liquid digestate changed significantly after treatment in constructed wetlands with a decrease in the initial Firmicutes dominance, but with no clear effect of antibiotics on the microbial community structure. Removals above 85% and 94% were observed for Streptococcus and Clostridium, respectively. Results suggest that vertical subsurface flow constructed wetlands were a suitable technology for treating the liquid digestate to reuse it in irrigation agricultural systems, contributing to the circular bioeconomy concept. However, a more profound understanding of effective wastewater treatment strategies is needed to avoid antibiotic resistance genes dissemination., Competing Interests: MA and RC were employed by TratoLixo—Tratamento de Resíduos Sólidos, E.I.M. S.A. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Porras-Socias, Tomasino, Fernandes, De Menezes, Fernández, Collins, Alves, Castro, Gomes, Almeida and Mucha.)

Following a fuel leakage inside a Portuguese maritime port, we conducted parallel 30-day experiments using contaminated seawater and fuel, sampled five days after the incident. This study aimed to (i)survey the native microbial community response to the spilled fuel and (ii)evaluate the efficacy of bioremediation, both biostimulation and bioaugmentation with a lyophilized bacterial consortium (Rhodococcus erythropolis, Pseudomonas sp.), in accelerating hydrocarbon degradation. Metabarcoding analysis revealed a shift in microbial communities, with increased abundance of hydrocarbon-degraders (e.g. Alcanivorax, Thalassospira). Ninety-five hydrocarbonoclastic bacteria were isolated, including key groups from the enriched communities. The lyophilized bacteria added in bioaugmentation, enhanced the abundance of hydrocarbon-degraders over time and were recovered throughout time. Bioremediation treatments favoured biodegradation, achieving over 60 % removal of total petroleum hydrocarbons after 15 days, contrasting with natural attenuation where almost no TPH was removed. This work highlights the potential of bioremediation technologies to accelerate hydrocarbon-degrading activity, for oil spills inside ports., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

This comprehensive scientific review evaluates the effectiveness of nature-based solutions (NBS) in reducing antibiotics (ABs), combating antimicrobial resistance (AMR), and controlling pathogens in various aquatic environments at different river catchment levels. It covers conventional and innovative treatment wetland configurations for wastewater treatment to reduce pollutant discharge into the aquatic ecosystems as well as exploring how river restoration and saltmarshes can enhance pollutant removal. Through the analysis of experimental studies and case examples, the review shows NBS's potential for providing sustainable and cost-effective solutions to improve the health of aquatic ecosystems. It also evaluates the use of diagnostic indicators to predict NBS effectiveness in removing specific pollutants such as ABs and AMR. The review concludes that NBS are feasible for addressing the new challenges stemming from human activities such as the presence of ABs, AMR and pathogens, contributing to a better understanding of NBS, highlighting success stories, addressing knowledge gaps, and providing recommendations for future research and implementation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

The antidiabetic drug metformin and antiepileptic drug lamotrigine are contaminants of emerging concern that have been detected in biowaste-derived amendments and in the environment, and their fate must be carefully studied. This work aimed to evaluate their sorption behaviour on soil upon digestate application. Experiments were conducted on soil and digestate-amended soil as a function of time to study kinetic processes, and at equilibrium also regarding the influence of trace metals (Pb, Ni, Cr, Co, Cu, Zn) at ratio pharmaceutical/metal 1/1, 1/10, and 1/100. Pharmaceutical desorption experiments were also conducted to assess their potential mobility to groundwater. Results revealed that digestate amendment increased metformin and lamotrigine adsorbed amounts by 210% and 240%, respectively, increasing organic matter content. Metformin adsorption kinetics were best described by Langmuir model and those of lamotrigine by Elovich and intraparticle diffusion models. Trace metals did not significantly affect the adsorption of metformin in amended soil while significantly decreased that of lamotrigine by 12-39%, with exception for Cu 2+ that increased both pharmaceuticals adsorbed amounts by 5 - 8%. This study highlighted the influence of digestate amendment on pharmaceutical adsorption and fate in soil, which must be considered in the circular economy scenario of waste-to-resource., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Spatial patterns and temporal trends of the butyltin compounds tributyltin (TBT), dibutyltin (DBT), and monobutyltin (MBT) were investigated in a set of sediment samples collected along the SW Portuguese continental shelf. This region did not reach the Good Environmental Status (GES) in accordance with the Marine Strategy Framework Directive (MSFD) during a first evaluation carried out in 2012. Overall, MBT and DBT were the predominant organotin species detected, but high concentrations of TBT were found in and around disposal sites for dredge sludge derived from the dredging in navigation channels, harbours, and shipyard facilities of the Tagus and Sado estuaries. Although Portuguese regulations for monitoring sediment quality in relation to dredging activities consider only PAH, PCB and HCB, they also dictate that other organic contaminants such as butyltin compounds (BTs) should be monitored if suspicion of high values exists, but no action limits are defined for these (MAOTDR, 2007). Without action limits, the monitoring recommendation given in the regulations is not put into practice. Considering their toxicity, BT derivates should be integrated in the legislation, because they represent an environmental threat in the relocation of dredged material, especially when derived from harbour and shipyards areas. Based on this study, we recommend giving more attention to the amounts and impacts of BTs in sediments at dredged material disposal sites (DMDS) and their surroundings. Or even better, in order to be more efficient, monitoring should be done at the source of the dredged materials and not at the sink. In case it is not done, the monitoring of concentrations of TBT (and other BTs) in sediments and organisms, including imposex studies, at all Portuguese sites for disposal of dredged material receiving slightly to strongly contaminated dredged material must be developed., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationship that could have appeared to influence the work reported in this manuscript., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Two of the groups most impacted by microplastics (MPs) are zooplankton and fish larvae, either through MPs ingestion or absorption. Although there has been an increase of studies focusing on MPs ingestion by these organisms, there is still no standardized methodology for the quantification of MPs present in plankton. For example, some reagents normally used to digest plankton and recover MPs appear adversely to affect some plastic characteristics. This can potentially lead to underestimating the amount and types of MPs present in the organisms analyzed. Hence, this work aimed to optimize a methodology to quantify MPs present in plankton, namely zooplankton and fish larvae, and ensuring MPs integrity. Hence, the planktonic organism tissues were digested using 30% (v/v) H 2 O 2 solution at different temperatures and incubation periods while preserving the integrity and polymer characteristics of 13 types of MPs. MPs' characteristics were register before and after the tests, by visual inspection and Fourier Transform Infrared Spectroscopy (FTIR) analysis, to evaluate the integrity and features of MPs. With this methodology, MPs recovery was above 85% for all types of plastic tested. The proposed methodology is a rapid protocol, with a maximum of 7 h of incubation, that ensures simultaneously the full digestion of the organism tissues and the complete preservation of all the plastic characteristics, namely color, size and polymer type.•A methodology was optimized to quantify microplastics present in zooplankton (copepods, chaetognaths and fish larvae).•Thirteen types of microplastics (fibers and fragments of different polymers) were used to test the efficiency of the methodology ensuring the maintenance of the integrity of plastics.•With this methodology, microplastic recovery was above 85% for all the types of microplastic tested and no changes in their characteristics were observed., 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., (© 2023 The Authors. Published by Elsevier B.V.)

The adsorptive performance of mesoporous silica-based materials towards inorganic (metal ions) and organic (dyes) water pollutants was investigated. Mesoporous silica materials with different particle size, surface area and pore volume were prepared and tailored with different functional groups. These materials were then characterised by solid-state techniques, namely vibrational spectroscopy, elemental analysis, scanning electron microscopy and nitrogen adsorption-desorption isotherms, allowing the successful preparation and structural modifications of the materials to be confirmed. The influence of the physicochemical properties of the adsorbents towards the removal of metal ions (Ni 2+ , Cu 2+ and Fe 3+ ) and organic dyes (methylene blue and methyl green) from aqueous solutions was also investigated. The results reveal that the exceptionally high surface area and suitable ζ-potential of the nanosized mesoporous silica nanoparticles (MSNPs) seem to favour the adsorptive capacity of the material for both types of water pollutants. Kinetic studies were performed for the adsorption of organic dyes by MSNPs and large-pore mesoporous silica (LPMS), suggesting that the process follows a pseudo-second-order model. The recyclability along consecutive adsorption cycles and the stability of the adsorbents after use were also investigated, showing that the material can be reused. Current results show the potentialities of novel silica-based material as a suitable adsorbent to remove pollutants from aquatic matrices with an applicability to reduce water pollution.

A bioremediation approach with tide simulation for buried and surface oil degradation was tested for removal of two, three and four rings polycyclic aromatic hydrocarbons (PAHs). Linear models depicted degradation constants of individual PAH as simple additive function of their initial concentrations (C 0 ) in contaminated sand, hydrophobicity, sampling layer and treatment conditions. For all PAHs and treatment conditions, the degradation of oil in buried layers was faster than at the surface. Naturally-occurring microorganisms proved to be efficient for bioremediation of PAHs and were stimulated by fertilizer addition (biostimulation, BS). Bioaugmentation (BA) by addition of a slurry of a native oil-degraders pre-stimulated consortium did not show faster PAH degradation than BS. Degradation was more rapid for PAH present at low C 0 and with intermediate hydrophobicity. Bioremediation of beach sand either with surface or buried crude oil is a cost-effective strategy to clean-up different hydrocarbon families, including persistent ones, such as PAHs., 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.)

Deep-sea mining may lead to the release of high concentrations of metals into the surrounding seabed, which can disturb important ecosystem functions provided by microbial communities. Among these, the production of N2O and its reduction to N2 is of great relevance since N2O is an important greenhouse gas. Metal impacts on net N2O production by deep-sea bacteria are, however, currently unexplored. Here, we evaluated the effects of cadmium (Cd) on net N2O production by a deep-sea isolate, Shewanella loihica PV-4. We performed a series of Cd exposure incubations in oxic conditions and determined N2O fluxes during induced anoxic conditions, as well as the relative expression of the nitrite reductase gene (nirK), preceding N2O production, and N2O reductase gene (nosZ), responsible for N2O reduction. Net N2O production by S. loihica PV-4 exposed to Cd was strongly inhibited when compared to the control treatment (no metal). Both nirK and nosZ gene expression were inhibited in reactors with Cd, but nirK inhibition was stronger, supporting the lower net N2O production observed with Cd. The Cd inhibition of net N2O production observed in this study poses the question whether other deep-sea bacteria would undergo the same effects. Future studies should address this question as well as its applicability to complex communities and other physicochemical conditions, which remain to be evaluated., (© The Author(s) 2023. Published by Oxford University Press on behalf of FEMS.)

Reusing reclaimed wastewater is needed to fight water scarcity, reduce freshwater consumption and conserve water resources, but one must ensure that hazardous substances are fully removed/eliminate before that reuse. The potential of lab-scale constructed wetlands (CWs) for the removal of chemical and biological contaminants from livestock wastewater, while maintaining nutrient levels for fertilization, was assessed, evaluating changes in microbial communities, with particular focus on potential pathogens. CW microcosms with two different substrates (lava rock or light expanded clay aggregate), both planted with Phragmites australis, were tested. After 15 days of treatment, removal rates were higher than 80% for Cd, Cr, Cu, Fe, Pb and Zn, in general with no significant differences between the two different substrates. Organic matter and nutrients were also removed but their levels still allowed the used of the treated wastewater as a fertilizer Removal of bacterial contamination was estimated through enumeration of cultivable bacteria. High removal rates of fecal indicator bacteria were observed, reaching >95% for enterococci and >98% for enterobacteria after 15 days of treatment, decreasing hazardous biological contaminants initially present in the wastewater. In addition, the microbial communities in the initial and treated wastewater, and in the plant roots bed substrate, were characterized by using 16SrRNA gene amplicon sequencing. Microbial communities in the CW systems showed a clear shift comparatively with the initial wastewater showing system adaptation and removal potentialities. This also revealed an important removal of the most represented potential pathogenic genus, Clostridium, which relative abundance decreased from 33% to 1% through the treatment. Overall, CWs showed potential to be efficient in removing chemical and biological contaminants, while maintaining moderated levels of nutrients, allowing the reuse of reclaimed water in agriculture, namely as fertilizer. Current results will contribute for the optimization and use of CWs for a sustainable treatment of liquid wastes, promoting the circular economy., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Fishes are one of the most important components of the oceans and are exposed to several anthropogenic pressures, namely microplastic (MP), contaminants that are now ubiquitous worldwide. Taking advantage of the 2020 Circumnavigation Expedition carried by the NRP Sagres tall ship of the Portuguese Navy, fish samples from the southern Atlantic ocean were collected to evaluate possible MP contamination. In a total of 14 weeks of campaign, seven large migratory fishes of commercial interest were collected across the middle Atlantic Ocean and along the South American Atlantic coast. All individuals were contaminated with MPs, with an average of 18 ± 11 MPs/fish. In all fish sampled, both the gastrointestinal tract and gills presented MPs, indicating different contamination pathways including via their preys and from surrounding water, respectively. A total of 124 MPs were observed, where 72 % were fibers and 28 % particles, mostly of blue color (85 %), and with rayon and nylon as the most abundant polymers. This study is an important contribution to increase the scientific knowledge of MP contamination in mesopelagic fishes used for human consumption and collected in the open waters, reinforcing the need for further research regarding MP contamination in top predatory species from high trophic levels., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Fluorine-based agrochemicals have been benchmarked as the golden standard in pesticide development, prompting their widespread use in agriculture. As a result, fluorinated pesticides can now be found in the environment, entailing serious ecological implications due to their harmfulness and persistence. Microbial degradation might be an option to mitigate these impacts, though environmental microorganisms are not expected to easily cope with these fluoroaromatics due to their recalcitrance. Here, we provide an outlook on the microbial metabolism of fluorinated pesticides by analyzing the degradation pathways and biochemical processes involved, while also highlighting the central role of enzymatic defluorination in their productive metabolism. Finally, the potential contribution of these microbial processes for the dissipation of fluorinated pesticides from the environment is also discussed.

Marine organisms are affected by the ubiquitous occurrence of microplastics (MPs) in the environment. Several protocols have been described to extract and quantify MPs in seafood, although their complex matrices, with high level of fat, can compromise the efficiency of MPs extraction. To solve this issue, the present study aimed to develop a detailed methodology suitable to process seafood samples with different levels of fat, namely fish and molluscs, from fresh and canned sources, including the immersive liquids from the cans. Sample digestion was tested using different solutions (10% KOH, 30% H 2 O 2 ), temperatures (40 °C, 65 °C) and incubation times (24, 48, 72 h). For fat removal, three detergents (two laboratory surfactants and a commercial dish detergent) and 96% ethanol were tested, as well as the manual separation of fat. The methodology optimized in this study combined a digestion with 30% H 2 O 2 at 65 °C, during 24 to 48 h, with a manual separation of the fat remaining after the digestion. All steps from the present methodology were tested in six types of polymers (PE-LD, PET, PE, AC, PS, and lycra), to investigate if these procedures altered the integrity of MPs. Results showed that the optimized methodology will allow for the efficient processing of complex seafood samples with different fat levels, without compromising MPs integrity (recoveries rate higher than 89% for all the polymers tested).

Antibiotics are often applied in aquaculture to prevent fish diseases. These substances can cause disturbances on receiving waters, when not properly eliminated from the aquaculture effluents. In this work, ozone (O 3 ) was investigated as a possible oxidizing agent to remove fishery antibiotics from aquaculture effluents: florfenicol (FF), oxytetracycline (OTC), sulfadimethoxine (SDM), sulfamethoxazole (SMX), and trimethoprim (TMP). Batch experiments were performed using ultrapure water and aquaculture effluents spiked with a mixture of target antibiotics at relatively high concentrations (10 mg L -1 each). OTC, SMX and TMP were fully removed (< 30 min) regardless of the tested conditions, mainly by O 3 direct attack. In contrast, FF was partially removed in 30 min (∼ 10 and 60%, in aquaculture effluents and ultrapure water, respectively), but only in the presence of hydroxyl radicals (HO • ), the FF concentrations reaching levels below the detection limits in ultrapure water after 60 min. In the case of SDM, its degradation was highly influenced by the selected water matrix, but with removals always higher than 68%. In continuous-flow experiments applying more environmentally relevant antibiotic concentrations (100 ng L -1 each) and low O 3 doses (1.5 mg L - 1 ), ozonation highly removed (> 98%) all tested antibiotics from aquaculture effluents with a hydraulic retention time (HRT) of 10 min, except FF (68%). Although by-products were detected in treated samples, zebrafish (Danio rerio) embryotoxicity tests did not show a toxicity increase by applying this ozonation treatment. Ozonation is thus a possible solution to remove antibiotics from aquaculture effluents. Still, full-scale studies in aquaculture farms are needed, and generation of HO • may be favoured to readily oxidize the FF antibiotic., (Copyright © 2022. Published by Elsevier Ltd.)

Aquaculture is the fastest growing animal food-producing sector. Water is the central resource for aquaculture, and it is essential that its quality be preserved. Micropollutants (MPs) can reach aquaculture through anthropogenic addition or inlet water, and may cause harmful effects such as endocrine disruption and antibiotic resistance, adversely affecting the fish species being farmed. Furthermore, the discharge of aquaculture effluents into the environment may contribute to the deterioration of water courses. In this sense, the implementation of environmentally responsible measures in aquaculture farms is imperative for the protection of ecosystems and human health. The European Commission (EC) has recently launched a guiding document promoting ecological aquaculture practices; however, options for water treatment are still lacking. Conventional processes are not designed to deal with MPs; this review article consolidates relevant information on the application of advanced oxidation technologies (AOTs) and constructed wetlands (CWs) as potential strategies in this regard. Although 161 studies on the application of AOTs or CWs in aquaculture have already been published, only 34 focused on MPs (28 on AOTs and 6 on CWs), whereas the others reported the removal of contaminants such as bacteria, organic matter, solids and inorganic ions. No study coupling both treatments has been reported to date for the removal of MPs from aquaculture waters. AOTs and CWs are prospective alternatives for the treatment of aquacultural aqueous matrices. However, the type of aquaculture activity and the specifications of these available technologies should be considered while selecting the most suitable treatment option., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Fluorinated pesticides acquired a significant market share in the agrochemical sector due to the surge of new fluoroorganic ingredients approved in the last two decades. This growing trend has not been accompanied by a comprehensive scientific and regulatory framework entailing all their potential negative impacts for the environment, especially when considering the hazardous properties that may result from the incorporation of fluorine into organic molecules. This review aims to address the safe/hazardous dichotomy associated with fluorinated pesticides by providing an updated outlook on their relevancy in the agrochemical sector and how it leads to their role as environmental pollutants. Specifically, the environmental fate and distribution of these pesticides in the ecosystems is discussed, while also analysing their potential to act as toxic substances for non-target organisms., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Deep-sea sediments (DSS) are one of the largest biotopes on Earth and host a surprisingly diverse microbial community. The harsh conditions of this cold environment lower the rate of natural attenuation, allowing the petroleum pollutants to persist for a long time in deep marine sediments raising problematic environmental concerns. The present work aims to contribute to the study of DSS microbial resources as biotechnological tools for bioremediation of petroleum hydrocarbon polluted environments. Four deep-sea sediment samples were collected in the Mid-Atlantic Ridge, south of the Azores (North Atlantic Ocean). Their autochthonous microbial diversity was investigated by 16S rRNA metabarcoding analysis. In addition, a total of 26 deep-sea bacteria strains with the ability to utilize crude oil as their sole carbon and energy source were isolated from the DSS samples. Eight of them were selected for a novel hydrocarbonoclastic-bacterial consortium and their potential to degrade petroleum hydrocarbons was tested in a bioremediation experiment. Bioaugmentation treatments (with inoculum pre-grown either in sodium acetate or petroleum) showed an increase in degradation of the hydrocarbons comparatively to natural attenuation. Our results provide new insights into deep-ocean oil spill bioremediation by applying DSS hydrocarbon-degrading consortium in lab-scale microcosm to simulate an oil spill in natural seawater.

This work aimed to develop a bioremediation product of lyophilized native bacteria to respond to marine oil spills. Three oil-degrading bacterial strains (two strains of Rhodococcus erythropolis and one Pseudomonas sp.), isolated from the NW Portuguese coast, were selected for lyophilization after biomass growth optimization (tested with alternative carbon sources). Results indicated that the bacterial strains remained viable after the lyophilization process, without losing their biodegradation potential. The biomass/petroleum ratio was optimized, and the bioremediation efficiency of the lyophilized bacterial consortium was tested in microcosms with natural seawater and petroleum. An acceleration of the natural oil degradation process was observed, with an increased abundance of oil-degraders after 24 h, an emulsion of the oil/water layer after 7 days, and an increased removal of total petroleum hydrocarbons (47%) after 15 days. This study provides an insight into the formulation and optimization of lyophilized bacterial agents for application in autochthonous oil bioremediation.

Various contaminants of emerging concern (CECs) have been detected in different ecosystems, posing a threat to living organisms and the environment. Pharmaceuticals are among the many CECs that enter the environment through different pathways, with wastewater treatment plants being the main input of these pollutants. Several technologies for the removal of these pollutants have been developed through the years, but there is still a lack of sustainable technologies suitable for being applied in natural environments. In this regard, solutions based on natural biological processes are attractive for the recovery of contaminated environments. Bioremediation is one of these natural-based solutions and takes advantage of the capacity of microorganisms to degrade different organic pollutants. Degradation of pollutants by native microorganisms is already known to be an important detoxification mechanism that is involved in natural attenuation processes that occur in the environment. Thus, bioremediation technologies based on the selection of natural degrading bacteria seem to be a promising clean-up technology suitable for application in natural environments. In this review, an overview of the occurrence and fate of pharmaceuticals is carried out, in which bioremediation tools are explored for the removal of these pollutants from impacted environments.

Epoxiconazole (EPO) and fludioxonil (FLU) are two widely used fluorinated pesticides known to be highly persistent and with high ecotoxicological potential, turning them into pollutants of concern. This work aimed to optimize two degrading bacterial consortia, previously obtained from an agricultural soil through enrichment with EPO and FLU, by characterizing the contribution of their corresponding bacterial isolates to the biodegradation of these pesticides using both culture-dependent and independent methodologies. Results showed that a co-culture of the strains Hydrogenophaga eletricum 5AE and Methylobacillus sp. 8AE was the most efficient in biodegrading EPO, being able to defluorinate ca. 80% of this pesticide in 28 days. This catabolic performance is likely the result of a commensalistic cooperation, in which H. eletricum may be the defluorinating strain and Methylobacillus sp. may assume an accessory, yet pivotal, catabolic role. Furthermore, 16S rRNA metabarcoding analysis revealed that these strains represent a minority in their original consortium, showing that the biodegradation of EPO can be driven by less abundant phylotypes in the community. On the other hand, none of the tested combinations of bacterial strains showed potential to biodegrade FLU, indicating that the key degrading strains were not successfully isolated from the original enrichment culture. Overall, this work shows, for the first time, the direct involvement of two bacterial species, namely H. eletricum and Methylobacillus sp., in the biodegradation of EPO, while also offering insight on how they might cooperate to accomplish this process. Moreover, the importance of adequate culture-dependent approaches in the engineering of microbial consortia for bioremediation purposes is also emphasized.

Contamination by microplastics is increasing steadily worldwide, affecting all environments. Additionally, aquatic organisms are often exposed to mixtures of other contaminants, including various chemicals. Numerous studies reported adsorption of chemicals to microplastics, raising concern about their possible role as vehicles of exposure through transfer to biota. Nevertheless, until recently, the studies on the topic were mostly focused on the marine environment. In the past five years, however, plenty of publications contributed empirical data about freshwater ecosystems, raising the need for a critical appraisal of the information. Herein the scientific literature was reviewed and multivariate data analysis was done. The analysed studies employed widely different experimental designs, endpoints, test species, shapes and concentrations of various polymer types and chemicals, often not relevant for the freshwater environment. Our integrated analytical approach revealed unfathomable research gaps, given the theoretical knowledge available and lessons learned from research about the marine environment. Greater harmonization of laboratory studies investigating this topic is needed, as well as testing conditions reflecting real exposure scenarios. Furthermore, standardized testing protocols are urgently required to guide such experiments and improve the comparability of the results obtained., (Copyright © 2021 Elsevier B.V. All rights reserved.)

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., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Oil spills are among the most catastrophic events to marine ecosystems and current remediation techniques are not suitable for ecological restoration. Bioremediation approaches can take advantage of the activity of microorganisms with biodegradation capacity thus helping to accelerate the recovery of contaminated environments. The use of native microorganisms can increase the bioremediation efficiency since they have higher potential to survive in the natural environment while preventing unpredictable ecological impacts associated with the introduction of non-native organisms. In order to know the geographical scale to which a native bioremediation consortium can be applied, we need to understand the spatial heterogeneity of the natural microbial communities with potential for hydrocarbon degradation. In the present study, we aim to describe the genetic diversity and the potential of native microbial communities to degrade petroleum hydrocarbons, at an early stage of bioremediation, along the NW Iberian Peninsula coast, an area particularly susceptible to oil spills. Seawater samples collected in 47 sites were exposed to crude oil for 2 weeks, in enrichment experiments. Seawater samples collected in situ , and samples collected after the enrichment with crude oil, were characterized for prokaryotic communities by using 16S rRNA gene amplicon sequencing and predictive functional profiling. Results showed a drastic decrease in richness and diversity of microbial communities after the enrichment with crude oil. Enriched microbial communities were mainly dominated by genera known to degrade hydrocarbons, namely Alcanivorax , Pseudomonas , Acinetobacter , Rhodococcus , Flavobacterium , Oleibacter , Marinobacter , and Thalassospira , without significant differences between geographic areas and locations. Predictive functional profiling of the enriched microbial consortia showed a high potential to degrade the aromatic compounds aminobenzoate, benzoate, chlorocyclohexane, chlorobenzene, ethylbenzene, naphthalene, polycyclic aromatic compounds, styrene, toluene, and xylene. Only a few genera contributed for more than 50% of this genetic potential for aromatic compounds degradation in the enriched communities, namely Alcanivorax , Thalassospira , and Pseudomonas spp. This work is a starting point for the future development of prototype consortia of hydrocarbon-degrading bacteria to mitigate oil spills in the Iberian NW coast., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Bôto, Magalhães, Perdigão, Alexandrino, Fernandes, Bernabeu, Ramos, Carvalho, Semedo, LaRoche, Almeida and Mucha.)

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 >80% for Cd, Cr, Cu, Fe, Mn, and Zn after 2 days of treatment in CWs planted with T. latifolia and >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 >77%, with no significant differences between substrates or plants. Removals of ammonium and phosphate ions ranged between 59-84% and 32-92%, respectively, in CWs with P. australis and 62-75% and 7-68% 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., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

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., 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 © 2019 Elsevier Ltd. All rights reserved.)

This work aimed to investigate the possible effect of 2 quenchers commonly used in H 2 O 2 -based advanced oxidation technologies (AOTs), i.e. catalase and sodium sulphite (Na 2 SO 3 ), 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/H 2 O 2 of 26 micropollutants (MPs) from a spiked surface water (SW), for which the residual H 2 O 2 in the samples was quenched by Na 2 SO 3 , 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 pK a , those monitored after electrospray ionization (ESI) under positive ionization (PI) mode and presenting a pK a higher than 5.9 revealed a great signal suppression, but only when using Na 2 SO 3 as H 2 O 2 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 H 2 O 2 and catalase or Na 2 SO 3 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 pK a higher than 5.9, confirming the interfering effect of H 2 O 2 /Na 2 SO 3 . To the best of our knowledge, the analytical interference in the LC-MS/MS analysis, after adding Na 2 SO 3 to quench H 2 O 2 in AOTs experiments was never reported before and the results presented herein support the recommendation to use catalase instead of Na 2 SO 3 as quencher in AOTs studies., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Paracentrotus lividus gonads are gastronomic delicacies widely appreciated in Europe. Two commercially exploited populations of sea urchins were characterized, for the first time, in terms of gonad quality (market-related traits such as colour, texture and nutritional value) and safety (contaminants levels) to define the best season for harvesting this valuable resource. Spawning occurred between spring-summer, yet gonad index peaked only in one population during winter. In both populations, the strongest yellow colouration and highest gonad firmness, protein, lipid, polyunsaturated fatty acids and carotenoid contents as well as the lowest contents of mineral were registered in fall. Inorganic and organic contaminants levels were, in general, similar among sites and seasons, being found at trace levels in seawater and gonads, with low risk for consumers. For these populations, high-quality gonads can be harvested during fall, despite being safe for human consumption all year round., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

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., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The importance of planktonic microbial communities is well acknowledged, since they are fundamental for several natural processes of aquatic ecosystems. Microorganisms naturally control the flux of nutrients, and also degrade and recycle anthropogenic organic and inorganic contaminants. Nevertheless, climate change effects and/or the runoff of nutrients/pollutants can affect the equilibrium of natural microbial communities influencing the occurrence of microbial pathogens and/or microbial toxin producers, which can compromise ecosystem environmental status. Therefore, improved microbial plankton monitoring is essential to better understand how these communities respond to environmental shifts. The study of marine microbial communities typically involves highly cost and time-consuming sampling procedures, which can limit the frequency of sampling and data availability. In this context, we developed and validated an in situ autonomous biosampler (IS-ABS) able to collect/concentrate in situ planktonic communities of different size fractions (targeting prokaryotes and unicellular eukaryotes) for posterior genomic, metagenomic, and/or transcriptomic analysis at a home laboratory. The IS-ABS field prototype is a small size and compact system able to operate up to 150 m depth. Water is pumped by a micropump (TCS MG2000) through a hydraulic circuit that allows in situ filtration of environmental water in one or more Sterivex filters placed in a filter cartridge. The IS-ABS also includes an application to program sampling definitions, allowing pre-setting configuration of the sampling. The efficiency of the IS-ABS was tested against traditional laboratory filtration standardized protocols. Results showed a good performance in terms of DNA recovery, as well as prokaryotic (16S rDNA) and eukaryotic (18S rDNA) community diversity analysis, using either methodologies. The IS-ABS automates the process of collecting environmental DNA, and is suitable for integration in water observation systems, what will contribute to substantially increase biological surveillances. Also, the use of highly sensitive genomic approaches allows a further study of the diversity and functions of whole or specific microbial communities., Competing Interests: The authors have declared that no competing interests exist.

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., (Copyright © 2019 Elsevier B.V. All rights reserved.)

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 -3 . During the spring-summer period, it was observed the typical increase in the density and diversity of the larval assemblage. Diversity was generally low, with the high dominance of very few taxa, namely the common goby, Pomatoschistus microps. Different types of MPs were found, namely fibers, soft/hard plastic, colorful/transparent plastic, in a total of 2152 particles, with a mean density of 17.06 MPs 100 m -3 . Hard MPs and fibers were the most predominant types, representing 83% of the total MPs collected. In some months the number of MPs surpassed the number of fish larvae, with an average ratio of 1.0 fish larvae:1.5 MPs. Such results are concerning, highlighting that a higher availability of MPs may facilitate their ingestion by fish and therefore increase possible impacts in these communities., (Copyright © 2018 Elsevier B.V. All rights reserved.)

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., (Copyright © 2018 Elsevier B.V. All rights reserved.)

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., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

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., (Copyright © 2018 Elsevier B.V. All rights reserved.)

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., (Copyright © 2018 Elsevier B.V. All rights reserved.)

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., (Copyright © 2018 Elsevier B.V. All rights reserved.)

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., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

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).

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., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

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 (NO 3 - consumption, and NH 4 + 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., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

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., (Copyright © 2017. Published by Elsevier Ltd.)

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., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

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 5months, where the antibiotics were supplemented every three weeks at the concentration of 1mgL -1 , using acetate as a co-substrate. After this period, the acclimated consortia were investigated for their capacity to biodegrade 2 and 3mgL -1 of ENR and CEF. Complete removal of CEF from the inoculated culture medium was always observed within 21days, 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., (Copyright © 2016 Elsevier B.V. All rights reserved.)

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., (Copyright © 2017 Elsevier B.V. All rights reserved.)

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 (0.4m×0.3m×0.3m), simulating CWs, were constructed with Phragmites australis to treat livestock wastewater spiked or not with 100µg/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., (Copyright © 2016 Elsevier Inc. All rights reserved.)

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 (15)N-labeled NO3(-), and the production of (29)N2 and (30)N2 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)., (Copyright © 2016 Elsevier Ltd. All rights reserved.)