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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.)

Potential effects of metal nanoparticles on aquatic organisms and food webs are hard to predict from the results of single-species tests under controlled laboratory conditions, and more realistic exposure experiments are rarely conducted. We tested whether silver nanoparticles (Ag NPs) had an impact on zooplankton grazing on their prey, specifically phytoplankton and bacterioplankton populations. If Ag NPs directly reduced the abundance of prey, thereby causing the overall rate of grazing by their predators to decrease, a cascading effect on a planktonic estuarine food web would be seen. Our results show that the growth rates of both phytoplankton and bacterioplankton populations were significantly reduced by Ag NPs at concentrations of ≥500 μg L(-1). At the same time, grazing rates on these populations tended to decline with exposure to Ag NPs. Therefore, Ag NPs did not cause a cascade of effects through the food web but impacted a specific trophic level. Photosynthetic efficiency of the phytoplankton was significantly reduced at Ag NPs concentrations of ≥500 μg L(-1). These effects did not occur at relatively low concentrations of Ag that are often toxic to single species of bacteria and other organisms, suggesting that the impacts of Ag NP exposure may not be apparent at environmentally relevant concentrations due to compensatory processes at the community level.

The fate of excess nitrogen in estuaries is determined by the microbial-driven nitrogen cycle, being denitrification a key process since it definitely removes fixed nitrogen as N2. However, estuaries receive and retain metals, which may negatively affect this process efficiency. In this study, we evaluated the role of salt marsh plants in mediating cadmium (Cd) impact on microbial denitrification process. Juncus maritimus and Phragmites australis from an estuary were collected together with the sediment involving their roots, each placed in vessels and maintained in a greenhouse, exposed to natural light, with tides simulation. Similar non-vegetated sediment vessels were prepared. After 3 weeks of accommodation, nine vessels (three per plant species plus three non-vegetated) were doped with 20 mg/L Cd(2+) saline solution, nine vessels were doped with 2 mg/L Cd(2+) saline solution and nine vessels were left undoped. After 10 weeks, vessels were dissembled and denitrification potential was measured in sediment slurries. Results revealed that the addition of Cd did not cause an effect on the denitrification process in non-vegetated sediment but had a clear stimulation in colonized ones (39 % for P. australis and 36 % for J. maritimus). In addition, this increase on denitrification rates was followed by a decrease on N2O emissions and on N2O/N2 ratios in both J. maritimus and P. australis sediments, increasing the efficiency of the N2O step of denitrification pathway. Therefore, our results suggested that the presence of salt marsh plants functioned as key mediators on the degree of Cd impact on microbial denitrification.

The present work aims to evaluate the MP contamination of zooplankton and its impact on MP trophic transfers at the lower levels of the food web in a field study. During 1 year, seasonal surveys were conducted to collect zooplankton and water samples from different sites in two estuaries, the Douro and Lima estuaries (NW, Portugal). The zooplankton was quantified and identified into major zooplanktonic groups. Dedicated protocols that had been previously optimized were used to assess the MP presence in the water samples and in two of the most abundant zooplankton groups (copepods and chaetognaths). The results showed the presence of MPs in all water samples, with similar MP concentrations in both estuaries (Lima: 2.4 ± 2.0 MPs m−3; Douro: 2.3 ± 1.9 MPs m−3). In general, no temporal or spatial variations were observed. Fibres, blue and of a small size (<1 mm), were the most common characteristics of the MPs found in the water and zooplankton, indicating that water can be a source of MPs for zooplankton. Chaetognatha exhibited higher MP contamination in the Lima (2.9 ± 3.1 MPs ind−1) and Douro (2.0 ± 2.8 MPs ind−1) estuaries than Copepoda, which tended to have lower levels of MP contamination (Lima: 0.95 ± 1.12 MPs ind−1; Douro: 1.1 ± 1.2 MPs ind−1). Such differences in the MP concentrations between these two categories of zooplanktonic organisms indicate a possible MP trophic transfer at the lower levels of the food web. The results highlight the novel possibility of an MP trophic transfer in zooplankton and the need to fully assess the impacts of MPs in real scenarios. [ABSTRACT FROM AUTHOR]

Concerns regarding plastic pollution, especially microplastics, have increased, as they can be present in different environmental compartments, including estuarine areas and saltmarshes. Although saltmarshes are highly vulnerable to different human activities and pressures, they have the ability to trap/retain contaminants in their vegetated sediments. However, there is still little information regarding the role of saltmarshes in microplastic retention. Thus, the present study aims to investigate the capability of an estuarine saltmarsh to trap microplastics by comparing microplastic concentrations in vegetated (saltmarsh) and non-vegetated sediments. Microplastic content from sediment (vegetated and non-vegetated) samples collected at different sampling sites in Lima River estuary was estimated using previously optimised extraction protocols, and the observed particles were then characterised accordingly to their size, colour, shape, and polymer (by FTIR). Water samples were also collected and analysed for their microplastics content to complement MPs characterisation within the estuarine area. Microplastics were detected in all sediment samples, with fibres being the most common type of microplastic found, followed by fragments/particles. Overall, vegetated sediments, especially those of saltmarsh species Juncus maritimus, presented a higher number of plastic items. These results indicated that microplastics tend to be trapped in vegetated sediments, supporting the fact that saltmarshes have a significant influence on the transport, distribution, and accumulation of MPs in estuarine areas. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Salt marsh plants and associated microorganisms can have an important role in contaminant removal from estuaries, through bioremediation processes. Nevertheless, the interaction between emerging contaminants, namely antibiotics, and plant–microorganism associations in estuarine environment are still scarcely known. In this vein, the aim of the present study was to evaluate, in controlled conditions, the response of a salt marsh plant–microorganism association to a contamination with a veterinary antibiotic. For that a salt marsh plant ( Phragmites australis ) and its respective rhizosediment were collected in a temperate estuary (Lima estuary, NW Portugal) and exposed for 7 days to enrofloxacin (ENR) under different nutritional conditions in sediment elutriates. Response was evaluated in terms of ENR removal and changes in microbial community structure (evaluated by ARISA) and abundance (estimated by DAPI). In general, no significant changes were observed in microbial abundance. Changes in bacterial richness and diversity were observed but only in unplanted systems. However, multivariate analysis of ARISA profiles showed significant effect of both the presence of plant and type of treatment on the microbial community structure, with significant differences among all treatment groups. In addition, plants and associated microorganisms presented a potential for antibiotic removal that, although highly dependent on their nutritional status, can be a valuable asset to recover impacted areas such as estuarine ones. [ABSTRACT FROM AUTHOR]

Estuaries are often considered sinks for contaminants and the cleanup of salt marshes, sensitive ecosystems with a major ecological role, should be carried out by means of least intrusive approaches, such as bioremediation. This study was designed to evaluate the influence of plant-microorganisms associations on petroleum hydrocarbons fate in salt marshes of a temperate estuary (Lima River, NW Portugal). Sediments un-colonized and colonized (rhizosediments) by different plants (Juncus maritimus, Phragmites australis, Triglochin striata and Spartina patens) were sampled in four sites of the lower and middle estuary for hydrocarbon degrading microorganisms (HD), total cell counts (TCC) and total petroleum hydrocarbons (TPHs) assessment. In general, TPHs, HD and TCC were significantly higher ( P < 0.05) in rhizosediments than in un-colonized sediments. Also recorded were differences on the abundance of hydrocarbon degraders among the rhizosediment of the different plants collected at the same site ( J. maritimus < P. australis < T. striata), with statistically significant differences ( P < 0.05) between J. maritimus and T. striata. Moreover, strong positive correlations-0.81 and 0.84 ( P < 0.05), between biotic (HD) and abiotic (organic matter content) parameters and TPHs concentrations were also found. Our data clearly suggest that salt marsh plants can influence the microbial community, by fostering the development of hydrocarbon-degrading microbial populations in its rhizosphere, an effect observed for all plants. This effect, combined with the plant capability to retain hydrocarbons around the roots, points out that salt marsh plant-microorganisms associations may actively contribute to hydrocarbon removal and degradation in estuarine environments. [ABSTRACT FROM AUTHOR]

Abstract: This work aimed to evaluate, in vitro, the capability of roots of two salt marsh plants to release low molecular weight organic acids (LMWOAs) and to ascertain whether Cu contamination would stimulate or not organic acids exudation. The sea rush Juncus maritimus and the sea-club rush Scirpus maritimus, both from the lower Douro river estuary (NW Portugal), were used. Plants were collected seasonally, four times a year in 2004, during low tide. After sampling, plant roots were washed for removal of adherent particles and immersed for 2 h in a solution that matched salinity (3) and pH (7.5) of the pore water from the same location to obtain plant exudates. In one of the seasons, similar experiments were carried out but spiking the solution with different amounts of Cu in order to embrace the range between 0 and 1600 nM. In the final solutions as well as in sediment pore water LMWOAs were determined by high performance liquid chromatography. Plants were able to release, in a short period of time, relatively high amounts of LMWOAs (oxalate, citrate, malate, malonate, and succinate). In the sediment pore water oxalate, succinate and acetate were also detected. Therefore, plant roots probably contributed to the presence of some of these organic compounds in pore water. Exudation differed between the plant species and also showed some seasonally variation, particularly for S. maritimus. The release of oxalate by J. maritimus increased with Cu increase in the media. However, exudation of the other LMWOAs did not seem to be stimulated by Cu contamination in the media. This fact is compatible with the existence of alternative internal mechanisms for Cu detoxification, as denoted by the fact that in media contaminated with Cu both plant species accumulated relatively high amounts (29–83%) of the initially dissolved Cu. This study expands our knowledge on the contribution of globally dominant salt marsh plants to the release of LMWOAs into the environment. [Copyright &y& Elsevier]

Abstract: The influence of each of four organic pollutants selected from among those commonly found in coastal areas, 1,1-dichloro-2,2-bis(p-chlorophenyl) ethylene (DDE), monobutyltin (MBT), Triton X-100 and polycyclic aromatic hydrocarbons (PAHs), on Cu accumulation by Halimione portulacoides was investigated. Experiments were carried out in a laboratory setting, either in hydroponics (sediment elutriate) or in a salt marsh sediment (Cávado River, NW Portugal) soaked in elutriate. Groups of H. portulacoides were exposed to media for 6 days spiked with 10mg/L Cu(II) and with one of the selected pollutants, at an environmentally realistic concentration. DDE and MBT did not cause any major change on Cu accumulation by H. portulacoides, whereas PAHs slightly increased accumulation only in hydroponics i.e. in the absence of sediment. On the other hand, the non-ionic surfactant Triton X-100 markedly favoured Cu accumulation on plant roots both in the presence and absence of sediment. The addition of DDE, MBT and Triton X-100 also favoured Cu solubility from sediments. Therefore, the simultaneous presence of pollutants from different nature (inorganic and organic) in the estuarine environment may result in a composition of water column, pore water, sediment or biota different of that expected considering the effect of each individual pollutant. [Copyright &y& Elsevier]

Total-recoverable metals (Al, Cr, Cu, Fe, Mn, Ni, Pb and Zn) in sediments from Minho estuary salt marsh were determined to evaluate possible increase in anthropogenic contamination by metals and to evaluate the possibility of this area to be considered a pristine area in terms of metals, which can be used as a reference site for other metal-contaminated national and international estuaries/salt marshes. This study revealed that the spatial distribution of metals in the salt marsh sediments was not homogeneous and that two sampling sites (sites 5 and 7) had indications of anthropogenic contamination. However, metal levels in these salt marsh sediments were lower than those observed in the wetlands of the main Portuguese estuaries. Comparison with Portuguese and international reference values used in the evaluation of the ecological quality of sediments, indicated that the sediments can be classified as “clean sediment” and that metal levels were lower or similar (only for Cu and Ni) to the values of ERL, which are the values that define the concentrations ranges that are rarely associated to adverse biologic effects in organisms. In addition, metal levels in the sediments were in chemical forms that were not easily available to organisms, indicating that these sediments probably will not have negative influences in the organisms living in the salt marsh, although no life-form ecological safety tests have been carried out. Therefore, the Minho estuary salt marsh area can probably be considered a pristine area in terms of metals and can be used as a reference for other metal-contaminated estuaries/salt marshes. [ABSTRACT FROM AUTHOR]

Influence of Halimione portulacoides, commonly found in temperate salt marshes, on sediment metal contents, speciation and potential mobility in case of sediment re-suspension was evaluated. Both colonized and non-colonized sediments were studied for total Cd, Cu, Pb and Zn contents and metal fraction exchangeable to water collected in situ. Sediment elutriates, prepared with water collected from each site, were used to simulate a sediment re-suspension phenomenon. As the characteristics and degree of contamination of sediments may influence system behaviour, salt marshes of two Portuguese estuaries, Cavado (NW coast) and Sado (SW coast), were studied. Cu, Pb and Zn contents higher than ERL (quality guideline, effect range-low) were observed, indicating potential risks for living organisms. Strong Cu-complexing organic ligands, also determined in both water and elutriates, were higher in rhizosediment elutriates, at concentrations similar, or even higher, to those of Cu. Such ligands condition metals speciation in the water column and probably also metal bioavailability. From rhizosediment significant amounts of Cu and Zn were transferred to the aqueous phase, concentrations 2–8 times higher than concentrations present in water. In contrast, elutriates of non-colonized sediment removed metals from water, Cu and Zn levels in elutriates being 2–6 times lower than initial ones. Cd and Pb levels in water and elutriates were not measurable in most cases. Results clearly indicate that metals potential solubility in the rhizosphere of plants was markedly higher than that in the surrounding sediment. The obtained results indicated that H. portulacoides presence (and probably other salt marsh plants) may cause a marked increase in metals concentrations in dissolved phase (pore water or even water column if rhizosediment is re-suspended). As salt marsh plants may be abundant in temperate and subtropical estuaries and costal lagoons, this phenomenon should not be disregard in future studies towards the sustainable management of such environments. [Copyright &y& Elsevier]

• Effect of polyethylene microspheres (mPE) on metal phytoremediation investigated. • P. australis accumulated identical amounts of Cu and Cd with or without PE MPs. • Polyethylene microbeads did not affect the plant capacity to phytoremediate Cu or Cd. The negative impacts of microplastics (MPs) in aquatic organisms have been stressed out, but investigation on MPs impacts in plants is scarce. This study aimed to investigate possible interferences of polyethylene microbeads (mPE) (intentionally produced MPs) in the phytoremediation of metals. For that, the plant Phragmites australis , a salt marsh plant with known capability for metals phytoremediation, was exposed for seven days to media contaminated with Cu or Cd and mPE. Elutriate solution, with or without sediment, was used to simulate interactions between estuarine water, plants and sediments. Metal contents on plants tissues and media were measured afterwards. Results showed that plants accumulated significant amounts (up to 70 times more than in non-exposed plants) of both metals in their belowground tissues (reaching ca. 1 mg/g of Cu and ca. 70 μg/g of Cd in roots). Metal accumulation occurred either in absence or in presence of sediments, denoting plants capability to phytoremediate Cu and Cd. No significant metal translocation to the upper ground tissues was observed, metals levels being identical to those of non-exposed plants. The accumulation of both metals in plants belowground tissues was identical in the presence and absence of mPE, indicating that mPE did not interfered with plants capacity to phytoremediate Cu or Cd contaminated medium. This study further our scientific understanding of MPs impacts in saltmarsh plants and on their phytoremediation potential. [ABSTRACT FROM AUTHOR]

Abstract: A capillary electrophoretic method for the determination of the amino acid β-N-methylamino-l-alanine (BMAA) was achieved using a fused-silica capillary column (50 cm × 75 μm I.D.) filled with 5 mM sodium tetraborate solution (pH 9), with an applied voltage of 25 kV, at 25 °C. The method was then applied in quantifying BMAA in eighteen strains of lyophilized estuarine cyanobacteria, following amino acid extraction using 0.1 M trichloroacetic acid and 6 M hydrochloric acid, sequentially. [Copyright &y& Elsevier]