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This study aimed to evaluate the potential of a salt marsh plant and its rhizosphere microorganisms for the removal of two pharmaceutical compounds, bezafibrate and paroxetine, from estuarine environment. Plants were exposed for 7 days to a simplified estuarine medium, elutriate solution with or without sediment, doped with bezafibrate or paroxetine. Tests were done in absence and presence of nutrients or copper. Phragmites australis (Cav.) Trin. Ex Steud, alone or with the sediment microbial communities, contributed for pharmaceuticals removal. In the presence of P. australis, for paroxetine a 65% removal was observed. Removal increased up to 90% when sediment was present. For bezafibrate, removals reached ca. 47% in P. australis presence, increasing to ca. 70% when nutrients were added to the medium, indicating a good nutritional state can contribute for a higher compound removal. When Cu was added, 75% removal for bezafibrate and 95% removal for paroxetine were observed indicating the metal might influence the removal of the pharmaceuticals. Overall, the plant and its rhizosediments and associated microorganisms showed potential for pharmaceuticals removal from estuaries, eventually degrading the selected compounds, a feature requiring more research. Results indicate that phytoremediation could be a viable option for eliminating/diminishing the environmental impact of pharmaceutical compounds in estuarine areas. [ABSTRACT FROM AUTHOR]

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

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. [ABSTRACT FROM AUTHOR]

Constructed wetlands (CWs) can be used to reduce various pollutants present in livestock wastewater, such as organic matter, nutrients and metals. Very recently these systems have also been used to remove the so called emergent pollutants. These pollutants can be harmful for both microorganisms and plants, two key players in CWs removal processes. Therefore, the aim of the present work was to assess the influence of emergent pollutants, namely antibiotics, on the removal of pollutants from livestock wastewaters, as antibiotics might decrease CWs performance for the treatment of this type of wastewater. Microcosms (0.4 m × 0.3 m × 0.3 m), simulating CWs, were assembled with Phragmites australis to treat livestock wastewater not doped or doped with 100 μg/L of enrofloxacin and/or of ceftiofur, two antibiotics commonly used in livestock industry. Four different treatments (Control, Enr, Cef and Mix) were tested, each in triplicate. Wastewater was treated during one-week cycle, after which it was removed and replaced by new wastewater (doped or not), in a total of 8 cycles. At weeks 1, 2, 4 and 8 treated wastewater was collected and analysed to determine removal rates of nutrients (ammonia, nitrate, nitrite and phosphate), organic matter (chemical oxygen demand (COD) and biological oxygen demand (BOD)), and solids (including total suspended solids (TSS)), as well as, veterinary antibiotics (enrofloxacin and ceftiofur). High removal rates (up to 90% depending of the parameter) were observed independently of the presence of the veterinary antibiotics, which were also significantly removed from the wastewater. Generally, measured parameters presented values lower than those expressed in the legislation for wastewater discharge into the aquatic environment. Present results indicate that, in tested conditions, the presence of veterinary antibiotics, namely enrofloxacin and ceftiofur, did not influence significantly the biochemical removal processes that occur naturally in CWs during treatment of livestock wastewater, the systems maintaining their performance. Therefore, CWs are a valuable alternative to remove pollutants, including antibiotics, from livestock wastewaters, reducing the impact of this type of effluents into the environment. In addition, this technology can be an efficient/economically viable technology to meet the current wastewater reuse challenges. [ABSTRACT FROM AUTHOR]

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

The increased use of metallic nanoparticles (NPs) raises the probability of finding NPs in the environment. A lot of information exists already regarding interactions between plants and metals, but information regarding interactions between metallic NPs and plants, including salt marsh plants, is still lacking. This work aimed to study interactions between CuO NPs and the salt marsh plants Halimione portulacoides and Phragmites australis . In addition, the potential of these plants for phytoremediation of Cu NPs was evaluated. Plants were exposed for 8 days to sediment elutriate solution doped either with CuO or with ionic Cu. Afterwards, total metal concentrations were determined in plant tissues. Both plants accumulated Cu in their roots, but this accumulation was 4 to 10 times lower when the metal was added in NP form. For P. australis , metal translocation occurred when the metal was added either in ionic or in NP form, but for H. portulacoides no metal translocation was observed when NPs were added to the medium. Therefore, interactions between plants and NPs differ with the plant species. These facts should be taken in consideration when applying these plants for phytoremediation of contaminated sediments in estuaries, as the environmental management of these very important ecological areas can be affected. [ABSTRACT FROM AUTHOR]

Plant roots have the ability to produce and secrete substances, such as aliphatic low molecular weight organic acids (ALMWOAs), into the rhizosphere. This phenomenon occurs for several purposes, for instance, the detoxification of pollutants. Nevertheless, knowledge about the exudation of such substances from marsh plants roots is still scarce. This work aimed at studying: 1) the ability of marsh plants, freshly collected in estuarine marshes, to liberate ALMWOAs into the surrounding medium and 2) the influence of the physiological cycle of these plants on the exudation of those substances. In vitro experiments were carried out, in different seasons, with Phragmites australis and Halimione portulacoides (two marsh plants widely distributed in Europe). Root exudates were collected in freshwater to which plant specimens, in different physiological stages, were exposed. Both marsh plants were capable of liberating oxalic and citric acids into the surrounding medium. Formic acid was also released by P . australis roots and acetic acid by H . portulacoides . There was a seasonal effect on the liberation of ALMWOAs by both plant roots. Marked changes were registered in the nature and levels of the ALMWOAs liberated and such changes depended upon the season in which the specimens were collected. In growing season, a significantly higher liberation of oxalic and citric acids (and acetic acid but only in H . portulacoides case) was observed. For P . australis , formic acid was only found in the decaying stage (autumn and winter). The nature of the medium (in particular, salinity) was a feature conditioning the exudation of ALMWOAs. Both plants were shown to contribute for the presence of ALMWOAs in marsh rhizosediments (some ALMWOAs were found in pore waters extracted). The nature and extent of this contribution will be however dependent upon plants' physiological stage, in addition to plant species. Therefore, these features should be taken into consideration in the event of using marsh plants for phytoremediation purposes in contaminated estuarine areas. [ABSTRACT FROM AUTHOR]

Metals are known to induce oxidative stress in plant cells. Antioxidant thiolic compounds are known to play an important role in plants׳ defence mechanisms against metal toxicity but, regarding salt marsh plants, their role is still very poorly understood. In this work, the involvement of non-protein thiols (NPT), such as cysteine (Cys), reduced glutathione (GSH), oxidised glutathione (GSSG) and total acid-soluble SH compounds (total thiols), in the tolerance mechanisms of the marsh plant Phragmites australis against Cu and Cd toxicity was assessed. Specimens of this plant, freshly harvested in an estuarine salt marsh, were exposed, for 7 days, to rhizosediment soaked with the respective elutriate contaminated with Cu (0, 10 and 100 mg/L) or Cd (0, 1, 10 mg/L). In terms of NPT production, Cu and Cd contamination induced different responses in P. australis . The content of Cys increased in plant tissue after plant exposure to Cu, whereas Cd contamination led to a decrease in GSSG levels. In general, metal contamination did not cause a significant variation on GSH levels. Both metals influenced, to some extent, the production of other thiolic compounds. Despite the accumulation of considerable amounts of Cu and Cd in belowground tissues, no visible toxicity signs were observed. So, antioxidant thiolic compounds were probably involved in the mechanisms used by P. australis to alleviate metal toxicity. As P. australis is considered suitable for phytostabilising metal-contaminated sediments, understanding its tolerance mechanisms to toxic metals is important to optimise the conditions for applying this plant in phytoremediation procedures. [ABSTRACT FROM AUTHOR]

Microbial assisted phytoremediation is a promising, though yet poorly explored, new remediation technique. The aim of this study was to develop autochthonous microbial consortia resistant to cadmium that could enhance phytoremediation of salt-marsh sediments contaminated with this metal. The microbial consortia were selectively enriched from rhizosediments colonized by Juncus maritimus and Phragmites australis. The obtained consortia presented similar microbial abundance but a fairly different community structure, showing that the microbial community was a function of the sediment from which the consortia were enriched. The effect of the bioaugmentation with the developed consortia on cadmium uptake, and the microbial community structure associated to the different sediments were assessed using a microcosm experiment. Our results showed that the addition of the cadmium resistant microbial consortia increased J. maritimus metal phytostabilization capacity. On the other hand, in P. australis, microbial consortia amendment promoted metal phytoextraction. The addition of the consortia did not alter the bacterial structure present in the sediments at the end of the experiments. This study provides new evidences that the development of autochthonous microbial consortia for enhanced phytoremediation of salt-marsh sediments contaminated with cadmium might be a simple, efficient, and environmental friendly remediation procedure. [ABSTRACT FROM AUTHOR]

• 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]

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

Abstract: Degradation of petroleum hydrocarbons in colonized and un-colonized sediments by salt marsh plants Juncus maritimus and Phragmites australis collected in a temperate estuary was investigated during a 5-month greenhouse experiment. The efficiency of two bioremediation treatments namely biostimulation (BS) by the addition of nutrients, and bioaugmentation (BA) by addition of indigenous microorganisms was tested in comparison with hydrocarbon natural attenuation in un-colonized and with rhizoremediation in colonized sediments. Hydrocarbon degrading microorganisms and root biomass were assessed as well as hydrocarbon degradation levels. During the study, hydrocarbon degradation in un-colonized sediments was negligible regardless of treatments. Rhizoremediation proved to be an effective strategy for hydrocarbon removal, yielding high rates in most experiments. However, BS treatments showed a negative effect on the J. maritimus potential for hydrocarbon degradation by decreasing the root system development that lead to lower degradation rates. Although both plants and their associated microorganisms presented a potential for rhizoremediation of petroleum hydrocarbons in contaminated salt marsh sediments, results highlighted that nutrient requirements may be distinct among plant species, which should be accounted for when designing cleanup strategies. [Copyright &y& Elsevier]

Abstract: The potential of Phragmites australis was evaluated for the removal of three veterinary drugs, enrofloxacin (ENR), ceftiofur (CEF) and tetracycline (TET), from aquatic mediums. Results showed that the plant promoted the removal of 94% and 75% of ENR and TET, respectively, from wastewater. Microbial abundance estimation revealed that microorganisms were not a major participant. Occurrence of drugs adsorption to plant roots was observed in small extension. Therefore, main mechanisms occurring were drug removal by plant uptake and/or degradation. Present results demonstrated the potential of P. australis-planted beds to be used for removal of pharmaceuticals from livestock and slaughterhouse industries wastewater. [Copyright &y& Elsevier]

Abstract 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. Highlights • Capacity of microbes from rhizosediment of estuary plants to degrade ENR was assessed. • Enriched microbial cultures were able to degrade ENR though not completely. • Exposure to higher ENR concentrations led to lower biodegradation performances. • Degrading strains predominantly belonged to Proteobacteria and Bacteroidetes phyla. • Assemblage of isolates from enriched cultures also resulted in ENR biodegradation. [ABSTRACT FROM AUTHOR]

Abstract: Several salt marsh plant species have shown to be able to uptake and concentrate metals in their tissues, showing potential for metal phytoremediation. However, studies in controlled conditions, mimicking as much as possible the plants natural environment, are needed to confirm this potential. For the present study, Juncus maritimus and Phragmites australis were collected in an estuary together with the sediment surrounding their roots, put in vessels and maintained in greenhouses under estuarine tidal simulation. After 3 weeks of acclimation, vessels were spiked with two different cadmium concentrations. After 2 months, cadmium was assessed in plant tissues and sediments. Results indicate that both plant species were able to uptake and translocate cadmium into their tissues, contributing also to retain it in rhizosediments and thus reducing the available amount of metal in the environment. Metal was preferentially accumulated in belowground structures, in concentrations not directly proportional to the amount of cadmium present in the sediment. Although no visual toxicity signs were observed, some defence mechanisms were triggered as observed by the changes in carotenoids, lignin, total soluble phenolic compounds and thiolic compounds levels, this response differing between plant species. This work shows that these two salt marsh plants can contribute for the retention of cadmium in salt marshes being useful for the phytostabilization of this metal in estuarine environments. [Copyright &y& Elsevier]

The recovery of estuarine environments is in need. Phytoremediation could be a valid option to reduce pollution while preserving natural biodiversity. In this work, estuarine sediments colonized by Juncus maritimus or Phragmites australis were spiked with cadmium in the absence and in the presence of an autochthonous microbial consortium resistant to the metal. The aim of this study was to increase the potential for cadmium phytoremediation that these two halophyte plants have shown. Experiments were carried out in greenhouses with an automatic irrigation system that simulated estuarine tidal cycles. After 2 months, Cd concentration in P. australis stems increased up to 7 times when the rhizosphere was inoculated with the microbial consortium. So, P. australis phytoextraction potential was increased through autochthonous bioaugmentation. As for J. maritimus, up to 48% more Cd (total amount) was observed in its belowground tissues after being subjected to autochthonous bioaugmentation. Therefore, the phytostabilization potential of this plant was promoted. For both plants this increase in cadmium uptake did not cause significant signs of toxicity. Therefore, the addition of autochthonous microorganisms resistant to cadmium seems to be a valuable strategy to potentiate phytoremediation of this metal in saltmarshes, being useful for the recovery of moderately impacted estuaries. This will contribute for an effective management of these areas. Research on this topic regarding estuarine ecosystems, especially saltmarshes, is, to our knowledge, inexistent. [Copyright &y& Elsevier]

Abstract: The aim of the present work was to evaluate, at microcosm level, the capacity of constructed wetlands (CWs) to remove veterinary pharmaceutical compounds, from wastewater. Results indicated that CWs have potential to mitigate the release of veterinary drugs, namely enrofloxacin (ENR, a fluoroquinolone) and tetracycline (TET, tetracyclines family). Removal efficiencies of 94% and 98% where achieved for TET and ENR, respectively, when treating pigfarm wastewater effluent doped at 100μgL−1 drug level, along twelve weeks. Occurrence of adsorption of the drugs to CWs substrate may be the predominant mechanism for ENR, although for TET there are signs that degradation is also occurring. [Copyright &y& Elsevier]