Boosting pharmaceutical removal through aeration in constructed wetlands.

This work evaluated the removal efficiency of 13 wastewater-borne pharmaceuticals in a pilot constructed wetland (CW) operated under different aeration strategies (no aeration, intermittent and continuous). Aeration improved the removal of conventional wastewater parameters and the targeted micropollutants, compared to the non-aerated treatment. Reduction of chemical oxygen demand (COD) and total nitrogen (TN) was slightly higher applying intermittent aeration than applying continuous aeration, the opposite was observed for the investigated pharmaceuticals. Seven targeted compounds were found in influent wastewater, and five of them (acetaminophen, diclofenac, ketoprofen, bezafibrate and gemfibrozil) were efficiently removed (> 83%) in the aerated systems. The overall risk of the investigated samples against aquatic ecosystems was moderate, decreasing in the order influent > no aeration > intermittent aeration > continuous aeration, based on the hazard quotient approach. Lorazepam, diclofenac and ketoprofen were the pharmaceuticals that could contribute the most to this potential environmental impact of the CW effluents after discharge. To the authors' knowledge this is the first sound study on the removal and fate of ketoprofen, bezafibrate, and lorazepam in aerated CWs, and provides additional evidence on the removal and fate of acetaminophen, diclofenac, gemfibrozil, and carbamazepine in this type of bioremediation systems at pilot plant scale.

Constructed wetlands operated as bioelectrochemical systems for the removal of organic micropollutants.

The removal of organic micropollutants (OMPs) has been investigated in constructed wetlands (CWs) operated as bioelectrochemical systems (BES). The operation of CWs as BES (CW-BES), either in the form of microbial fuel cells (MFC) or microbial electrolysis cells (MEC), has only been investigated in recent years. The presented experiment used CW meso-scale systems applying a realistic horizontal flow regime and continuous feeding of real urban wastewater spiked with four OMPs (pharmaceuticals), namely carbamazepine (CBZ), diclofenac (DCF), ibuprofen (IBU) and naproxen (NPX). The study evaluated the removal efficiency of conventional CW systems (CW-control) as well as CW systems operated as closed-circuit MFCs (CW-MFCs) and MECs (CW-MECs). Although a few positive trends were identified for the CW-BES compared to the CW-control (higher average CBZ, DCF and NPX removal by 10-17% in CW-MEC and 5% in CW-MFC), these proved to be not statistically significantly different. Mesoscale experiments with real wastewater could thus not confirm earlier positive effects of CW-BES found under strictly controlled laboratory conditions with synthetic wastewaters.

Microalgae-based bioremediation of water contaminated by pesticides in peri-urban agricultural areas.

The present study evaluated the capacity of a semi-closed, tubular horizontal photobioreactor (PBR) to remove pesticides from agricultural run-off. The study was carried out in summer (July) to study its efficiency under the best conditions (highest solar irradiation). A total of 51 pesticides, including 10 transformation products, were selected and investigated based on their consumption rate and environmental relevance. Sixteen of them were detected in the agricultural run-off, and the estimated removal efficiencies ranged from negative values, obtained for 3 compounds, namely terbutryn, diuron and imidacloprid, to 100%, achieved for 10 compounds. The acidic herbicide MCPA was removed by 88% in average, and the insecticides 2,4-D and diazinon showed variable removals, between 100% and negative values. The environmental risk associated to the compounds still present in the effluent of the PBR was evaluated using hazard quotients (HQs), calculated using the average and highest measured concentrations of the compounds. HQ values > 10 (meaning high risk) were obtained for imidacloprid (21), between 1 and 10 (meaning moderate risk) for 2,4-D (2.8), diazinon (4.6) and terbutryn (1.5), and <1 (meaning low risk) for the remaining compounds diuron, linuron and MCPA. The PBR treatment yielded variable removals depending on the compound, similarly to conventional wastewater treatment plants. This study provides new data on the capacity of microalgae-based treatment systems to eliminate a wide range of priority pesticides under real/environmental conditions.

A review of emerging organic contaminants (EOCs), antibiotic resistant bacteria (ARB), and antibiotic resistance genes (ARGs) in the environment: Increasing removal with wetlands and reducing environmental impacts.

Emerging organic contaminants (EOCs) include a diverse group of chemical compounds, such as pharmaceuticals and personal care products (PPCPs), pesticides, hormones, surfactants, flame retardants and plasticizers. Many of these compounds are not significantly removed in conventional wastewater treatment plants and are discharged to the environment, presenting an increasing threat to both humans and natural ecosystems. Recently, antibiotics have received considerable attention due to growing microbial antibiotic-resistance in the environment. Constructed wetlands (CWs) have proven effective in removing many EOCs, including different antibiotics, before discharge of treated wastewater into the environment. Wastewater treatment systems that couple conventional treatment plants with constructed and natural wetlands offer a strategy to remove EOCs and reduce antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) far more efficiently than conventional treatment alone. This review presents as overview of the current knowledge on the efficiency of different wetland systems in reducing EOCs and antibiotic resistance.

Fate of priority pharmaceuticals and their main metabolites and transformation products in microalgae-based wastewater treatment systems.

The present study evaluates the removal capacity of two high rate algae ponds (HRAPs) to eliminate 12 pharmaceuticals (PhACs) and 26 of their corresponding main metabolites and transformation products. The efficiency of these ponds, operating with and without primary treatment, was compared in order to study their capacity under the best performance conditions (highest solar irradiance). Concentrations of all the target compounds were determined in both water and biomass samples. Removal rates ranged from moderate (40-60 %) to high (>60 %) for most of them, with the exception of the psychiatric drugs carbamazepine, the ß-blocking agent metoprolol and its metabolite, metoprolol acid. O-desmethylvenlafaxine, despite its very low biodegradability in conventional wastewater treatment plants, was removed to certain extent (13-39 %). Biomass concentrations suggested that bioadsorption/bioaccumulation to microalgae biomass was decisive regarding the elimination of non-biodegradable compounds such as venlafaxine and its main metabolites. HRAP treatment with and without primary treatment did not yield significant differences in terms of PhACs removal efficiency. The implementation of HRAPs as secondary treatment is a feasible alternative to CAS in terms of overall wastewater treatment, including organic micropollutants, with generally higher removal performances and implying a green, low-cost and more sustainable technology.

Can high rate algal ponds be used as post-treatment of UASB reactors to remove micropollutants?

The present study evaluated the removal capacity of a UASB-HRAP treatment system, combining anaerobic and microalgae-based, aerobic treatment, for eleven organic micropollutants present in raw sewage, including pharmaceuticals, estrogens and xenoestrogens. The UASB reactor and the HRAP were operated at a hydraulic retention time (HRT) of 7 h and 8 days, respectively. Influent and effluent samples from the UASB and HRAP were collected periodically. All the target compounds were detected in raw sewage, with an occurrence ranging from 70 to 100%. Removal rates in the UASB reactor were generally incomplete, ranging from no removal (-25.12% for the hormone EE2-ethinylestradiol) to 84.91% (E2 - estradiol). However, the overall performance of the UASB + HRAP system was highly efficient for the majority of the compounds, with removal rates ranging from 64.8% (ibuprofen) to 95% (estrone). Gemfibrozil and bisphenol A were the only exceptions, with overall removal rates of 39% and 43%, respectively. Hormones were the compounds with the highest removal rates in the system.

Polyhydroxybutyrate and glycogen production in photobioreactors inoculated with wastewater borne cyanobacteria monocultures.

The aim of this study was to investigate the PHB and glycogen accumulation dynamics in two photobioreactors inoculated with different monocultures of wastewater-borne cyanobacteria, using a three-stage feeding strategy (growth phase, feast-famine phase and feast phase). Two cyanobacterial monocultures containing members of Synechocystis sp. or Synechococcus sp. were collected from treated wastewater and inoculated in lab-scale photobioreactors to evaluate the PHB and glycogen accumulation. A third photobioreactor with a complex microbial community grown in real wastewater was also set up. During each experimental phase different concentrations of inorganic carbon were applied to the cultures, these shifts allowed to discern the accumulation mechanism of carbon storage polymers (PHB and glycogen) in cyanobacteria. Conversion of one into the other was directly related to the carbon content. The highest PHB and glycogen contents (5.04%dcw and 69%dcw, respectively) were achieved for Synechocystis sp.

Start-up of a microalgae-based treatment system within the biorefinery concept: from wastewater to bioproducts.

Within the European project INCOVER, an experimental microalgae-based treatment system has been built for wastewater reuse and added-value products generation. This article describes this new experimental plant and the start-up stage, starting from the new design of three semi-closed horizontal photobioreactors with low energy requirements for microalgae cultivation (30 m3 total), using agricultural runoff and urban wastewater as feedstock. The inflow nutrients concentration is adjusted to select cyanobacteria, microalgae able to accumulate polyhydroxybutyrates, which can be used for bioplastics production. Part of the harvested biomass is used as substrate for anaerobic co-digestion (AcoD) with secondary sludge to obtain biogas. This biogas is then cleaned in an absorption column to reach methane concentration up to 99%. The digestate from the AcoD is further processed in sludge wetlands for stabilization and biofertilizer production. On the other hand, treated water undergoes ultrafiltration and disinfection through a solar-driven process, then it is pumped through absorption materials to recover nutrients, and eventually applied in an agricultural field to grow energy crops by means of a smart irrigation system. This plant presents a sustainable approach for wastewater management, which can be seen as a resource recovery process more than a waste treatment.

Production of polyhydroxybutyrates and carbohydrates in a mixed cyanobacterial culture: Effect of nutrients limitation and photoperiods.

In the present study, different photoperiods and nutritional conditions were applied to a mixed wastewater-borne cyanobacterial culture in order to enhance the intracellular accumulation of polyhydroxybutyrates (PHBs) and carbohydrates. Two different experimental set-ups were used. In the first, the culture was permanently exposed to illumination, while in the second it was submitted to light/dark alternation (12 h cycles). In both cases, two different nutritional regimes were also evaluated, N-limitation and P-limitation. Results showed that the highest PHB concentration (104 mg L-1) was achieved under P limited conditions and permanent illumination, whereas the highest carbohydrate concentration (838 mg L-1) was obtained under N limited condition and light/dark alternation. With regard to bioplastics and biofuel generation, this study demonstrates that the accumulation of PHBs (bioplastics) and carbohydrates (potential biofuel substrate) is favored in wastewater-borne cyanobacteria under conditions where nutrients are limited.

Evaluation of the influence of surfactants in the bioaccumulation kinetics of sulfamethoxazole and oxazepam in benthic invertebrates.

The potential ecotoxicological effects of mixtures of contaminants in the aquatic environment are generating a global concern. Benthic invertebrates, such as the crustacean Gammarus fossarum, are key in the functioning of aquatic ecosystems, and are frequently used as sentinel species of water quality status. The aim of this work was to study the effects of a mixture of the most frequently detected surfactants in the bioconcentration kinetics of two pharmaceuticals in G. fossarum, evaluating their potential enhancing or suppressing effects. Laboratory exposure experiments for both pharmaceuticals and surfactants (concentration ratio 1:25) were set up for two individual compounds, the anxiolytic oxazepam and the antibiotic sulfamethoxazole. Gammarid samples were processed using microQuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) extraction. Pharmaceuticals concentration in the organisms was followed-up by means of nanoliquid chromatography coupled to tandem mass spectrometry (nanoLC-MS/MS). Results indicated a similar mode of action of the surfactants in the bioconcentration kinetics of both drugs, decreasing the accumulation rate in the organism. Oxazepam showed a higher accumulation potential than sulfamethoxazole in all cases. Depuration experiments for oxazepam also demonstrated the high depurative capacity of gammarids, eliminating >50% of the concentration of oxazepam in <6h.

Cultivation and selection of cyanobacteria in a closed photobioreactor used for secondary effluent and digestate treatment.

The main objective of this study was to select and grow wastewater-borne cyanobacteria in a closed photobioreactor (PBR) inoculated with a mixed consortium of microalgae. The 30L PBR was fed with a mixture of urban secondary effluent and digestate, and operated in semi-continuous mode. Based on the nutrients variation of the influent, three different periods were distinguished during one year of operation. Results showed that total inorganic nitrogen (TIN), inorganic phosphorus concentration (PO43-), phosphorus volumetric load (LV-P) and carbon limited/non-limited conditions leaded to different species composition, nutrients removal and biomass production in the culture. High TIN/PO43- concentrations in the influent (36mg N L-1/3mg P L-1), carbon limitation and an average LV-P of 0.35mg P L-1d-1 were negatively related to cyanobacteria dominance and nutrients removal. On the contrary, cyanobacteria predominance over green algae and the highest microbial biomass production (averaging 0.084g Volatile Suspended Solids (VSS) L-1d-1) were reached under TIN/PO43- concentrations of 21mg N L-1/2mg P L-1, no carbon limitation and an average LV-P of 0.23mg P-PO43- L-1d-1. However, although cyanobacteria predominance was also favored with a LV-P 0.15mg L-1d-1, biomass production was negatively affected due to a P limitation in the culture, resulting in a biomass production of 0.0.39g VSS L-1d-1. This study shows that the dominance of cyanobacteria in a microalgal cyanobacterial community in an agitated PBR using wastewater as nutrient source can be obtained and maintained for 234days. These data can also be applied in future biotechnology applications to optimize and enhance the production of added value products by cyanobacteria in wastewater treatment systems.

Multiresidue trace analysis of pharmaceuticals, their human metabolites and transformation products by fully automated on-line solid-phase extraction-liquid chromatography-tandem mass spectrometry.

A novel, fully automated analytical methodology based on dual column liquid chromatography coupled to tandem mass spectrometry (LC-LC-MS(2)) has been developed and validated for the analysis of 12 pharmaceuticals and 20 metabolites and transformation products in different types of water (influent and effluent wastewaters and surface water). Two LC columns were used - one for pre-concentration of the sample and the second for separation and analysis - so that water samples were injected directly in the chromatographic system. Besides the many advantages of the methodology, such as minimization of the sample volume required and its manipulation, both compounds ionized in positive and negative mode could be analyzed simultaneously without compromising the sensitivity. A comparative study of different mobile phases, gradients and LC pre-concentration columns was carried out to obtain the best analytical performance. Limits of detection (MLODs) achieved were in the low ngL(-1) range for all the compounds. The method was successfully applied to study the presence of the target analytes in different wastewater and surface water samples collected near the city of Girona (Catalonia, Spain). Data on the environmental presence and fate of pharmaceutical metabolites and TPs is still scarce, highlighting the relevance of the developed methodology.

Occurrence and in-stream attenuation of wastewater-derived pharmaceuticals in Iberian rivers.

A multitude of pharmaceuticals enter surface waters via discharges of wastewater treatment plants (WWTPs), and many raise environmental and health concerns. Chemical fate models predict their concentrations using estimates of mass loading, dilution and in-stream attenuation. However, current comprehension of the attenuation rates remains a limiting factor for predictive models. We assessed in-stream attenuation of 75 pharmaceuticals in 4 river segments, aiming to characterize in-stream attenuation variability among different pharmaceutical compounds, as well as among river segments differing in environmental conditions. Our study revealed that in-stream attenuation was highly variable among pharmaceuticals and river segments and that none of the considered pharmaceutical physicochemical and molecular properties proved to be relevant in determining the mean attenuation rates. Instead, the octanol-water partition coefficient (Kow) influenced the variability of rates among river segments, likely due to its effect on sorption to sediments and suspended particles, and therefore influencing the balance between the different attenuation mechanisms (biotransformation, photolysis, sorption, and volatilization). The magnitude of the measured attenuation rates urges scientists to consider them as important as dilution when aiming to predict concentrations in freshwater ecosystems.

Multiresidue trace analysis of sulfonamide antibiotics and their metabolites in soils and sewage sludge by pressurized liquid extraction followed by liquid chromatography-electrospray-quadrupole linear ion trap mass spectrometry.

The present study describes the development, validation and a practical application of a fully automated analytical method based on pressurized liquid extraction (PLE) followed by solid-phase extraction-liquid chromatography-tandem mass spectrometry (SPE-LC-MS/MS) for the simultaneous determination of 22 sulfonamides, including five acetylated metabolites, in sewage sludge and soil samples. Both matrix matched calibration curves and standard calibration curves were built in order to evaluate the potential matrix effects during analysis, and different internal standards were used to compensate these effects during quantification. The recovery efficiencies were found to be 60-130% for the majority of the sulfonamides in both matrices and at two spike levels. The intra-day and inter-day precisions, expressed by the relative standard deviation (RSD), were below 23%. The method limits of detection (MLODs) achieved were in the range 0.03-2.23 ng g(-1) for sewage sludge and 0.01-4.19 ng g(-1) for soil samples. The methodology was applied to evaluate the occurrence of the target sulfonamides in several sewage sludge and soil samples taken in different wastewater treatment plants (WWTPs) and agricultural areas. Results confirmed the wide presence of sulfonamides in both matrices, being sulfathiazole and sulfamethazine the sulfonamides most frequently detected in sewage sludge and soil samples, respectively. Maximum concentrations corresponded to sulfamethazine in both cases (139.2 ng g(-1) and 8.53 ng g(-1) for sewage sludge and soils respectively). Levels were generally lower in soils. Three of the five acetylated metabolites were detected in sewage sludge and two of them in soils, at concentrations not higher than 9.81 ng g(-1).

Ecotoxicity evaluation and removal of sulfonamides and their acetylated metabolites during conventional wastewater treatment.

The present study describes the evaluation of the risk posed by the occurrence of sulfonamides (SAs) in wastewaters. A fully automated analytical method based on on-line solid-phase extraction-liquid chromatography-tandem mass spectrometry (on line SPE-LC-MS/MS) was developed, validated and applied for the analysis of sixteen SAs and, for the first time in wastewaters and sewage sludge, five of their acetylated metabolites. Influent and effluent samples from twenty two different wastewater treatment plants (WWTPs) in Catalonia (Spain) and sewage sludge from fifteen of them were analyzed; removal rates (RE%) and half-lives (t(1/2)) for each SA were calculated. The estimated correlations between RE% and to the hydraulic retention time (HRT) of the different plants indicated no clear influence of HRT on removals. Sulfamethoxazole (SMX), sulfapyridine (SPY) and their corresponding acetylated metabolites were detected with the highest frequencies of detection and at the highest concentrations. The ecotoxicity of both SPY and AcSPY was evaluated for the first time through bioluminescent inhibition assays, resulting in a higher toxicity being attributed to the metabolite. Finally, the potential environmental risk posed by the levels of SAs detected was evaluated calculating the hazard quotients (HQ) to different non-target aquatic organisms in treated wastewaters. SMX was the only SA posing a risk to algae, with an HQ>10.

Removal of sulfonamide antibiotics upon conventional activated sludge and advanced membrane bioreactor treatment.

This work reports the removal efficiencies of nine sulfonamides (SAs) and one of their acetylated metabolites during conventional activated sludge (CAS) and membrane bioreactor (MBR) treatments. Two different types of membranes were studied, hollow-fiber membranes and flat-sheet membranes, in two separate pilot plants operating in parallel to a full-scale CAS treatment. A total of 48 water samples and 16 sewage sludge samples were analyzed by liquid chromatography-tandem mass spectrometry. We obtained 100% elimination in the MBR effluents for three SAs (sulfadiazine, sulfadimethoxine, and sulfamethoxypyridazine) and the metabolite. For the rest of the SAs, the removal efficiencies during CAS and MBR treatments were similar and usually below 55%. Sulfamethizole was the most recalcitrant SA, exhibiting negative removal efficiencies in all the treatments investigated. The concentrations of SAs in the different sewage sludge types were also calculated and ranged from 0.01 to 11 ng g(-1). Furthermore, adsorption and biodegradation of SAs in activated sludge were investigated in two sets of batch reactors, which were spiked at high and low concentration (1,000 and 50 ng mL(-1), respectively). All SAs followed a similar trend and, with the exception of sulfathiazole, were not fully eliminated after 25 days of treatment.

Biodegradation studies of N4-acetylsulfapyridine and N4-acetylsulfamethazine in environmental water by applying mass spectrometry techniques.

This work evaluates the biodegradation of N(4)-acetylsulfapyridine (AcSPY) and N(4)-acetylsulfamethazine (AcSMZ), metabolites of two of the most commonly used sulfonamides (SAs) in human and veterinary medicine, respectively. Aerobic transformation in effluent wastewater was simulated using aerated fixed-bed bioreactors. No visible changes in concentration were observed in the AcSMZ reactor after 90 days, whereas AcSPY was fully degraded after 32 days of experiment. It was also demonstrated that AcSPY transformed back to its parent compound sulfapyridine (SPY). The environmental presence of these two metabolites in wastewater effluent had been previously investigated and confirmed, together with three more SA acetylated metabolites and their corresponding parent compounds, in 18 different wastewater treatment plants in Hesse (Germany). Sulfamethoxazole (SMX) and SPY were the two SAs detected most frequently (90% and 89% of the samples, respectively) and in the highest concentrations (682 ng L(-1) for SMX and 532 ng L(-1) for SPY). To conclude, hazard quotients were calculated whenever toxicity data were available. None of the SAs studied posed an environmental risk.

Kinetic studies and characterization of photolytic products of sulfamethazine, sulfapyridine and their acetylated metabolites in water under simulated solar irradiation.

Sulfapyridine (SPY), sulfonamide (SA) typically used in human therapies, and veterinary SA sulfamethazine (SMZ), are amongst the two SAS most frequently detected in effluent wastewater and surface water respectively. Within this context, this study reports the behaviour of both SAs and their acetylated metabolites, AcSPY and AcSMZ, under artificial irradiance conditions in both high performance liquid chromatography (HPLC) water and in reclaimed wastewater, in order to compare the influence of dissolved organic matter (DOM) and also inorganic matter in the photolysis kinetics. Estimated degradation rate constants (k) ranged from 0.063 h(-1) (SPY) to 2.808 h(-1) (AcSPY), both in HPLC water, with corresponding half-lives (t(1/2)) of 10.93 h and 0.25 h, respectively. A total of 10 different photodegradation products were identified during the photolytic transformation of SPY and 7 for SMZ, through ultra-performance liquid chromatography-quadrupole time of flight mass spectrometry analyses (UPLC-QqTOF-MS), which allowed for exact mass measurements. Regarding the acetylated metabolites, 3 photoproducts were generated for AcSMZ and one for AcSPY. The desulfonated products of each of the four analytes under study were the most relevant photodegradation products identified.