The phototrophic metabolic behaviour of Candidatus accumulibacter.

The phototrophic capability of Candidatus Accumulibacter (Accumulibacter), a common polyphosphate accumulating organism (PAO) in enhanced biological phosphorus removal (EBPR) systems, was investigated in this study. Accumulibacter is phylogenetically related to the purple bacteria Rhodocyclus from the family Rhodocyclaceae, which belongs to the class Betaproteobacteria. Rhodocyclus typically exhibits both chemoheterotrophic and phototrophic growth, however, limited studies have evaluated the phototrophic potential of Accumulibacter. To address this gap, short and extended light cycle tests were conducted using a highly enriched Accumulibacter culture (95%) to evaluate its responses to illumination. Results showed that, after an initial period of adaptation to light conditions (approximately 4-5 h), Accumulibacter exhibited complete phosphorus (P) uptake by utilising polyhydroxyalkanoates (PHA), and additionally by consuming glycogen, which contrasted with its typical aerobic metabolism. Mass, energy, and redox balance analyses demonstrated that Accumulibacter needed to employ phototrophic metabolism to meet its energy requirements. Calculations revealed that the light reactions contributed to the generation of, at least more than 67% of the ATP necessary for P uptake and growth. Extended light tests, spanning 21 days with dark/light cycles, suggested that Accumulibacter generated ATP through light during initial operation, however, it likely reverted to conventional anaerobic/aerobic metabolism under dark/light conditions due to microalgal growth in the mixed culture, contributing to oxygen production. In contrast, extended light tests with an enriched Tetrasphaera culture, lacking phototrophic genes in its genome, clearly demonstrated that phototrophic P uptake did not occur. These findings highlight the adaptive metabolic capabilities of Accumulibacter, enabling it to utilise phototrophic pathways for energy generation during oxygen deprivation, which holds the potential to advance phototrophic-EBPR technology development.

Polyhydroxyalkanoates production in purple phototrophic bacteria ponds: A breakthrough in outdoor pilot-scale operation.

The versatile capacity of purple phototrophic bacteria (PPB) for producing valuable bioproducts has gathered renewed interest in the field of resource recovery and waste valorisation. However, greater knowledge regarding the viability of applying PPB technologies in outdoor, large-scale systems is required. This study assessed, for the first time, the upscaling of the phototrophic polyhydroxyalkanoate (PHA) production technology in a pilot-scale system operated in outdoor conditions. An integrated system composed of two up-flow anaerobic sludge blanket (UASB) reactors (for fermentation of wastewater with molasses), and two high-rate algal ponds retrofitted into PPB ponds, was operated in a wastewater treatment plant under outdoor conditions. UASB's adaptation to the outdoor temperatures involved testing different operational settings, namely hydraulic retention times (HRT) of 48 and 72 h, and molasses fermentation in one or two UASBs. Results have shown that the fermentation of molasses in both UASBs with an increased HRT of 72 h was able to ensure a suitable operation during colder conditions, achieving 3.83 ± 0.63 g CODFermentative Products/L, compared to the 3.73 ± 0.85 g CODFermentative Products/L achieved during warmer conditions (molasses fermentation in one UASB; HRT 48 h). Furthermore, the PPB ponds were operated under a light-feast/dark-aerated-famine strategy and fed with the fermented wastewater and molasses from the two UASBs. The best PHA production was obtained during the summer of 2018 and spring of 2019, attaining 34.7 % gPHA/gVSS with a productivity of 0.11 gPHA L-1 day-1 and 36 % gPHA/gVSS with a productivity of 0.14 gPHA L-1 day-1, respectively. Overall, this study showcases the first translation of phototrophic PHA production technology from an artificially illuminated laboratory scale system into a naturally illuminated, outdoor, pilot-scale system. It also addresses relevant process integration aspects with UASBs for pre-fermenting wastewater with molasses, providing a novel operational strategy to achieve photosynthetic PHA production in outdoor full-scale systems.

Ammonia impact on the selection of a phototrophic - chemotrophic consortium for polyhydroxyalkanoates production under light-feast / dark-aerated-famine conditions.

Phototrophic polyhydroxyalkanoate (PHA) production is an emerging technology for recovering carbon and nutrients from diverse wastewater streams. However, reliable selection methods for the enrichment of PHA accumulating purple phototrophic bacteria (PPB) in phototrophic mixed cultures (PMC) are needed. This research evaluates the impact of ammonia on the selection of a PHA accumulating phototrophic-chemotrophic consortium, towards the enrichment of PHA accumulating PPB. The culture was operated under light-feast/dark-aerated-famine and winter simulated-outdoor conditions (13.2 ± 0.9 °C, transient light, 143.5 W/m2), using real fermented domestic wastewater with molasses as feedstock. Three ammonia supply strategies were assessed: 1) ammonia available only in the light phase, 2) ammonia always present and 3) ammonia available only during the dark-aerated-famine phase. Results showed that the PMC selected under 1) ammonia only in the light and 3) dark-famine ammonia conditions, presented the lowest PHA accumulation capacity during the light period (11.1 % g PHA/g VSS and 10.4 % g PHA/g VSS, respectively). In case 1), the absence of ammonia during the dark-aerated-famine phase did not promote the selection of PHA storing PPB, whereas in case 3) the absence of ammonia during the light period favoured cyanobacteria growth as well as purple sulphur bacteria with increased non-PHA inclusions, resulting in an overall decrease of phototrophic PHA accumulation capacity. The best PHA accumulation performance was obtained with selection under permanent presence of ammonia (case 2), which attained a PHA content of 21.6 % g PHA/g VSS (10.2 Cmmol PHA/L), at a production rate of 0.57 g PHA/L·day, during the light period in the selection reactor. Results in case 2 also showed that feedstock composition impacts the PMC performance, with feedstocks richer in more reduced volatile fatty acids (butyric and valeric acids) decreasing phototrophic performance and leading to acids entering the dark-aerated phase. Nevertheless, the presence of organic carbon in the aerated phase was not detrimental to the system. In fact, it led to the establishment of a phototrophic-chemotrophic consortium that could photosynthetically accumulate a PHA content of 13.2 % g PHA/g VSS (6.7 Cmmol PHA/L) at a production rate of 0.20 g PHA/L·day in the light phase, and was able to further increase that storage up to 18.5 % g PHA/g VSS (11.0 Cmmol PHA/L) at a production rate of 1.35 g PHA/L·day in the dark-aerated period. Furthermore, the light-feast/dark-aerated-famine operation was able to maintain the performance of the selection reactor under winter conditions, unlike non-aerated PMC systems operated under summer conditions, suggesting that night-time aeration coupled with the constant presence of ammonia can contribute to overcoming the seasonal constraints of outdoor operation of PMCs for PHA production.

Long term operation of a phototrophic biological nutrient removal system: Impact of CO2 concentration and light exposure on process performance.

The utilization of non-aerated microalgae-bacterial consortia for phototrophic biological nutrient removal (photo-BNR) has emerged as an alternative to conventional wastewater treatment. Photo-BNR systems are operated under transient illumination, with alternating dark-anaerobic, light-aerobic and dark-anoxic conditions. A deep understanding of the impact of operational parameters on the microbial consortium and respective nutrient removal efficiency in photo-BNR systems is required. The present study evaluates, for the first time, the long-term operation (260 days) of a photo-BNR system, fed with a COD:N:P mass ratio of 7.5:1:1, to understand its operational limitations. In particular, different CO2 concentrations in the feed (between 22 and 60 mg C/L of Na2CO3) and variations of light exposure (from 2.75 h to 5.25 h per 8 h cycle) were studied to determine their impact on key parameters, like oxygen production and availability of polyhydroxyalkanoates (PHA), on the performance of anoxic denitrification by polyphosphate accumulating organisms. Results indicate that oxygen production was more dependent on the light availability than on the CO2 concentration. Also, under operational conditions with a COD:Na2CO3 ratio of 8.3 mg COD/mg C and an average light availability of 5.4 ± 1.3 W h/g TSS, no internal PHA limitation was observed, and 95 ± 7%, 92 ± 5% and 86 ± 5% of removal efficiency could be achieved for phosphorus, ammonia and total nitrogen, respectively. 81 ± 1.7% of the ammonia was assimilated into the microbial biomass and 19 ± 1.7% was nitrified, showing that biomass assimilation was the main N removal mechanism taking place in the bioreactor. Overall, the photo-BNR system presented a good settling capacity (SVI ∼60 mL/g TSS) and was able to remove 38 ± 3.3 mg P/L and 33 ± 1.7 mg N/L, highlighting its potential for achieving wastewater treatment without the need of aeration.

Achieving nitrogen and phosphorus removal at low C/N ratios without aeration through a novel phototrophic process.

Conventional wastewater treatment technologies for biological nutrient removal (BNR) are highly dependent on aeration for oxygen supply, which represents a major operational cost of the process. Recently, phototrophic enhanced biological phosphorus removal (photo-EBPR) has been suggested as an alternative system for phosphorus removal, based on a consortium of photosynthetic microorganisms and chemotrophic bacteria, eliminating the need for costly aeration. However, wastewater treatment plants must couple nitrogen and phosphorus removal to achieve discharge limits. For this reason, a new microalgae-bacterial based system for phosphorus and nitrogen removal is proposed in this work. The photo-BNR system studied here consists of a sequencing batch reactor operated with dark anaerobic, light aerobic, dark anoxic and idle periods, to allow both N and P removal. Results of the study show that the photo-BNR system was able to remove 100% of the 40 mg N/L of ammonia fed to the reactor and 94 ± 3% of the total nitrogen (Influent COD:N ratio of 300:40, similar to domestic wastewater). Moreover, an average of 25 ± 9.2 mg P/L was simultaneously removed in the photo-BNR tests, representing the P removal capacity of this system, which exceeds the level of P removal required from typical domestic wastewater. Full ammonia removal was achieved during the light phase, with 67 ± 5% of this ammonia being assimilated by the microbial culture and the remaining 33 ± 5% being converted into nitrate. The assimilated P corresponded to 2.8 ± 0.23 mg P/L, which only represented, approximately, 1/9 of the P removal capacity of the system. Half of the nitrified ammonia was subsequently denitrified during the dark anoxic phase (50 ± 24%). Overall, the photo-BNR system represents the first treatment alternative for N and P from domestic wastewater with no need of mechanical aeration or supplemental carbon addition, representing an alternative low-energy technology of interest.

Polyhydroxyalkanoates production from fermented domestic wastewater using phototrophic mixed cultures.

Phototrophic mixed cultures (PMC) have been found to be a promising technology to produce polyhydroxyalkanoates (PHA), however, work performed thus far has focussed mainly on the use of synthetic feedstocks and operational conditions that differ from those expectable in full-scale processes. The goals of this work were to study, for the first time, the capability of PMCs to produce PHA using real fermented domestic wastewater as feedstock under mixing/light/temperature conditions that are naturally found in outdoor open systems. Various operational strategies were evaluated in this study to increase PHA productivity, namely the poly(3-hydroxybutyric-co-3-hydroxyvaleric) copolymer (PHBV) by PMC systems. Two lab-scale photobioreactors were operated in parallel, with transient illumination (12 h light/12 h dark) and subjected to feedstock fluctuations under two culture selection strategies that best suit the oxidative conditions of high rate algal ponds (HRAPs) which are commonly applied in wastewater treatment plants (WWTP). Under a permanent carbon feast regime (selection strategy 1), the PMC became highly enriched in phototrophic purple bacteria (PPB), and two complementary conditions that can improve the selection of PHA accumulating bacteria were discovered: phosphate cycling, where 20% PHA/VSS (86HB:14HV in a C-mol basis) with a light phase productivity of 0.23 g PHA/L•d_light phase was attained; and transitioning from selection under low organic loading rate (OLR) to high OLR where 17.6% PHA/VSS (60HB:40HV in C base) with a light phase productivity of 0.18 g PHA/L•d_light phase was achieved. Under a feast and famine regime (selection strategy 2), a PMC consortium of microalgae and PPB was obtained, and a multiple pulse feeding strategy during the first hours of the light phase in the selector reactor led to a 26.1% PHA/VSS (36HB:64HV in C base) content, with a productivity of 0.26 g PHA/L•d_light phase and 0.52 g PHA/L•d_feast phase. An accumulation test under higher light intensity led to 30.8% PHA/VSS (85HB:15HV on a C-mol basis) with a productivity of 2.67 g PHA/L•d, along the 8 h of accumulation.

Metabolic modelling of mixed culture anaerobic microbial processes.

Mixed culture anaerobic processes are important to environmental systems, including the global carbon cycle, and industrial and environmental biotechnology. Mixed culture metabolic modelling (MM) is an essential tool to analyse these systems. MM predicts microbial function based on knowledge or assumption of cellular metabolism. It may be developed based on observations at the process level - biochemical process modelling (BPM) or fundamental knowledge of the cell being modelled - cellular level modelling (CLM). There is a substantial gap between these two fields, with BPM not considering genetic constraints, particularly where this may be important to interspecies interactions (e.g. amino acid transfer), and CLM commonly not considering mass transfer principles, such as advection/diffusion/migration. No unified approach is useful for all applications, but there is an increasing need to consider genetic information and constraints in developing BPM, and translate BPM principles (including mass-transfer and inorganic chemistry) for application to CLM.

Improving polyhydroxyalkanoates production in phototrophic mixed cultures by optimizing accumulator reactor operating conditions.

Polyhydroxyalkanoates (PHAs) production with phototrophic mixed cultures (PMCs) has been recently proposed. These cultures can be selected under the permanent presence of carbon and the PHA production can be enhanced in subsequent accumulation steps. To optimize the PHA production in accumulator reactors, this work evaluated the impact of 1) initial acetate concentration, 2) light intensity, 3) removal of residual nitrogen on the culture performance. Results indicate that low acetate concentration (<30 CmM) and specific light intensities around 20 W/gX are optimal operating conditions that lead to high polyhydroxybutyrate (PHB) storage yields (0.83 ±â€¯0.07 Cmol-PHB/Cmol-Acet) and specific PHB production rates of 2.21 ±â€¯0.07 Cmol-PHB/Cmol X d. This rate is three times higher than previously registered in non-optimized accumulation tests and enabled a PHA content increase from 15 to 30% in <4 h. Also, it was shown for the first time, the capability of a PMC to use a real waste, fermented cheese whey, to produce PHA with a hydroxyvalerate (HV) content of 12%. These results confirm that fermented wastes can be used as substrates for PHA production with PMCs and that the energy levels in sunlight that lead to specific light intensities from 10 to 20 W/gX are sufficient to drive phototrophic PHA production processes.

The link between nitrous oxide emissions, microbial community profile and function from three full-scale WWTPs.

Few attempts have been made in previous studies to link the microbial community structure and function with nitrous oxide (N2O) emissions at full-scale wastewater treatment plants (WWTPs). In this work, high-throughput sequencing and reverse transcriptase-qPCR (RT-qPCR) was applied to activated sludge samples from three WWTPs for two seasonal periods (winter and summer) and linked with the N2O emissions and wastewater characteristics. The total N2O emissions ranged from 7.2 to 937.0 g N-N2O/day, which corresponds to an emission factor of 0.001 to 0.280% of the influent NH4-N being emitted as N2O. Those emissions were related to the abundance of Nitrotoga, Candidatus Microthrix and Rhodobacter genera, which were favored by higher dissolved oxygen (DO) and nitrate (NO3-) concentrations in the activated sludge tanks. Furthermore, a relationship between the nirK gene expression and N2O emissions was verified. Detected N2O emission peaks were associated with different process events, related to aeration transition periods, that occurred during the regular operation of the plants, which could be potentially associated to increased emissions of the WWTP. The design of mitigation strategies, such as optimizing the aeration regime, is therefore important to avoid process events that lead to those N2O emissions peaks. Furthermore, this study also demonstrates the importance of assessing the gene expression of nosZ clade II, since its high abundance in WWTPs could be an important key to reduce the N2O emissions.

The effect of seed sludge on the selection of a photo-EBPR system.

The phototrophic-enhanced biological phosphorus removal system (photo-EBPR) was recently proposed as an alternative photosynthetic process to conventional phosphorus removal. Previous work showed the possibility of obtaining a photo-EBPR system starting from a culture already enriched in polyphosphate accumulating organisms (PAOs). The present work evaluated whether the same could be achieved starting from conventional activated sludge. A sequencing batch reactor inoculated with sludge from a wastewater treatment plant (WWTP) was fed with a mixture of acetate and propionate (75%:25%) and subjected to dark/light cycles to select a photo-EBPR system containing PAOs and photosynthetic organisms, the oxygen providers for the system. The results showed that it is possible to obtain a photo-EBPR system starting from a WWTP sludge, although the process is slower than when started with a sludge already enriched in PAOs. After 15 days of operation, the system could remove 60 ± 2 mg-P/L of phosphorus (approximately 67% of the concentration at the end of dark period) in the light period, from which 13 ± 1 mg-P/L was removed during the phase without external air supply. These results indicate that a photo-EBPR system can be obtained independently of the seed sludge initially used, provided that a suitable operating strategy is implemented, i.e. by imposing conditions that favour the growth and coexistence of PAOs and photosynthetic microorganisms.

The link between the microbial ecology, gene expression, and biokinetics of denitrifying polyphosphate-accumulating systems under different electron acceptor combinations.

The emission of the greenhouse gas nitrous oxide (N2O) can occur during biological nutrient removal. Denitrifying enhanced biological phosphorus removal (d-EBPR) systems are an efficient means of removing phosphate and nitrogen, performed by denitrifying polyphosphate-accumulating organisms (d-PAOs). The aim of this work was to study the effect of various combinations of electron acceptors, nitrate (NO3-), nitrite (NO2-), and N2O, on the denitrification pathway of a d-EBPR system. Batch tests were performed with different electron acceptor combinations, to explore the denitrification pathway. Reverse transcriptase-qPCR (RT-qPCR) and high-throughput sequencing, combined with chemical analysis, were used to study gene expression, microbial diversity, and denitrification kinetics. The potential for N2O production was greater than the potential for its reduction in most tests. A strong correlation was observed between the N2O reduction rate and the relative gene expression of nitrous oxide reductase per nitrite reductase (nosZ/(nirS + nirK)), suggesting that the expression of denitrifying marker genes is a strong predictor of the N2O reduction rate. The d-EBPR community maintained a core population with low variations throughout the study. Furthermore, phylogenetic analyses of the studied marker genes revealed that the organisms actively involved in denitrification were closely related to Thauera sp., Candidatus Accumulibacter phosphatis, and Candidatus Competibacter denitrificans. Moreover, Competibacter-related OTUs seem to be important contributors to the N2O reduction capacity of the system, likely scavenging the N2O produced by other organisms. Overall, this study contributes to a better understanding of the microbial biochemistry and the genetics involving biological denitrification removal, important to minimize N2O emissions in wastewater treatment plants.

The impact of operational strategies on the performance of a photo-EBPR system.

A novel Phototrophic - Enhanced Biological Phosphorus Removal (Photo-EBPR) system, consisting of a consortium of photosynthetic organisms and polyphosphate accumulating organisms (PAOs), was studied in this work. A sequencing batch reactor was fed with a mixture of acetate and propionate (75%-25%) and subjected to dark/light cycles in order to select a photo-EBPR system containing PAOs and photosynthetic organisms, the latter likely providers of oxygen to the system. The results from the selection period (stage 1) showed that the photo-EBPR culture was capable of performing P release in the dark and P uptake in the presence of light, under limited oxygen concentrations. During the optimization period, the aeration period, which was initially provided at the end of the light phase, was gradually reduced until a non-aerated system was achieved, while the light intensity was increased. After optimization of the operational conditions, the selected consortium of photosynthetic organisms/PAOs showed high capacity of P removal in the light phase in the absence of air or other electron acceptor. A net P removal of 34 ± 3 mg-P/L was achieved, with a volumetric P removal rate of 15 ± 2 mg-P/L.h, and 79 ± 8% of P removal from the system. Also, in limiting oxygen conditions, the P uptake rate was independent of the PHA consumption, which demonstrates that the organisms obtained energy for P removal from light. These results indicated that a photo-EBPR system can be a potential solution for P removal with low COD/P ratios and in the absence of air, prospecting the use of natural sunlight as illumination, which would reduce the costs of EBPR operation regarding aeration.

Modelling energy costs for different operational strategies of a large water resource recovery facility.

The main objective of this paper is to demonstrate the importance of applying dynamic modelling and real energy prices on a full scale water resource recovery facility (WRRF) for the evaluation of control strategies in terms of energy costs with aeration. The Activated Sludge Model No. 1 (ASM1) was coupled with real energy pricing and a power consumption model and applied as a dynamic simulation case study. The model calibration is based on the STOWA protocol. The case study investigates the importance of providing real energy pricing comparing (i) real energy pricing, (ii) weighted arithmetic mean energy pricing and (iii) arithmetic mean energy pricing. The operational strategies evaluated were (i) old versus new air diffusers, (ii) different DO set-points and (iii) implementation of a carbon removal controller based on nitrate sensor readings. The application in a full scale WRRF of the ASM1 model coupled with real energy costs was successful. Dynamic modelling with real energy pricing instead of constant energy pricing enables the wastewater utility to optimize energy consumption according to the real energy price structure. Specific energy cost allows the identification of time periods with potential for linking WRRF with the electric grid to optimize the treatment costs, satisfying operational goals.

Impact of biogenic substrates on sulfamethoxazole biodegradation kinetics by Achromobacter denitrificans strain PR1.

Pure cultures have been found to degrade pharmaceutical compounds. However, these cultures are rarely characterized kinetically at environmentally relevant concentrations. This study investigated the kinetics of sulfamethoxazole (SMX) degradation by Achromobacter denitrificans strain PR1 at a wide range of concentrations, from ng/L to mg/L, to assess the feasibility of using it for bioaugmentation purposes. Complete removal of SMX occurred for all concentrations tested, i.e., 150 mg/L, 500 µg/L, 20 µg/L, and 600 ng/L. The reaction rate coefficients (kbio) for the strain at the ng/L SMX range were: 63.4 ± 8.6, 570.1 ± 15.1 and 414.9 ± 124.2 L/g[Formula: see text]·day), for tests fed without a supplemental carbon source, with acetate, and with succinate, respectively. These results were significantly higher than the value reported for non-augmented activated sludge (0.41 L/(g [Formula: see text]·day) with hundreds of ng/L of SMX. The simultaneous consumption of an additional carbon source and SMX suggested that the energetic efficiency of the cells, boosted by the presence of biogenic substrates, was important in increasing the SMX degradation rate. The accumulation of 3-amino-5-methylisoxazole was observed as the only metabolite, which was found to be non-toxic. SMX inhibited the Vibrio fischeri luminescence after 5 min of contact, with EC50 values of about 53 mg/L. However, this study suggested that the strain PR1 still can degrade SMX up to 150 mg/L. The results of this work demonstrated that SMX degradation kinetics by A. denitrificans PR1 compares favorably with activated sludge and the strain is a potentially interesting organism for bioaugmentation for SMX removal from polluted waters.

Beyond feast and famine: Selecting a PHA accumulating photosynthetic mixed culture in a permanent feast regime.

Currently, the feast and famine (FF) regime is the most widely applied strategy to select for polyhydroxyalkanoate (PHA) accumulating organisms in PHA production systems with mixed microbial cultures. As an alternative to the FF regime, this work studied the possibility of utilizing a permanent feast regime as a new operational strategy to select for PHA accumulating photosynthetic mixed cultures (PMCs). The PMC was selected in an illuminated environment and acetate was constantly present in the mixed liquor to guarantee a feast regime. During steady-state operation, the culture presented low PHA accumulation levels, likely due to low light availability, which resulted in most of the acetate being used for biomass growth (Yx/s of 0.64 ± 0.18 Cmol X/Cmol Acet). To confirm the light limitation on the PMC, SBR tests were conducted with higher light availability, at similar levels as would be expectable from natural sunlight. In this case, the Yx/s reduced to 0.11 ± 0.01 Cmol X/Cmol Acet and the culture presented a PHB production yield on acetate of 0.67 ± 0.01 Cmol PHB/Cmol Acet, leading to a maximum PHB content of 60%. Unlike other studied PMCs, the PMC was capable of simultaneous growth and PHB accumulation continuously throughout the cycle. Thus far, 60% PHA content is the maximum value ever reported for a PMC, a result that prospects the utilization of feast regimes as an alternative strategy for the selection of PHA accumulating PMCs. Furthermore, the PMC also presented high phosphate removal rates, delivering an effluent that complies with phosphate discharge limits. The advantages of selecting PMCs under a permanent feast regime are that no aeration inputs are required; it allows higher PHA contents and phosphate removal rates in comparison to FF-operated PMC systems; and it represents a novel means of integrating wastewater treatment with resource recovery in the form of PHA.

Status of hormones and painkillers in wastewater effluents across several European states-considerations for the EU watch list concerning estradiols and diclofenac.

Present technologies for wastewater treatment do not sufficiently address the increasing pollution situation of receiving water bodies, especially with the growing use of personal care products and pharmaceuticals (PPCP) in the private household and health sector. The relevance of addressing this problem of organic pollutants was taken into account by the Directive 2013/39/EU that introduced (i) the quality evaluation of aquatic compartments, (ii) the polluter pays principle, (iii) the need for innovative and affordable wastewater treatment technologies, and (iv) the identification of pollution causes including a list of principal compounds to be monitored. In addition, a watch list of 10 other substances was recently defined by Decision 2015/495 on March 20, 2015. This list contains, among several recalcitrant chemicals, the painkiller diclofenac and the hormones 17ß-estradiol and 17α-ethinylestradiol. Although some modern approaches for their removal exist, such as advanced oxidation processes (AOPs), retrofitting most wastewater treatment plants with AOPs will not be acceptable as consistent investment at reasonable operational cost. Additionally, by-product and transformation product formation has to be considered. The same is true for membrane-based technologies (nanofiltration, reversed osmosis) despite of the incredible progress that has been made during recent years, because these systems lead to higher operation costs (mainly due to higher energy consumption) so that the majority of communities will not easily accept them. Advanced technologies in wastewater treatment like membrane bioreactors (MBR) that integrate biological degradation of organic matter with membrane filtration have proven a more complete elimination of emerging pollutants in a rather cost- and labor-intensive technology. Still, most of the presently applied methods are incapable of removing critical compounds completely. In this opinion paper, the state of the art of European WWTPs is reflected, and capacities of single methods are described. Furthermore, the need for analytical standards, risk assessment, and economic planning is stressed. The survey results in the conclusion that combinations of different conventional and advanced technologies including biological and plant-based strategies seem to be most promising to solve the burning problem of polluting our environment with hazardous emerging xenobiotics.

Ecotoxicity of ketoprofen, diclofenac, atenolol and their photolysis byproducts in zebrafish (Danio rerio).

The occurrence of pharmaceutical compounds in wastewater treatment plants and surface waters has been detected worldwide, constituting a potential risk for aquatic ecosystems. Adult zebrafish, of both sexes, were exposed to three common pharmaceutical compounds (atenolol, ketoprofen and diclofenac) and their UV photolysis by-products over seven days. The results show that diclofenac was removed to concentrations

Modelling the metabolic shift of polyphosphate-accumulating organisms.

Enhanced biological phosphorus removal (EBPR) is one of the most important methods of phosphorus removal in municipal wastewater treatment plants, having been described by different modelling approaches. In this process, the PAOs (polyphosphate accumulating organisms) and GAOs (glycogen accumulating organisms) compete for volatile fatty acids uptake under anaerobic conditions. Recent studies have revealed that the metabolic pathways used by PAOs in order to obtain the energy and the reducing power needed for polyhydroxyalkanoates synthesis could change depending on the amount of polyphosphate stored in the cells. The model presented in this paper extends beyond previously developed metabolic models by including the ability of PAO to change their metabolic pathways according to the content of poly-P available. The processes of the PAO metabolic model were adapted to new formulations enabling the change from P-driven VFA uptake to glycogen-driven VFA uptake using the same process equations. The stoichiometric parameters were changed from a typical PAO coefficient to a typical GAO coefficient depending on the internal poly-P with Monod-type expressions. The model was calibrated and validated with seven experiments under different internal poly-P concentrations, showing the ability to correctly represent the PAO metabolic shift at low poly-P concentrations. The sensitivity and error analysis showed that the model is robust and has the ability to describe satisfactorily the change from one metabolic pathway to the other one, thereby encompassing a wider range of process conditions found in EBPR plants.

Photosynthetic mixed culture polyhydroxyalkanoate (PHA) production from individual and mixed volatile fatty acids (VFAs): substrate preferences and co-substrate uptake.

This work studied the effect of the substrate feeding composition on the polyhydroxyalkanoate (PHA) accumulation capacity of an acetate enriched photosynthetic mixed culture (PMC). From the six tested organic acids - malate, citrate, lactate, acetate, propionate and butyrate - only the three volatile fatty acids (VFAs) enabled PHA production, with acetate and butyrate leading to polyhydroxybutyrate (PHB) formation and propionate leading to a HB:HV copolymer with a 51% fraction of hydroxyvalerate (HV). Also, results showed an acceleration of butyrate and propionate consumption when fed in the presence of acetate, suggesting that the latter can act as a co-substrate for butyrate and propionate uptake. Furthermore, results suggest that some PMC bacterial groups present a substrate preference for butyrate in relation to acetate and propionate. These findings indicate the possibility of feeding the PMC with cheap VFA rich fermented wastes, leading to a more cost-effective and environmentally sustainable PHA production system.

Effect of dark/light periods on the polyhydroxyalkanoate production of a photosynthetic mixed culture.

This work studied the possibility of operating a viable polyhydroxyalkanoate (PHA) producing photosynthetic mixed culture (PMC) under dark/light periods without aeration. The culture was subjected to a feast and famine regime, being fed in the dark phase and entering into famine during the light phase. Throughout consecutive feast and famine dark/light periods, the PMC became enriched in PHA accumulating organisms, where non-PHA producing algae that can grow under continuous illumination were out-competed. The very low algae levels enabled greater light and carbon source availability for PHA accumulating bacteria, leading to higher metabolic rates and PHA levels. The PMC reached a PHA content of 30% PHA/VSS, and doubled its specific PHA production rate in relation to PMCs operated previously under continuous illumination. This new process takes a further step towards operating a more cost effective PMC system for PHA production, opening up the possibility for direct sunlight utilization in the future.