Modified Poly(acrylic acid)-Based Hydrogels for Enhanced Mainstream Removal of Ammonium from Domestic Wastewater.

Rapid and continuous ammonium adsorption from mainstream coupled with side-stream ammonium recovery and adsorbent regeneration could enable ammonium recovery from domestic wastewater. This study describes the use of tailored poly(acrylic acid)-based (NaPAA) hydrogels as effective sorbents for ammonium removal from domestic wastewater. Modified NaPAA hydrogels having 60% ionization and 4.8 mol % N',N'-methylenebisacrylamide as the cross-linker reduced the overall swelling by 92% from 407 to 31 g/g because of higher cross-linking density. At hydrogel loadings of 2.5-7.5 g/L, the NaPAA hydrogels achieved ammonium concentrations of 8.3 ± 0.6 to 10.1 ± 0.1 mg/L NH4-N, which corresponds to removal efficiencies of 53-77% after 10 min of contact time in real domestic wastewater. At the same hydrogel loadings, the ammonium removal efficiency of NaPAA hydrogels in synthetic wastewater was found to be comparable to that in real sewage (71% vs 69%, respectively), suggesting that the sorption performance is only marginally affected by organic constituents found in domestic wastewater. In addition, the NaPAA hydrogels removed 25-51% ammonium in 10 min from synthetic streams having 200-400% higher ionic strengths than those commonly observed in sewage. Furthermore, simulation studies showed that a discharge concentration of ∼1.9 mg/L NH4-N, well below the commonly applied discharge limits in most regions, can be achieved using mainstream ammonium removal by NaPAA hydrogels followed by biological assimilation from the growth of ordinary heterotrophic organisms.

The impact of temperature on the metabolism of volatile fatty acids by polyphosphate accumulating organisms (PAOs).

This study investigated the effects of different carbon sources on enriched Accumulibacter PAO cultures at high temperature (30 °C) and compared the carbon transformation with low temperature (20 °C) cases reported in literature, revealing several key metabolic differences. While PAOs seemed to prefer propionate anaerobically as compared to other VFAs at high temperature, high aerobic glycogen replenishment was realized with propionate as the anaerobic carbon source, a trait not previously observed at low temperatures. Therefore, it was found that propionate is not correlated with high P removal by Accumulibacter PAO at high temperatures. A combined substrate of acetate, propionate and perhaps butyrate seemed to be a better carbon source combination, since the total VFA uptake rate increased by up to 46%, and this increased the aerobic P-removal efficiency by up to 38.4% and reduced the glycogen recovery by more than 63% compared to the use of only propionate as substrate. This study improves our understanding of how to stimulate successful EBPR operation in warm climates by augmenting the P removal performance of PAOs.

Oerskovia paurometabola can efficiently decolorize azo dye Acid Red 14 and remove its recalcitrant metabolite.

The biodegradation of dyes remains one of the biggest challenges of textile wastewater. Azo dyes are one of the most commonly employed dye classes, and biological treatment processes tend to generate recalcitrant aromatic amines, which are more toxic than the parent dye molecule. This study aimed to isolate bacterial strains with the capacity to degrade both the azo dye and the resulting aromatic amines towards the development of a simple and reliable treatment approach for dye-laden wastewaters. A mixed bacterial enrichment was first developed in an anaerobic-aerobic lab-scale sequencing batch reactor (SBR) fed with a synthetic textile wastewater containing the model textile azo dye Acid Red 14 (AR14). Eighteen bacterial strains were isolated from the SBR, including members of the Acinetobacter, Pseudomonas and Oerskovia genera, Oerskovia paurometabola presenting the highest decolorization capacity (91% after 24 h in static anaerobic culture). Growth assays supported that this is a facultative bacterium, and decolorization batch tests with 20-100 mg AR14 L-1 in a synthetic textile wastewater supplemented with yeast extract indicated that O. paurometabola has a high color removal capacity for a significant range of AR14 concentrations. In addition, a model typically used to describe biodegradation of xenobiotic compounds was adjusted to the results, to predict AR14 biodegradation time profiles at different initial concentrations. HPLC analysis confirmed that decolorization occurred through azo bond reduction under anaerobic conditions, the azo dye being completely reduced after 24 h of anaerobic incubation for the range of concentrations tested. Interestingly, partial (up to 63%) removal of one of the resulting aromatic amines (4-amino-naphthalene-1-sulfonic acid) was observed when subsequently subjected to aerobic conditions. Overall, this work showed the azo dye biodegradation potential of specific bacterial strains isolated from mixed culture bioreactors, reporting for the first time the decolorization capacity of an Oerskovia sp. with further biodegradation of a recalcitrant sulfonated aromatic amine metabolite.

Denitrification activity of polyphosphate accumulating organisms (PAOs) in full-scale wastewater treatment plants.

A comprehensive assessment of full-scale enhanced biological phosphorus removal (EBPR) plants (five plants, 19 independent tests) was undertaken to determine their effectiveness in terms of aerobic and anoxic P removal. By comparing parallel P uptake tests under only aerobic or under anoxic-aerobic conditions, results revealed that introducing an anoxic stage led to an overall P removal of on average 90% of the P removed under only aerobic conditions. This was achieved with negligible higher PHA and glycogen requirements, 30% lower overall oxygen consumption and with the simultaneous removal of nitrate, reducing up to an estimate of 70% of carbon requirements for simultaneous N and P removal. Varying fractions of denitrifying polyphosphate accumulating organisms (DPAOs), from an average of 25% to 84%, were found in different plants. No correlation was found between the DPAO fractions and EBPR configuration, season, or the concentration of any of the microbial groups measured via quantitative fluorescence in situ hybridisation. These included Type I and Type II Ca. Accumulibacter and glycogen accumulating organisms, suggesting that chemical batch tests are the best methodology for quantifying the potential of anoxic P removal in full-scale wastewater treatment plants.

Development of a Novel Process Integrating the Treatment of Sludge Reject Water and the Production of Polyhydroxyalkanoates (PHAs).

Polyhydroxyalkanoates (PHAs) from activated sludge and renewable organic material can become an alternative product to traditional plastics since they are biodegradable and are produced from renewable sources. In this work, the selection of PHA storing bacteria was integrated with the side stream treatment of nitrogen removal via nitrite from sewage sludge reject water. A novel process was developed and applied where the alternation of aerobic-feast and anoxic-famine conditions accomplished the selection of PHA storing biomass and nitrogen removal via nitrite. Two configurations were examined: in configuration 1 the ammonium conversion to nitrite occurred in the same reactor in which the PHA selection process occurred, while in configuration 2 two separate reactors were used. The results showed that the selection of PHA storing biomass was successful in both configurations, while the nitrogen removal efficiency was much higher (almost 90%) in configuration 2. The PHA selection degree was evaluated by the volatile fatty acid (VFA) uptake rate (-qVFAs) and the PHA production rate (qPHA), which were 239 ± 74 and 89 ± 7 mg of COD per gram of active biomass (Xa) per hour, respectively. The characterization of the biopolymer recovered after the accumulation step, showed that it was composed of 3-hydroxybutyrate (3HB) (60%) and 3-hydroxyvalerate (3HV) (40%). The properties associated with the produced PHA suggest that they are suitable for thermoplastic processing.

Novel microelectrode-based online system for monitoring N2O gas emissions during wastewater treatment.

Clark-type nitrous oxide (N2O) microelectrodes are commonly used for measuring dissolved N2O levels, but have not previously been tested for gas-phase applications, where the N2O emitted from wastewater systems can be directly quantified. In this study, N2O microelectrodes were tested and validated for online gas measurements, and assessed with respect to their temperature, gas flow, composition dependence, gas pressure, and humidity. An exponential correlation between temperature and sensor signal was found, whereas gas flow, composition, pressure, and humidity did not have any influence on the signal. Two of the sensors were tested at different N2O concentration ranges (0-422.3, 0-50, 0-10, and 0-2 ppmv N2O) and exhibited a linear response over each range. The N2O emission dynamics from two laboratory scale sequencing batch reactors performing ammonia or nitrite oxidation were also monitored using one of the microsensors and results were compared with two other analytical methods. Results show that N2O emissions were accurately described with these microelectrodes and support their application for assessing gaseous N2O emissions from wastewater treatment systems. Advantages of the sensors as compared to conventional measurement techniques include a wider quantification range of N2O fluxes, and a single measurement system that can assess both liquid and gas-phase N2O dynamics.

Monitoring intracellular polyphosphate accumulation in enhanced biological phosphorus removal systems by quantitative image analysis.

A rapid methodology for intracellular storage polyphosphate (poly-P) identification and monitoring in enhanced biological phosphorus removal (EBPR) systems is proposed based on quantitative image analysis (QIA). In EBPR systems, 4',6-diamidino-2-phenylindole (DAPI) is usually combined with fluorescence in situ hybridization to evaluate the microbial community. The proposed monitoring technique is based on a QIA procedure specifically developed for determining poly-P inclusions within a biomass suspension using solely DAPI by epifluorescence microscopy. Due to contradictory literature regarding DAPI concentrations used for poly-P detection, the present work assessed the optimal DAPI concentration for samples acquired at the end of the EBPR aerobic stage when the accumulation occurred. Digital images were then acquired and processed by means of image processing and analysis. A correlation was found between average poly-P intensity values and the analytical determination. The proposed methodology can be seen as a promising alternative procedure for quantifying intracellular poly-P accumulation in a faster and less labour-intensive way.

Assessment of three different approaches for integrating phosphorus recovery from sewage sludge and derived products in existing wastewater treatment plants.

Three different technological solutions, namely acidogenic fermentation and chemical extraction (alkaline or acidic), followed by precipitation with 1% Ca(OH)2, were investigated in the view of integrating phosphorus recovery into existing wastewater treatment plants. Experiments were conducted at the lab-scale using (i) sludge taken from biologically and chemically promoted phosphorus removal activated sludge processes and (ii) ashes obtained from sludge muffle incineration. Results highlighted the benefits of enhanced biological phosphorus removal (EBPR) systems rather than chemically promoted phosphorus removal in not only phosphorus extraction (up to 40% with EBPR) and recovery directly from secondary sludge (P precipitation between 66 and 92%), but after sludge incineration as well (P extraction up to 96% and precipitation above 96%). Acidogenic fermentation ensured the highest phosphorus release from EBPR sludge (equal to a concentration in solution of 122 mg/L P-PO43-), while the derived ashes had a lower level of metal contamination (particularly Fe and Al content < 2%). The phosphorus-rich product obtained by means of the recovery process showed relevant metal contamination (Cu, Zn, and Ni) under some operating conditions, suggesting the need for further treatments.

Insights on biological phosphorus removal with partial nitrification in single sludge system via sidestream free ammonia and free nitrous acid dosing.

The sidestream sludge treatment by free ammonium (FA)/free nitrous acid (FNA) dosing was frequently demonstrated to maintain the nitrite pathway for the partial nitrification (PN) process. Nevertheless, the inhibitory effect of FA and FNA would severely influence polyphosphate accumulating organisms (PAOs), destroying the microbe-based phosphorus (P) removal. Therefore, a strategic evaluation was proposed to successfully achieve biological P removal with a partial nitrification process in a single sludge system by sidestream FA and FNA dosing. Through the long-term operation of 500 days, excellent phosphorus, ammonium and total nitrogen removal performance were achieved at 97.5 ± 2.6 %, 99.1 ± 1.0 % and 75.5 ± 0.4 %, respectively. Stable partial nitrification with a nitrite accumulation ratio (NAR) of 94.1 ± 3.4 was attained. The batch tests also reported the robust aerobic phosphorus uptake based on FA and FNA adapted sludge after exposure of FA and FNA, respectively, suggesting the FA and FNA treatment strategy could potentially offer the opportunity for the selection of PAOs, which synchronously have the tolerance to FA and FNA. Microbial community analysis suggested that Accumulibacter, Tetrasphaera, and Comamonadaceae collectively contributed to the phosphorus removal in this system. Summarily, the proposed work presents a novel and feasible strategy to integrate enhanced biological phosphorus removal (EBPR) and short-cut nitrogen cycling and bring the combined mainstream phosphorus removal and partial nitrification process closer to practical application.

The impact of pH on the anaerobic and aerobic metabolism of Tetrasphaera-enriched polyphosphate accumulating organisms.

Members of the genus Tetrasphaera are putative polyphosphate accumulating organisms (PAOs) that have been found in greater abundance than Accumulibacter in many full-scale enhanced biological phosphorus removal (EBPR) wastewater treatment plants worldwide. Nevertheless, previous studies on the effect of environmental conditions, such as pH, on the performance of EBPR have focused mainly on the response of Accumulibacter to pH changes. This study examines the impact of pH on a Tetrasphaera PAO enriched culture, over a pH range from 6.0 to 8.0 under both anaerobic and aerobic conditions, to assess its impact on the stoichiometry and kinetics of Tetrasphaera metabolism. It was discovered that the rates of phosphorus (P) uptake and P release increased with an increase of pH within the tested range, while PHA production, glycogen consumption and substrate uptake rate were less sensitive to pH changes. The results suggest that Tetrasphaera PAOs display kinetic advantages at high pH levels, which is consistent with what has been observed previously for Accumulibacter PAOs. The results of this study show that pH has a substantial impact on the P release and uptake kinetics of PAOs, where the P release rate was >3 times higher and the P uptake rate was >2 times higher at pH 8.0 vs pH 6.0, respectively. Process operational strategies promoting both Tetrasphaera and Accumulibacter activity at high pH do not conflict with each other, but lead to a potentially synergistic impact that can benefit EBPR performance.

Polyhydroxyalkanoates production from ethanol- and lactate-rich fermentate of confectionary industry effluents.

Polyhydroxyalkanoate (PHA) production has been the focus of considerable research to increase productivities and reduce production costs. In this study, a fermented confectionary industry wastewater was used as feedstock for mixed microbial culture PHA production. The feedstock was dominated by lactate and ethanol (60-90 % of all soluble fermentation products). The culture selection reactor was inoculated with municipal activated sludge and was operated at an organic loading rate (OLR) of 100 Cmmol·L-1·d-1, achieving a robust PHA-accumulating enrichment, which produced up to 52.6 ± 0.4 wt% of PHA in accumulation assays. An OLR increase in the culture selection stage to 150 Cmmol·L-1·d-1 led to a PHA content of 59.1 ± 0.6, a yield of 0.93 ± 0.01 Cmol-PHA·Cmol-S-1 and a productivity of 0.93 ± 0.01 g-PHA L-1·h-1. A correlation analysis of the impact of ethanol concentrations from 3.19 to 20.3 Cmmol·L-1 in the reactor showed that ethanol inhibited PHA production rate and yield and the consumption of other carbon sources available. Microbial community analysis revealed the increase of Amaricoccus genus during the bioreactor operation time, a known PHA accumulator. The produced polymer was poly(3-hydroxybutyrate) with an average molecular weight of 4.3 × 105 Da and a polydispersity index of 1.88.

Microbial population analysis of nutrient removal-related organisms in membrane bioreactors.

Membrane bioreactors (MBR) are an important and increasingly implemented wastewater treatment technology, which are operated at low food to microorganism ratios (F/M) and retain slow-growing organisms. Enhanced biological phosphorus removal (EBPR)-related organisms grow slower than ordinary heterotrophs, but have never been studied in detail in MBRs. This study presents a comprehensive analysis of the microorganisms involved in EBPR in pilot- and full-scale MBRs, using fluorescence in situ hybridization (FISH), as well as an overall assessment of other relevant microbial groups. The results showed that polyphosphate accumulating organisms (PAOs) were present at similar levels in all studied MBRs (10% ± 6%), even those without a defined anaerobic zone. Glycogen accumulating organisms were also detected, although rarely. The FISH results correlated well with the observed P removal performance of each plant. The results from this study suggest that a defined anaerobic zone is not necessarily required for putative PAO growth in MBRs, since polyphosphate storage may provide a selective advantage in fulfilling cell maintenance requirements in substrate-limited conditions (low F/M).

The impact of biomass withdrawal strategy on the biomass selection and polyhydroxyalkanoates accumulation of mixed microbial cultures.

The production of polyhydroxyalkanoates (PHA) by mixed microbial cultures (MMC) has been studied as an alternative to pure cultures in order to reduce the price of PHA through use of open systems and low-cost substrates, such as agro-industrial sub-products. However, the widespread applicability of this process depends on the optimization of operational factors impacting PHA productivity. This study addresses the impact of biomass withdrawal strategy on the performance of MMC selection reactors and consequently on biomass productivity and global PHA productivity. Two selection reactors were operated in parallel under similar conditions, except for the timing of biomass withdrawal, at the end of the famine phase (Reactor 1, R1) versus at the end of the feast phase (Reactor 2, R2) at an organic loading rate of 100 Cmmol.L-1.d-1 and solids retention time of 4 days. The biomass selected in both conditions had similar PHA storing capacity as shown by the similar yields of PHA per substrate obtained in the accumulation assays; however, R1 reached a higher biomass productivity (about 4-fold higher than R2). This study demonstrated that removing the excess biomass at the end of the famine phase resulted in a much higher global PHA productivity and that the key parameter affecting the global PHA productivity of the 2-stage system was the volumetric biomass productivity. Results obtained provide important insight into how MMC systems can be best operated to maximize PHA productivity.

Dynamics of Microbial Communities in Phototrophic Polyhydroxyalkanoate Accumulating Cultures.

Phototrophic mixed cultures (PMC) are versatile systems which can be applied for waste streams, valorisation and production of added-value compounds, such as polyhydroxyalkanoates (PHA). This work evaluates the influence of different operational conditions on the bacterial communities reported in PMC systems with PHA production capabilities. Eleven PMCs, fed either with acetate or fermented wastewater, and selected under either feast and famine (FF) or permanent feast (PF) regimes, were evaluated. Overall, results identified Chromatiaceae members as the main phototrophic PHA producers, along with Rhodopseudomonas, Rhodobacter and Rhizobium. The findings show that Chromatiaceae were favoured under operating conditions with high carbon concentrations, and particularly under the PF regime. In FF systems fed with fermented wastewater, the results indicate that increasing the organic loading rate enriches for Rhodopseudomonas, Rhizobium and Hyphomicrobiaceae, which together with Rhodobacter and Chromatiaceae, were likely responsible for PHA storage. In addition, high-sugar feedstock impairs PHA production under PF conditions (fermentative bacteria dominance), which does not occur under FF. This characterization of the communities responsible for PHA accumulation helps to define improved operational strategies for PHA production with PMC.

The impact of a seasonal change in loading rate on the nitrous oxide emissions at the WWTP of a tourist region.

Nitrous oxide (N2O) is a powerful greenhouse gas (GHG) whose production and emission must be minimised from wastewater treatment plants (WWTPs) to avoid undesirable impacts to climate change and the ozone layer. WWTPs operated in tourist regions undergo large seasonal changes to the influent loading rates of organic matter, nitrogen and phosphorus, which operators must respond to by changing their operational conditions. This study examines the impact of a change in low to high season on the N2O emissions of an activated sludge WWTP in a well-known tourist region in the Algarve, Portugal. While literature studies have suggested that increases in the nitrogen and organic loading rates can promote increased N2O emissions, we have found higher N2O emissions in the low season (7.4% kgN2O-N·kgNH4-N-1), where these loading rates were lower. It was found that the impact of accompanying operational changes to the WWTP outweighed any change caused by the increased loading rate, where the aeration rate showed a significant correlation with N2O emission dynamics. The location of the N2O fluxes observed as well as the dissolved vs gaseous N2O levels suggested that the hydroxylamine oxidation pathway was likely to be of higher relevance towards N2O production as compared to nitrifier denitrification. This study contributes towards the understanding of operational factors impacting N2O emissions at full-scale WWTPs and potential mitigation strategies.

Expanding ASM models towards integrated processes for short-cut nitrogen removal and bioplastic recovery.

In next-generation water resource recovery facilities (WRRFs), it is becoming increasingly important to save energy costs and promote resource recovery of valuable products. One way of reducing the substantial aeration energy costs at WRRFs is to employ shortcut N removal, while polyhydroxyalkanoate (PHA) production and recovery as bioplastic is a promising means of recovering a valuable product from biosolids. Both objectives can be achieved simultaneously through the Short-Cut Enhanced Phosphorus and PHA Recovery (SCEPPHAR) process. However, current mathematical models have not previously been employed to describe the behavior of such a process, which limits engineering design and optimisation of process operation. This work focusses on extending the ASM3 model towards the description of short-cut nitrogen removal and simultaneous PHA recovery in a sidestream treatment process. The calibrated and validated model described very well the nitritation process coupled with the aerobic feast/anoxic famine process for the selection of PHA producing organisms at a pilot-scale facility operated in Carbonera, Italy, where the normalised root mean squared error (NRMSE) was consistently <20%. Furthermore, the model applied to the PHA selection stage could effectively describe the PHA accumulation stage without recalibration. A simulation study was performed using the modified ASM3 model to assess the relative benefits of the SCEPPHAR process strategy as compared to the fully aerobic selection process for mixed culture PHA production. While the level of PHA production was found to be 34% lower with SCEPPHAR, a 43% savings in volatile fatty acids (VFAs) demand, a 15% decrease in Total suspended solids (TSS) production and a 28% decrease in oxygen demand were also achievable, which could lead to savings in operational costs. This study facilitates the design and optimisation of WRRFs that integrate short-cut N removal with PHA production, saving aeration energy costs while achieving resource recovery.

Diclofenac biotransformation in the enhanced biological phosphorus removal process.

Diclofenac is a pharmaceutical active compound frequently detected in wastewater and water bodies, and often reported to be persistent and difficult to biodegrade. While many previous studies have focussed on assessing diclofenac biodegradation in nitrification and denitrification processes, this study focusses on diclofenac biodegradation in the enhanced biological phosphorus removal (EBPR) process, where the efficiency of this process for diclofenac biodegradation as well as the metabolites generated are not well understood. An enrichment of Accumulibacter polyphosphate accumulating organisms (PAOs) was operated in an SBR for over 300 d, and acclimatized to 20 µg/L of diclofenac, which is in a similar range to that observed in domestic wastewater influents. The diclofenac biotransformation was monitored in four periods of stable operation and linked to the microbial community and metabolic behaviour in each period. Nitrification was observed in two of the four periods despite the addition of a nitrification inhibitor, and these periods were positively correlated with increased diclofenac biodegradation. Interestingly, in two periods with excellent phosphorus removal (>99%) and no nitrification, different levels of diclofenac biotransformation were observed. Period 2, enriched in Accumulibacter Type II achieved more significant diclofenac biotransformation (3.4 µg/gX), while period 4, enriched in Accumulibacter Type I achieved lower diclofenac biotransformation (0.4 µg/gX). In total, 23 transformation products were identified, with lower toxicity than the parent compound, enabling the elucidation of multiple metabolic pathways for diclofenac biotransformation. This study showed that PAOs can contribute to diclofenac biotransformation, yielding less toxic transformation products, and can complement the biodegradation carried out by other organisms in activated sludge, particularly nitrifiers.

A review of the biotransformations of priority pharmaceuticals in biological wastewater treatment processes.

Wastewater effluent discharges have been considered as one of the main sources of synthetic chemicals entering into the aquatic environment. Even though they occur at low concentrations, pharmaceutically active compounds (PhACs) can have an impact on ecological toxicity that affects aquatic organisms. Moreover, new regulations in development toward preserving water quality reinforces the increasing need to monitor and abate some PhACs in wastewater treatment plants (WWTPs), where they are typically only partially eliminated. Unlike most previous reviews, we have focussed on how the main biological and chemical molecular factors impact the biotransformations of key PhACs in biological WWTP processes. Biotransformations have been found to be an important contributor towards the removal of PhACs from WWTP effluents. This review paper critically assesses these aspects and the recent advances that have been achieved in wastewater treatment processes for biodegradation of 7 PhACs; namely the non-steroidal anti-inflammatory drug (NSAID) diclofenac (DCF); the macrolide antibiotics azithromycin (AZM), erythromycin (ERY) and clarithromycin (CLR); the two natural estrogens estrone (E1) and 17ß-estradiol (E2), and the synthetic estrogen 17α-ethinylesradiol (EE2). These represent the micropollutants of the EU Watch list in Decision 2015/495/EU that are most relevant to WWTPs due to their frequent detection. The metabolic pathways, transformation products and impact of relevant factors to biological WWTP processes is addressed in this review. The biokinetics of PhAC biodegradation in different engineered bioprocesses is also discussed. Promising technologies and operational strategies that are likely to have a high impact on controlling PhAC releases are highlighted and future research needs are also proposed.