Biotransformation Of Organic Micropollutants Research Articles

Deciphering the role of extracellular polymeric substances in the adsorption and biotransformation of organic micropollutants during anaerobic wastewater treatment

Extracellular polymeric substances (EPS) participate in the removal of organic micropollutants (OMPs), but the primary pathways of removal and detailed mechanisms remain elusive. We evaluated the effect of EPS on removal for 16 distinct chemical classes of OMPs during anaerobic digestion (AD). The results showed that hydrophobic OMPs (HBOMPs) could not be removed by EPS, while hydrophilic OMPs (HLOMPs) were amenable to removal via adsorption and biotransformation of EPS. The adsorption and biotransformation of HLOMPs by EPS accounted up to 19.4 ± 0.9 % and 6.0 ± 0.8 % of total removal, respectively. Further investigations into the adsorption and biotransformation mechanisms of HLOMPs by EPS were conducted utilizing spectral, molecular dynamics simulation, and electrochemical analysis. The results suggested that EPS provided abundant binding sites for the adsorption of HLOMPs. The binding of HLOMPs to tryptophan-like proteins in EPS formed nonfluorescent complexes. Hydrogen bonds, hydrophobic interactions and water bridges were key to the binding processes and helped stabilize the complexes. The biotransformation of HLOMPs by EPS may be attributed to the presence of extracellular redox active components (c-type cytochromes (c-Cyts), c-Cyts-bound flavins). This study enhanced the comprehension for the role of EPS on the OMPs removal in anaerobic wastewater treatment.

Quantitative structure-biotransformation relationships of organic micropollutants in aerobic and anaerobic wastewater treatments

Biotransformation is one of the dominant processes to remove organic micropollutants (OMPs) in wastewater treatment. However, studies on the role of molecular structure in determining the biotransformation rates of OMPs are limited. We evaluated the biotransformation of 14 OMPs belonging to different chemical classes under aerobic and anaerobic conditions, and then explored the quantitative structure-biotransformation relationships (QSBRs) of the OMPs based on biotransformation rates using valid molecular structure descriptors (electrical and physicochemical parameters). Pseudo-first-order kinetic modeling was used to fit the biotransformation rate, and only 2 of the 14 OMPs showed that the biotransformation rate constant (kbio) values were higher under anaerobic conditions than aerobic conditions, indicating that aerobic conditions were more favorable for biotransformation of most OMPs. QSBRs infer that the electrophilicity index (ω) is a reliable predictor for OMPs biotransformation under aerobic conditions. ω corresponds to the interaction between OMPs and microbial enzyme active sites, this process is the rate-limiting step of biotransformation. However, under anaerobic conditions the QSBR based on ω was not significant, indicating that specific functional groups may be more critical than electrophilicity. In conclusion, QSBRs can serve as alternative tools for the prediction of the biotransformation of OMPs and provide further insights into the factors that influence biotransformation.

Influence of metabolism and microbiology on organic micropollutants biotransformation in anoxic heterotrophic reactors.

There is scarce information about the biotransformation of organic micropollutants (OMPs) under anoxic conditions. In this study, a heterotrophic denitrifying bioreactor was set up to study the fate of several OMPs from metabolic and microbiological points of view. Primary metabolic activity was increased by adding progressively higher nitrogen loading rates during the operation (from 0.075 to 0.4gN-NO3- L-1 d-1), which resulted in an important shift in the microbial population from a specialized biomass to a more diverse community. Such a change provoked a significant increase in the removal efficiency of erythromycin (ERY), roxithromycin (ROX) and bisphenol-A (BPA), and some bacterial taxa, such as Rhodoplanes, were identified as possible indicators related to the biodegradation of these compounds. The increasing primary metabolic activity in the reactor did not enhance the OMP-specific removal rates, suggesting that the bacterial composition is more influential than cometabolism.

Biochar enhances the biotransformation of organic micropollutants (OMPs) in an anaerobic membrane bioreactor treating sewage

The removal of emerging organic micropollutants (OMPs) in anaerobic membrane bioreactors (AnMBRs) has garnered considerable attention owing to the rapid development of AnMBR technology and the increased environmental risk caused by OMP discharge. We investigated the removal efficiency of 22 typical OMPs from sewage being treated in an AnMBR, and implemented and evaluated an upgrading strategy by adding biochar. The average removal efficiency of OMPs was only 76.8% due to hydrophilic OMPs containing electron-withdrawing groups (ketoprofen, ibuprofen, diclofenac, and carbamazepine) being poorly removed. Biochar addition (5.0 g/L) promoted the removal of recalcitrant OMPs by 45%, leading to an enhanced removal efficiency of 88.7%. Although biochar has a high adsorption capacity to different OMPs, the biotransformation rather than sorption removal efficiency of 13 of the 22 OMPs was largely enhanced, suggesting that adsorption-biotransformation was the main approach by which biochar enhances the OMP removal. Biotransformation test and microbial analysis revealed that the enrichment of species (Flavobacterium, Massilia, Acinetobacter, and Cloacibacterium) involved in OMP biotransformation on biochar contributed largely to the enhanced biotransformation removal efficiency of OMPs. In this way, the enhanced electron transfer activity and syntrophic metabolism between hydrogenotrophic methanogens and species that oxidize acetate to H2/CO2 on biochar jointly contributed to the stable CH4 production and OMP biotransformation. This study provides a promising strategy to enhance the OMP removal in AnMBRs and improves our understanding of the underlying mechanism of biochar-amended OMP removal in anaerobic treatment systems.

Do NH4+-N and AOB affect atenolol removal during simulated riverbank filtration?

Biodegradation is regarding as the most important organic micro-pollutants (OMPs) removal mechanism during riverbank filtration (RBF), but the OMPs co-metabolism mechanism and the role of NH4+-N during this process are not well understood. Here, we selected atenolol as a typical OMP to explore the effect of NH4+-N concentration on atenolol removal and the role of ammonia oxidizing bacteria (AOB) in atenolol biodegradation. The results showed that RBF is an effective barrier for atenolol mainly by biodegradation and adsorption. The ratio of biodegradation and adsorption to atenolol removal was dependent on atenolol concentration. Specifically, atenolol with low concentration (500 ng/L) is almost completely removed by adsorption, while atenolol with higher concentration (100 μg/L) is removed by biodegradation (51.7%) and adsorption (30.8%). Long-term difference in influent NH4+-N concentrations did not show significant impact on atenolol (500 ng/L) removal, which was mainly dominated by adsorption. Besides, AOB enhanced the removal of atenolol (100 μg/L) as biodegradation played a more crucial role in removing atenolol under this concentration. Both AOB and heterotrophic bacteria can degrade atenolol during RBF, but the degree of AOB's contribution may be related to the concentration of atenolol exposure. The main reactions occurred during atenolol biodegradation possibly includes primary amide hydrolysis, hydroxylation and secondary amine depropylation. About 90% of the bio-transformed atenolol was produced as atenolol acid. AOB could transform atenolol to atenolol acid by inducing primary amide hydrolysis but failed to degrade atenolol acid further under the conditions of this paper. This study provides novel insights regarding the roles played by AOB in OMPs biotransformation during RBF.

Microaeration improves the removal/biotransformation of organic micropollutants in anaerobic wastewater treatment systems

This work assessed the effect of increasing microaeration flow rates (1–6 mL min−1 at 28 °C and 1 atm, equivalent to 0.025–0.152 L O2 L−1 feed) on the removal/biotransformation of seven organic micropollutants (OMPs) (three hormones, one xenoestrogen, and three pharmaceuticals), at 200 μg L−1 each, in a lab-scale upflow anaerobic sludge blanket reactor operated at a hydraulic retention time (HRT) of 7.4 h. Additionally, the operational stability of the system and the evolution of its microbial community under microaerobic conditions were evaluated. Microaeration was demonstrated to be an effective strategy to improve the limited removal/biotransformation of the evaluated OMPs in short-HRT anaerobic wastewater treatment systems. The rise in the airflow rate considerably increased the removal efficiencies of all OMPs. However, there seems to be a saturation limit for the biochemical reactions. Then, the best results were obtained with 4 mL air min−1 (0.101 L O2 L−1 feed) (~90%) because, above this flow rate, the efficiency increase was negligible. The long-term exposure to microaerobic conditions (249 days) led the microbiota to a gradual evolution. Consequently, there was some enrichment with species potentially associated with the biotransformation of OMPs, which may explain the better performance at the end of the microaerobic term even with the lowest airflow rate tested.

Heterotrophic enzymatic biotransformations of organic micropollutants in activated sludge.

While heterotrophic microorganisms constitute the major fraction of activated sludge biomass, the role of heterotrophs in the biotransformation of organic micropollutants (OMPs) has not been fully elucidated. Yet, such knowledge is essential, particularly when conceiving novel wastewater treatment plants based on a two-stage process including an A-stage under heterotrophic conditions and a B-stage based on anammox activity. Biotransformation of OMPs in activated sludge is thought to mostly occur cometabolically thanks to the action of low specificity enzymes involved in the metabolism of the primary substrates. For a better understanding of the process, it is important to determine such enzymatic activities and the underlying mechanisms involved in OMPs biotransformation. This task has proven to be difficult due to the lack of information about the enzymatic processes and the complexity of the biological systems present in activated sludge. In this paper, a continuous aerobic heterotrophic reactor following 20 OMPs at environmental concentrations was operated to (i) assess the potential of heterotrophs during the cometabolic biotransformation of OMPs, (ii) identify biotransformation reactions catalyzed by aerobic heterotrophs and (iii) predict possible heterotrophic enzymatic activities responsible for such biotransformations. Contradicting previous reports on the dominant role of nitrifiers in OMPs removal during activated sludge treatment, the heterotrophic population proved its capacity to biotransform the OMPs to extents equivalent to reported values in nitrifying activated sludge plants. Besides, 12 transformation products potentially formed through the activity of several enzymes present in heterotrophs, including monooxygenases, dioxygenases, hydrolases and transferases, were identified.

Role of Ammonia Oxidation in Organic Micropollutant Transformation during Wastewater Treatment: Insights from Molecular, Cellular, and Community Level Observations.

Organic micropollutants (OMPs) are a threat to aquatic environments, and wastewater treatment plants may act as a source or a barrier of OMPs entering the environment. Understanding the fate of OMPs in wastewater treatment processes is needed to establish efficient OMP removal strategies. Enhanced OMP biotransformation has been documented during biological nitrogen removal and has been attributed to the cometabolic activity of ammonia-oxidizing bacteria (AOB) and, specifically, to the ammonia monooxygenase (AMO) enzyme. Yet, the exact mechanisms of OMP biotransformation are often unknown. This critical review aims to fundamentally and quantitatively evaluate the role of ammonia oxidation in OMP biotransformation during wastewater treatment processes. OMPs can be transformed by AOB via direct and indirect enzymatic reactions: AMO directly transforms OMPs primarily via hydroxylation, while biologically produced reactive nitrogen species (hydroxylamine (NH2OH), nitrite (NO2-), and nitric oxide (NO)) can chemically transform OMPs through nitration, hydroxylation, and deamination and can contribute significantly to the observed OMP transformations. OMPs containing alkyl, aliphatic hydroxyl, ether, and sulfide functional groups as well as substituted aromatic rings and aromatic primary amines can be biotransformed by AMO, while OMPs containing alkyl groups, phenols, secondary amines, and aromatic primary amines can undergo abiotic transformations mediated by reactive nitrogen species. Higher OMP biotransformation efficiencies and rates are obtained in AOB-dominant microbial communities, especially in autotrophic reactors performing nitrification or nitritation, than in non-AOB-dominant microbial communities. The biotransformations of OMPs in wastewater treatment systems can often be linked to ammonium (NH4+) removal following two central lines of evidence: (i) Similar transformation products (i.e., hydroxylated, nitrated, and desaminated TPs) are detected in wastewater treatment systems as in AOB pure cultures. (ii) Consistency in OMP biotransformation (rbio, μmol/g VSS/d) to NH4+ removal (rNH4+, mol/g VSS/d) rate ratios (rbio/rNH4+) is observed for individual OMPs across different systems with similar rNH4+ and AOB abundances. In this review, we conclude that AOB are the main drivers of OMP biotransformation during wastewater treatment processes. The importance of biologically driven abiotic OMP transformation is quantitatively assessed, and functional groups susceptible to transformations by AMO and reactive nitrogen species are systematically classified. This critical review will improve the prediction of OMP transformation and facilitate the design of efficient OMP removal strategies during wastewater treatment.

The organic loading rate affects organic micropollutants’ cometabolic biotransformation kinetics under heterotrophic conditions in activated sludge

Several studies have shown that organic micropollutants (OMPs) are biotransformed cometabolically in activated sludge systems. However, the individual role of heterotrophs in the microbial consortium is still not clear, i.e., there is still a gap regarding the influence of the heterotrophic activity on the cometabolic biotransformation kinetics and yield of the OMPs. Aiming to answer these questions, experiments with increasing primary substrate concentrations were performed under aerobic heterotrophic conditions in a continuous stirred tank reactor operated at several organic loading rates (OLR) with fixed hydraulic retention time. Moreover, the individual kinetic parameters were determined in batch assays with different initial substrate concentrations using the sludges from the continuous reactor. A set of 15 OMPs displaying a variety of physicochemical properties were spiked to the feeding in the ng L−1 - µg L−1 range. Results reveal that the biodegradation of the primary carbon source and the biotransformation of the OMPs occur simultaneously, in clear evidence of cometabolic behavior. Moreover, we conclude that the OMPs biotransformation kinetic constant (kbiol) shows a linear dependence with the OLR of the primary substrate for most of the compounds studied, suggesting that the heterotrophic activity seriously affects the OMPs biotransformation kinetics. However, under typical activated sludge systems operating conditions (hydraulic retention times above 8 h), their biotransformation yield would not be significantly affected.

Acidogenesis is a key step in the anaerobic biotransformation of organic micropollutants

Understanding the role of the different anaerobic digestion stages on the removal of organic micropollutants (OMPs) is essential to mitigate their release from wastewater treatment plants. This study assessed the fate of 21 OMPs during hydrolysis and acidogenesis to elucidate the contribution of these stages to the overall anaerobic removal. Moreover, the removal mechanisms and factors influencing them were investigated. To this purpose, a fermentation reactor was operated and fed with two different substrates: starch (to jointly evaluate hydrolysis and acidogenesis) and glucose (to isolate acidogenesis). Results indicate that sulfamethoxazole was highly biotransformed (>80 %), while galaxolide, celestolide, tonalide, erythromycin, roxithromycin, trimethoprim, octylphenol and nonylphenol achieved a 50–80 % biotransformation. Since no significant differences in the biotransformation efficiencies were found between starch and glucose fermentation, it is stated that the enzymatic activities involved in starch hydrolysis do not significantly contribute to the cometabolic biotransformation of OMPs, while acidogenesis appears as the major player. Moreover, a higher biotransformation (≥15 percentage points and p ≤ 0.05) was found for galaxolide, celestolide, tonalide, erythromycin and roxithromycin during acidogenesis in comparison with the efficiencies reported for the acetogenic/methanogenic step. The biotransformation of some OMPs was explained considering their chemical structure and the enzymatic activities.

Thermal hydrolysis of sewage sludge partially removes organic micropollutants but does not enhance their anaerobic biotransformation

Pretreatment technologies prior to anaerobic digestion (AD) have been developed with the aim of enhancing biogas productivity and reducing the presence of pathogens in digested sludge. Among them, thermal hydrolysis (TH) appears as the most promising one. In wastewater treatment plants (WWTPs) sludge is the end point of many organic micropollutants (OMPs), which was proved to lead to important environmental and human risks since sludge is commonly used in agriculture. The objective of this work is to determine the fate OMPs in TH and subsequent AD. Sewage sludge was pretreated in a TH pilot plant at 170 °C for 20 min. Afterwards, two anaerobic digesters with a working volume of 14 L fed with fresh and pretreated sludge were operated in parallel in mesophilic conditions. TH proved to be an effective technology to partially or totally remove the dissolved fraction of OMPs as well as the fraction sorbed into those suspended solids that are solubilised after this pretreatment. However, it did not affect the OMPs sorbed concentration into solids that are not solubilised. Globally, the OMPs removal efficiency during TH appears to be linked to the solids solubilisation during this process. Afterwards, the OMPs biotransformation efficiency in AD of fresh and pretreated sludge was determined. Noticeable differences between the microbiome of both reactors was determined, but the anaerobic biotransformation was not substantially different for most of the OMPs. However, it affected musk fragrances, which presented considerably lower biotransformation efficiency in the reactor fed with pretreated sludge. Therefore, TH was proved effective in partially removing OMPs but not in enhancing their bioavailability and subsequent anaerobic biotransformation.

Biotransformation of organic micropollutants by anaerobic sludge enzymes

Biotransformation of organic micropollutants (OMPs) in wastewater treatment plants ultimately depends on the enzymatic activities developed in each biological process. However, few research efforts have been made to clarify and identify the role of enzymes on the removal of OMPs, which is an essential knowledge to determine the biotransformation potential of treatment technologies. Therefore, the purpose of the present study was to investigate the enzymatic transformation of 35 OMPs under anaerobic conditions, which have been even less studied than aerobic systems. Initially, 13 OMPs were identified to be significantly biotransformed (>20%) by anaerobic sludge obtained from a full-scale anaerobic digester, predestining them as potential targets of anaerobic enzymes. Native enzymes were extracted from this anaerobic sludge to perform transformation assays with the OMPs. In addition, the effect of detergents to recover membrane enzymes, as well as the effects of cofactors and inhibitors to promote and suppress specific enzymatic activities were evaluated. In total, it was possible to recover enzymatic activities towards 10 out of these 13 target OMPs (acetyl-sulfamethoxazole and its transformation product sulfamethoxazole, acetaminophen, atenolol, clarithromycin, citalopram, climbazole, erythromycin, and terbutryn, venlafaxine) as well as towards 8 non-target OMPs (diclofenac, iopamidol, acyclovir, acesulfame, and 4 different hydroxylated metabolites of carbamazepine). Some enzymatic activities likely involved in the anaerobic biotransformation of these OMPs were identified. Thereby, this study is a starting point to unravel the still enigmatic biotransformation of OMPs in wastewater treatment systems.

An innovative wastewater treatment technology based on UASB and IFAS for cost-efficient macro and micropollutant removal

An innovative process based on the combination of a UASB reactor and an IFAS system is proposed in order to combine different redox conditions and biomass conformations to promote a high microbial diversity. The objective of this configuration is to enhance the biological removal of organic micropollutants (OMPs) as well as to achieve the abatement of nitrogen by using the dissolved methane as an inexpensive electron donor. Results showed high removals of COD (93%) and dissolved methane present in the UASB effluent (up to 85%) was biodegraded by a consortium of aerobic methanotrophs and heterotrophic denitrifiers. Total nitrogen removal decreased slightly along the operation (from 44 to 33%), depending on the availability of electron donor, biomass concentration, and configuration (floccules and biofilm). A high removal was achieved in the hybrid system (>80%) for 6 of the studied OMPs. Sulfamethoxazole, trimethoprim, naproxen, and estradiol were readily biotransformed under anaerobic conditions, whereas ibuprofen or bisphenol A were removed in the anoxic-aerobic compartment. Evidence of the cometabolic biotransformation of OMPs has been found, such as the influence of nitrification activity on the removal of bisphenol A, and of the denitrification activity on ethinylestradiol removal.

A new decentralized biological treatment process based on activated carbon targeting organic micropollutant removal from hospital wastewaters

Although hospital wastewaters (HWWs) are usually discharged in urban sewage systems, their separate treatment has several benefits, such as the specific treatment of potential toxics as well as avoidance of further dilutions. In this work, an integrated industrial pilot plant (2200L) corresponding to the technology SeMPAC® is proposed and validated for such purpose. The process consists of a sequential batch reactor (SBR) connected to an external submerged microfiltration membrane, in which powdered activated carbon (PAC) is directly added into the biological reactor to enhance the removal of the organic micropollutants (OMPs). The combination of different redox conditions in the SBR, as well as the operation at long sludge retention times (SRTs) and high biomass concentrations favored OMP biotransformation in the SBR, being their final removal efficiencies enhanced clearly after PAC addition, especially for the recalcitrant compounds. A periodical renewal of the adsorbent is necessary to overcome its gradual saturation. The main operational conditions were influenced by (i) the recalcitrant OMP carbamazepine, which defines the PAC dosage; (ii) the easily degradable OMP ibuprofen, which can be used to optimize the duration of the aerobic cycle; and (iii) the denitrification efficiency, which defines the correct time length of the anoxic period.

Why are organic micropollutants not fully biotransformed? A mechanistic modelling approach to anaerobic systems

Biotransformation of most organic micropollutants (OMPs) during wastewater treatment is not complete and an unexplained steady decrease of the biotransformation rate with time is reported for many OMPs in different biological processes. To minimize and accurately predict the emission of OMPs into the environment, the mechanisms and limitations behind their biotransformations should be clarified. Aiming to achieve this objective, the present study follows a mechanistic modelling approach, based on the formulation of four models according to different biotransformation hypotheses: Michaelis-Menten kinetics, chemical equilibrium between the parent compound and the transformation product (TP), enzymatic inhibition by the TP, and a limited compound bioavailability due to its sequestration in the solid phase. These models were calibrated and validated with kinetic experiments performed in two different anaerobic systems: continuous reactors enriched with methanogenic biomass and batch assays with anaerobic sludge. Model selection was conducted according to model suitability criteria (goodness of fitting the experimental data, confidence of the estimated parameters, and model parsimony) but also considering mechanistic evidences. The findings suggest that reversibility of the biological reactions and/or sequestration of compounds are likely the causes preventing the complete biotransformation of OMPs, and biotransformation is probably limited by thermodynamics rather than by kinetics. Taking into account its simplicity and broader applicability spectrum, the reversible biotransformation is the proposed model to explain the incomplete biotransformation of OMPs.

Role of methanogenesis on the biotransformation of organic micropollutants during anaerobic digestion.

Several studies showed that some organic micropollutants (OMPs) are biotransformed during anaerobic digestion (AD). Yet, most of them aim at reporting removal efficiencies instead of understanding the biotransformation process. Indeed, how each of the main AD stages (i.e., hydrolysis, acidogenesis, and methanogenesis) contribute to OMP biotransformation remains unknown. This study focuses on investigating the role of methanogenesis, the most characteristic step of AD, to OMP removal. More specifically, the sorption and the biotransformation of 20 OMPs by methanogenic biomass were analyzed determining their concentrations in both liquid and solid phases. Sorption onto methanogenic biomass displayed a similar behavior as reported for digested sludge. Most of the OMPs were biotransformed to a medium extent (35-70%) and only sulfamethoxazole was completely removed. Comparing these results with those reported for the complete AD process, methanogenesis was proven to play a key role, accounting for more than 50% of the OMP biotransformation (except for roxithromycin) during AD. An increase in the organic loading rate from 1 to 2gCOD/Ld, typical loads employed in sewage sludge anaerobic digesters, did not exert a clear cometabolic effect on the OMPs biotransformation. It is hypothesized that biotransformation occurs in both liquid and solid phases because no link between the partition coefficient (Kd) and the overall biotransformation efficiency was found. These findings allow a better understanding of the OMPs fate under anaerobic conditions, which is necessary to design efficient biological mitigation strategies.

Cometabolic Enzymatic Transformation of Organic Micropollutants under Methanogenic Conditions.

Anaerobic digestion (AD) has been shown to have the biological potential to decrease concentrations of several organic micropollutants (OMPs) in sewage sludge. However, the mechanisms and factors behind these biotransformations, which are essential for elucidating the possible transformation products and to foster the complete removal of OMPs via operational strategies, remain unclear. Therefore, this study investigated the transformation mechanisms of 20 OMPs during the methanogenic step of AD with a focus on the role of acetate kinase (AK), which is a key enzyme in methane production. The results from lab-scale methanogenic reactors showed that this step accounts for much of the reported OMP biotransformation in AD. Furthermore, enzymatic assays confirmed that AK transforms galaxolide, naproxen, nonylphenol, octylphenol, ibuprofen, diclofenac, bisphenol A, and triclosan. Except for galaxolide, for which further studies are required to refine conclusions, the OMP's chemical structure was a determinant for AK action because only compounds that contain a carboxyl or hydroxyl group and have moderate steric hindrance were enzymatically transformed, likely by phosphorylation. For these seven compounds, this enzymatic mechanism accounts for 10-90% of the measured methanogenic biotransformation, suggesting that other active enzymes of the AD process are also involved in OMP biotransformation.