Diagnosing Down-the-Drain Disposal of Unused Pharmaceuticals at a River Catchment Level: Unrecognized Sources of Environmental Contamination That Require Nontechnological Solutions.

Down-the-drain disposal of pharmaceuticals remains an overlooked and unrecognized source of environmental contamination that requires nontechnological "at-source" solutions. Monitoring of 31 pharmaceuticals over 7 days in five wastewater treatment plants (WWTPs) serving five cities in South-West UK revealed down-the-drain codisposal of six pharmaceuticals to three WWTPs (carbamazepine and propranolol in city A, sildenafil in city B, and diltiazem, capecitabine, and sertraline in city D), with a one-off record codisposal of estimated 253 pills = 40 g of carbamazepine and estimated 96 pills = 4 g of propranolol in city A accounting for their 10- and 3-fold respective increases in wastewater daily loads. Direct disposal of pharmaceuticals was found to affect the efficiency of wastewater treatment with much higher pharmaceutical removal (decrease in daily load) during "down-the-drain disposal" days. This is due to lack of conjugated glucuronide metabolites that are cleaved during "consumption-only" days, with the release of a parent pharmaceutical counterbalancing its removal. Higher removal of pharmaceuticals during down-the-drain disposal days reduced pharmaceutical loads reaching receiving environment, albeit with significant levels remaining. The estimated daily loads in receiving water downstream from a discharge point accounted for 13.8 ± 3.4 and 2.1 ± 0.2 g day-1 of carbamazepine and propranolol, respectively, during consumption-only days and peaked at 20.9 g day-1 (carbamazepine) and 4.6 g day-1 (propranolol) during down-the-drain disposal days. Actions are needed to reduce down-the-drain disposal of pharmaceuticals. Our recent work indicated that down-the-drain disposal of pharmaceuticals doubled since the last study in 2005, which may be due to the lack of information and messaging that informs people to dispose of unused medicines at pharmacies. Media campaigns that inform the public of how to safely dispose of medicines are key to improving rates of return and reducing pharmaceutical waste in the environment. The environment is a key motivator for returning unused medicines to a pharmacy and so messaging should highlight environmental risks associated with improper disposal.

Multi-residue ultra-performance liquid chromatography coupled with tandem mass spectrometry method for comprehensive multi-class anthropogenic compounds of emerging concern analysis in a catchment-based exposure-driven study.

This paper presents a new multi-residue method for the quantification of more than 142 anthropogenic compounds of emerging concern (CECs) in various environmental matrices. These CECs are from a wide range of major classes including pharmaceuticals, household, industrial and agricultural. This method utilises ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS) for analysis of five matrices (three liquid and two solid) from wastewater treatment processes and the surrounding environment. Relative recoveries were predominantly between 80 and 120%; however, due to the complexity of the matrices used in this work, not all compounds were recovered in all matrices, from 138/142 analytes in surface water to 96/142 analytes in digested solids. Method quantification limits (MQLs) ranged from 0.004 ng L-1 (bisoprolol in surface water) to 3118 ng L-1 (creatinine in wastewater treatment work (WwTW) influent). The overall method accuracy was 107.0%, and precision was 13.4%. To test its performance, the method was applied to the range of environmental matrices at WwTWs in South West England. Overall, this method was found to be suitable for application in catchment-based exposure-driven studies, as, of the total number of analytes quantifiable in each matrix, 61% on average was found to be above their corresponding MQL. The results confirm the need for analysing both the liquid and solid compartments within a WwTW to prevent under-reporting of concentrations.

Assessing the impact of soil microbial fuel cells on atrazine removal in soil.

Widespread pesticide use in agriculture is a major source of soil pollution, driving biodiversity loss and posing serious threads to human health. The recalcitrant nature of most of these pesticides demands for effective remediation strategies. In this study, we assess the ability of soil microbial fuel cell (SMFC) technology to bioremediate soil polluted by the model pesticide atrazine. To elucidate the degradation mechanism and consequently define effective implementation strategies, we provide the first comprehensive investigation of the SMFC performance, in which the monitoring of the electrochemical performance of the system is combined with Quadrupole Time-of-Flight (QTOF) mass spectrometry and microbial analyses. Our results show that, while both SMFC and natural attenuation lead to a reduction on atrazine levels, the SMFC modulates the activity of different microbial pathways. As a result, atrazine degradation by natural attenuation leads to high levels of deisoproylatrazine (DIPA), a very toxic degradation metabolite, while DIPA levels in soil treated by SMFC remain comparatively low. The beta diversity and differential abundance analyses revealed how the microbial community evolves over time in the SMFCs degrading atrazine, demonstrating the enrichment of electroactive taxa on the anode, and the enrichment of a mixture of electroactive and atrazine-degrading taxa at the cathode. The detection and taxonomic classification of peripheral atrazine degrading genes, atzA, atzB and atzC, was carried out in combination with the differential abundance analysis. Results revealed that these genes are likely harboured by members of the order Rhizobiales enriched at the cathode, thus promoting atrazine degradation via the conversion of hydroxyatrazine (HA) into N-isopropylammelide (NIPA), as confirmed by mass spectrometry data. Overall, the comprehensive approach adopted in this work, provides fundamental insights into the degradation pathways of atrazine in soil by SMFC technology, which is critical for practical applications, thus suggesting an effective approach to advance research in the field.

Multi-biomarker approach for estimating population size in a national-scale wastewater-based epidemiology study.

This study identifies biochemical markers (BCIs) that can be used as population markers in wastewater-based epidemiology (WBE) and compares their estimates with other established population size estimation (PE) methods, including census data (PECEN). Several groups of BCIs (64 targets: genetic and chemical markers) were investigated in an intercity study, including 10 cities/towns within England equating to a population of ∼7 million people. Several selection criteria were applied to identify the best BCIs to provide robust estimation of population size at a catchment level: (1) excellent performance with analytical methods; (2) excellent fit of the linear regression model which indicates PE-driven BCI daily loads; (3) low temporal variability in usage; (4) human-linked origin. Only a few tested BCIs showed a strong positive linear correlation between daily BCI loads and PE indicating their low spatiotemporal variability. These are: cimetidine, clarithromycin, metformin, cotinine, bezafibrate, metronidazole and hydroxymetronidazole, diclofenac, and benzophenone 1. However, only high/long term usage pharmaceuticals: cimetidine and metformin as well as cotinine (metabolite of nicotine) performed well when tested in two independent datasets and catchments accounting for both spatial and temporal scales. Strong seasonal usage trends were observed for antihistamines, NSAIDs (anti-inflammatories), antibiotics and UV filters, invalidating them as PE markers. Key conclusions from the study are: (1) Cimetidine is the best performing BCI; (2) Chemical markers outperform genetic markers as PE BCIs; (3) Water utility PE estimates (PEWW) align well with PECEN and PEBCI values; (4) Ammonium/orthophosphate as well as viral PE markers suffer from high temporal variability, hence, they are not recommended as PEBCI markers, and, most importantly, (5) PEBCI calibration/validation at the country/region level is advised in order to establish the best PE markers suited for local/national needs and accounting for site/region specific uncertainties.

Ketogenic diet but not free-sugar restriction alters glucose tolerance, lipid metabolism, peripheral tissue phenotype, and gut microbiome: RCT.

Restricted sugar and ketogenic diets can alter energy balance/metabolism, but decreased energy intake may be compensated by reduced expenditure. In healthy adults, randomization to restricting free sugars or overall carbohydrates (ketogenic diet) for 12 weeks reduces fat mass without changing energy expenditure versus control. Free-sugar restriction minimally affects metabolism or gut microbiome but decreases low-density lipoprotein cholesterol (LDL-C). In contrast, a ketogenic diet decreases glucose tolerance, increases skeletal muscle PDK4, and reduces AMPK and GLUT4 levels. By week 4, the ketogenic diet reduces fasting glucose and increases apolipoprotein B, C-reactive protein, and postprandial glycerol concentrations. However, despite sustained ketosis, these effects are no longer apparent by week 12, when gut microbial beta diversity is altered, possibly reflective of longer-term adjustments to the ketogenic diet and/or energy balance. These data demonstrate that restricting free sugars or overall carbohydrates reduces energy intake without altering physical activity, but with divergent effects on glucose tolerance, lipoprotein profiles, and gut microbiome.

An integrated One Health framework for holistic evaluation of risks from antifungal agents in a large-scale multi-city study.

A new framework for retrospective mass spectral data mining for antifungal agents (AFs) and Wastewater-Based Epidemiology (WBE) was developed as part of One Health framework to tackle risks from AFs. A large scale, multi-city study was undertaken in South-West England. Key drivers of AFs in the catchment were identified with communal wastewater discharges being the main driver for human AFs (fluconazole, ketoconazole) and agricultural runoff being the main driver for pesticide AFs (prochloraz, prothioconazole and tebuconazole). Average WBE-estimated human used fluconazole and ketoconazole PNDIs (population normalised daily intake) exceeded 300 mg day-1 1000 inh-1 and 2000 mg day-1 1000 inh-1. This is much higher than PNDPs (population normalised daily prescriptions <40 mg day-1 1000 inh-1 and <80 mg day-1 1000 inh-1 for fluconazole and ketoconazole respectively). This was expected due to both prescription and over-the-counter usage, and both oral and topical applications. Pesticide AF, prothioconazole had PNDIs <40,000 mg day-1 1000 inh -1, which gave intake: 0.43, 0.26, 0.07 mg kg-1 in City A, B, and C, likely due to accounting for external/non-human sources. This is higher than the acceptable daily intake (ADI) of 0.01 mg kg-1bw day-1, which warrants further study. Intake per kg of body weight estimated using tebuconazole was 0.86, 1.39, 0.12, 0.13, and 2.7 mg kg-1 in City A-E respectively and is likely due to external/non-human sources. Intake calculated using its metabolite was 0.02 and 0.01 mg kg-1 in City B and C respectively, which aligned with ADI (0.03 mg kg-1bw day-1). The environmental risk assessment of AFs indicated low/medium risk from fluconazole, prochloraz, and tebuconazole, medium risk from epoxiconazole, prothioconazole's metabolite, and tebuconazole, and high risk for prothioconazole in river water. High risk was estimated from fluconazole, epoxiconazole, prothioconazole and its metabolite, tebuconazole, ketoconazole in wastewater samples, which is important during raw sewage discharge events via sewer overflows.

Wastewater-based epidemiology for comprehensive community health diagnostics in a national surveillance study: Mining biochemical markers in wastewater.

This manuscript showcases results from a large scale and comprehensive wastewater-based epidemiology (WBE) study focussed on multi-biomarker suite analysis of both chemical and biological determinants in 10 cities and towns across England equating to a population of ∼7 million people. Multi-biomarker suite analysis, describing city metabolism, can provide a holistic understanding to encompass all of human, and human-derived, activities of a city in a single model: from lifestyle choices (e.g. caffeine intake, nicotine) through to health status (e.g. prevalence of pathogenic organisms, usage of pharmaceuticals as proxy for non-communicable disease, NCD, conditions or infectious disease status), and exposure to harmful chemicals due to environmental and industrial sources (e.g. pesticide intake via contaminated food and industrial exposure). Population normalised daily loads (PNDLs) of many chemical markers were found, to a large extent, driven by the size of population contributing to wastewater (especially NCDs). However, there are several exceptions providing insights into chemical intake that can inform either disease status in various communities or unintentional exposure to hazardous chemicals: e.g. very high PNDLs of ibuprofen in Hull resulting from its direct disposal (confirmed by ibuprofen/2-hydroxyibuprofen ratios) and bisphenol A (BPA) in Hull, Lancaster and Portsmouth likely related to industrial discharge. An importance for tracking endogenous health markers such as 4-hydroxy-2-nonenal-mercapturic acid (HNE-MA, an oxidative stress marker) as a generic marker of health status in communities was observed due to increased levels of HNE-MA seen at Barnoldswick wastewater treatment plant that coincided with higher-than-average paracetamol usage and SARS-CoV-2 prevalence in this community. PNDLs of virus markers were found to be highly variable. Being very prevalent in communities nationwide during sampling, SARS-CoV-2 presence in wastewater was to a large extent community driven. The same applies to the fecal marker virus, crAssphage, which is very prevalent in urban communities. In contrast, norovirus and enterovirus showed much higher variability in prevalence across all sites investigated, with clear cases of localized outbreaks in some cities while maintaining low prevalence in other locations. In conclusion, this study clearly demonstrates the potential for WBE to provide an integrated assessment of community health which can help target and validate policy interventions aimed at improving public health and wellbeing.

Human population as a key driver of biochemical burden in an inter-city system: Implications for One Health concept.

This paper tests the hypothesis that human population and city function are key drivers of biochemical burden in an inter-city system, which can be used to inform One Health actions as it enables a holistic understanding of city's metabolism encompassing all of the activities of a city in a single model: from lifestyle choices, through to health status and exposure to harmful chemicals as well as effectiveness of implemented management strategies. Chemical mining of wastewater for biophysico-chemical indicators (BCIs) was undertaken to understand speciation of BCIs in the context of geographical as well as community-wide socioeconomic factors. Spatiotemporal variabilities in chemical and biological target groups in the studied inter-city system were observed. A linear relationship (R2 > 0.99) and a strong positive correlation between most BCIs and population size (r > 0.998, p < 0.001) were observed which provides a strong evidence for the population size as a driver of BCI burden. BCI groups that are strongly correlated with population size and are intrinsic to humans' function include mostly high usage pharmaceuticals that are linked with long term non-communicable conditions (NSAIDs, analgesics, cardiovascular, mental health and antiepileptics) and lifestyle chemicals. These BCIs can be used as population size markers. BCIs groups that are produced as a result of a specific city's function (e.g. industry presence and occupational exposure or agriculture) and as such are not correlated with population size include: pesticides, PCPs and industrial chemicals. These BCIs can be used to assess city's function, such as occupational exposure, environmental or food exposure, and as a proxy of community-wide health. This study confirmed a strong positive correlation between antibiotics (ABs), population size and antibiotic resistance genes (ARGs). This confirms the population size and AB usage as the main driver of AB and ARG levels and provides an opportunity for interventions aimed at the reduction of AB usage to reduce AMR. Holistic evaluation of biophysicochemical fingerprints (BCI burden) of the environment and data triangulation with socioeconomic fingerprints (indices) of tested communities are required to fully embrace One Health concept.

Occurrence of pharmaceutical residues, personal care products, lifestyle chemicals, illicit drugs and metabolites in wastewater and receiving surface waters of Krakow agglomeration in South Poland.

This is the first study of broad range of chemical classes CECs conducted in the upper Wisla river catchment including the biggest WWTPs in this region and surface waters. The list of compounds is extensive and the paper provides, for the first time, better understanding of environmental burden from PCPCs in Poland. Cumulative contribution of hypertension pharmaceuticals, nonsteroidal anti-inflammatory drugs (NSAIDs) and lifestyle chemicals was 89% and 95% in wastewater influent, and 75% in wastewater effluent at both WWTPs. Significant removal efficiencies, exceeding 90%, were found for parabens, UV filters, NSAIDs, steroid estrogens, plasticizers, antibacterials/antibiotics, stimulants and metabolites and lifestyle chemicals. The comparison of the average mass loads of CECs between the influent and effluent, has shown that 27% and 29% of all detected CECs were removed by less than 50%. An increase of concentrations of CECs in the effluent was observed for 18% and 20% of all detected CECs in Kujawy and Plaszow WWTPs, respectively. Negative mass balances of fexofenadine, venlafaxine, o-desmethyltramadol, ketamine and temazepam were noted within WWTPs, which are a result of dissolution of persistent contaminants accumulated in aggregates and/or back-transformation or de-conjugation of metabolites into parent compounds. 44 CECs were detected in surface waters located upstream and downstream of the WWTPs. The concentrations of compounds were largely dependent on the dilution factor of WWTP discharge. The risk quotation (RQ) values for compounds present in surface waters were calculated in relation to their potential for bioaccumulation. Among compounds with high potential for bioaccumulation, with log KOW ≥ 4.5, diclofenac, atorvastatin and triclosan were found to be of high risk. Many CECs with high, moderate or even low environmental impact have shown high potential for bioaccumulation and should be considered as priority at the same risk level. Moreover, possible synergistic action is still of concern.

Spatiotemporal profiling of antibiotics and resistance genes in a river catchment: Human population as the main driver of antibiotic and antibiotic resistance gene presence in the environment.

Studies to understand the role wastewater treatment plants (WWTPs) play in the dissemination of antibiotics (ABs), and in the emergence of antibiotic resistance (ABR), play an important role in tackling this global crisis. Here we describe the abundance and distribution of 16 ABs, and 4 corresponding antibiotic resistance genes (ARGs), sampled from the influent to five WWTPs within a single river catchment. We consider four classes of antibiotics: fluroquinolones, macrolides, sulfamethoxazole and chloramphenicol, as well the corresponding antibiotic resistance genes qnrS, ermB, sul1 and catA. All antibiotics, apart from four fluroquinolones (besifloxacin, lomefloxacin, ulifloxacin, prulifloxacin), were detected within all influent wastewater from the 5 cities (1 city = 1 WWTP), as were the corresponding antibiotic resistance genes (ARGs). Strong correlations were observed between the daily loads of ABs and ARGs versus the size of the population served by each WWTP, as well as between AB and ARG loads at a single site. The efficiency of ABs and ARGs removal by the WWTPs varied according to site (and treatment process utilized) and target, although strong correlations were maintained between the population size served by WWTPs and daily loads of discharged ABs and ARGs into the environment. We therefore conclude that population size is the main determinant of the magnitude of AB and ARG burden in the environment.

Micropollutant fluxes in urban environment - A catchment perspective.

This study provided a holistic understanding of the sources, fate and behaviour of 142 compounds of emerging concern (CECs) throughout a river catchment impacted by 5 major urban areas. Of the incoming 169.3 kg d-1 of CECs entering the WwTWs, 167.9 kg d-1 were present in the liquid phase of influent and 1.4 kg d-1 were present in the solid phase (solid particulate matter, SPM). Analysis of SPM was important to determine accurate loads of incoming antidepressants and antifungal compounds, which are primarily found in the solid phase. Furthermore, these classes and the plasticiser, bisphenol A (BPA) were the highest contributors to CEC load in digested solids. Population normalised loads showed little variation across the catchment at 154 ± 12 mg d-1 inhabitant-1 indicating that population size is the main driver of CECs in the studied catchment. Across the catchment 154.6 kg d-1 were removed from the liquid phase during treatment processes. CECs discharged into surface waters from individual WwTWs contributed between 0.19 kg d-1 at WwTW A to 7.3 kg d-1 at WwTW E, which correlated strongly with the respective contributing populations. Spatial and temporal variations of individual CECs and their respective classes were found in WwTW influent (both solid (influentSPM) and liquid phases (influentAQ)) throughout the catchment, showing that different urban areas impact the catchment in different ways, with key variables being lifestyle, use of over-the-counter pharmaceuticals and industrial activity. Understanding of both spatial and temporal variation of CECs at the catchment level helped to identify possible instances of direct disposal, as in the case of carbamazepine. Analysis of surface waters throughout the catchment showed increasing mass loads of CECs from upstream of WwTW A to downstream at WwTW D, showing clear individual contributions from WwTWs. Many CECs were ubiquitous throughout the river water in the catchment. Daily loads ranged from 0.005 g d-1 (ketamine, WwTW A) up to 1890.3 g d-1 (metformin, WwTW C) for the 84/138 CECs that were detected downstream of the WwTWs. For metformin this represents the equivalent of ∼1,890 tablets (1,000 mg per tablet) dissolved in the river water downstream of WwTW C.

Estimation of community-wide multi-chemical exposure via water-based chemical mining: Key research gaps drawn from a comprehensive multi-biomarker multi-city dataset.

This paper explores the strong potential of chemical mining of wastewater for markers of community-wide intake of wide-ranging harmful chemicals belonging to several usage groups: industrial chemicals, personal care products, pesticides, illicit drugs, lifestyle chemicals and prescription pharmaceuticals as a proxy for multi-chemical community-wide exposure. An estimation of chemical intake in five contrasting town/cities based in the Avon River catchment in the South-West UK was undertaken. High-resolution spatiotemporal pharmaceutical prescription databases were used for system calibration, both in terms of biomarker selection and its correction factor, as well as for the overall system performance evaluation, both spatially and temporality. Only metabolism data accounting for phase two metabolism provided correct estimates of pharma intake. Using parent compounds as XCRs (xenobiotic compounds residue) was found to overestimate exposure due to an inclusion of directly disposed (unused) drugs. Spatiotemporal trends in XC intake were observed as a result of occupational exposure (higher bisphenol A (BPA) intake during weekday), and lifestyle choices (higher cocaine and pyrethroid pesticides intake during weekend). WBE is not intended to estimate individual exposure to chemicals. It can however provide estimates at a community level, and as a result, it has the potential to be developed into an early warning system, a powerful tool for large scale screening studies identifying communities at risk and in need of high resolution individual testing at a localised scale.

(Fluoro)quinolones and quinolone resistance genes in the aquatic environment: A river catchment perspective.

This study provides an insight into the prevalence of (fluoro)quinolones (FQs) and their specific quinolone qnrS resistance gene in the Avon river catchment area receiving treated wastewater from 5 wastewater treatment plants (WWTPs), serving 1.5 million people and accounting for 75% of inhabitants living in the catchment area in the South West of England.. Ofloxacin, ciprofloxacin, nalidixic acid and norfloxacin were found to be ubiquitous with daily loads reaching a few hundred g/day in wastewater influent and tens of g/day in receiving waters. This was in contrast to other FQs analysed: flumequine, nadifloxacin, lomefloxacin, ulifloxacin, prulifloxacin, besifloxacin and moxifloxacin, which were hardly quantified. Enantiomeric profiling revealed that ofloxacin was enriched with the S-(-)-enantiomer, likely deriving from its prescription as the more potent enantiomerically pure levofloxacin, alongside racemic ofloxacin. While ofloxacin's enantiomeric fraction (EF) remained constant, high stereoselectivity was observed in the case of its metabolite ofloxacin-N-oxide. The removal efficiency of quinolones during wastewater treatment at 5 WWTPs utilising either trickling filters (TF) or activated sludge (AS), was compound and wastewater treatment process dependent, with AS providing better efficiency than TF. The qnrS resistance gene was ubiquitous in wastewater. Its removal was WWTP treatment process dependent with TF performing best and resulting in significant removal of the gene (from 28 to 75%). AS underperformed with only 9% removal in the case of activated sludge and actual increase in the gene copy number within sequencing batch reactors (SBRs). Interestingly, the data suggests that higher removal of antibiotics could be linked with high prevalence of the gene (SBR and WWTP E) and vice versa, low removal of antibiotic is correlated with lower prevalence of the gene in wastewater effluent (TF, WWTP B and D). This is especially prominent in the case of ofloxacin and could indicate that AS might be facilitating antimicrobial resistance (AMR) prevalence to higher extent than TF. Wastewater-based epidemiology (WBE) was also applied to monitor any potential misuse (e.g. direct disposal) of FQs in the catchment. In most cases higher use of antibiotics with respect to official statistics (i.e. ciprofloxacin, ofloxacin) was observed, which suggests that FQs management practice require further attention.

Estimation of community-wide exposure to bisphenol A via water fingerprinting.

Molecular epidemiology in human biomonitoring allows for verification of public exposure to chemical substances. Unfortunately, due to logistical difficulties and high cost, it evaluates only small study groups and as a result does not provide comprehensive large scale community-wide exposure data. Wastewater fingerprinting utilizing metabolic biomarkers of exposure that are excreted collectively by studied populations into urine and ultimately into the community's wastewater, provides a timely alternative to traditional approaches. This study aimed to provide comprehensive spatiotemporal community-wide exposure to bisphenol A (BPA, including BPA intake) using wastewater fingerprinting. Wastewater fingerprinting was undertaken using high resolution mass spectrometry retrospective data mining of characteristic BPA human metabolism marker (bisphenol A sulphate), applied to a large geographical area of 2000 km2 and a population of ~1.5 million served by 5 WWTPs (wastewater treatment plants) accounting for >75% of the overall population in the studied catchment. Community-wide BPA intake was found to be below temporary tolerable daily intake (t-TDI) level of 4 µg kg-1 day-1 set by the European Food Safety Agency (EFSA) suggesting overall low exposure at 3 WWTPs serving residential areas with low industrial/commercial presence. However, at two WWTPs serving communities with higher industrial/commercial presence, higher BPA sulphate loads corresponding to higher (up to 14 times) BPA intakes (exceeding 10 µg kg-1 day-1 at one WWTP and reaching 50 µg kg-1 day-1 at the second WWTP) were observed and they are likely linked with occupational exposure. Characteristic temporal variations of BPA intake were noted in most studied WWTPs with the lowest intake occurring during weekends and the highest during weekdays.

Assessment of bisphenol-A in the urban water cycle.

The plasticizer bisphenol-A (BPA) is common to municipal wastewaters and can exert toxicity to exposed organisms in the environment. Here BPA concentration at 5 sewage treatment works (STW) and distribution throughout a river catchment in South West UK were investigated. Sampling sites included influent and effluent wastewater (n = 5), river water (n = 7) and digested sludge (n = 2) which were monitored for 7 consecutive days. Findings revealed average BPA loads in influent wastewater at two STWs were 10-37 times greater than the other wastewaters monitored. Concentrations up to ~100 µg L-1 were measured considerably higher than previously reported for municipal wastewaters. Temporal variability throughout the week (i.e., highest concentrations during weekdays) suggests these high concentrations are linked with industrial activity. Despite ≥90% removal during wastewater treatment, notable concentrations remained in tested effluent (62-892 ng L-1). However, minimal impact on BPA concentrations in river water was observed for any of the effluents. The maximum BPA concentration found in river water was 117 ng L-1 which is considerably lower than the current predicted no effect concentration of 1.6 µg L-1. Nevertheless, analysis of digested sludge from sites which received these elevated BPA levels revealed average concentrations of 4.6 ±â€¯0.3 and 38.7 ±â€¯5.4 µg g-1. These sludge BPA concentrations are considerably greater than previously reported and are attributed to the high BPA loading in influent wastewater. A typical sludge application regime to agricultural land would result in a predicted BPA concentration of 297 ng g-1 in soil. Further studies are needed on the toxicological thresholds of exposed terrestrial organisms in amended soils to better assess the environmental risk here.

Stereoisomeric profiling of chiral pharmaceutically active compounds in wastewaters and the receiving environment - A catchment-scale and a laboratory study.

Chiral pharmaceutically active compounds (cPACs) are not currently governed by environmental regulation yet are expected to be in the future. As cPACs can exert stereospecific toxicity in the aquatic environment, it is essential to better understand their stereoselective behaviour here. Therefore, this study aims to provide a new perspective towards comprehensive evaluation of cPACs at a river catchment level, including their stereochemistry as a chemical phenomenon driving fate of chiral molecules in the environment. A large spatial and temporal monitoring program was performed in Southwest England. It included 5 sewage treatment works and the receiving waters of the largest river catchment in Southwest England. Simultaneously, lab-scale microcosm studies in simulated activated sludge bioreactors and river water microcosm were performed to evaluate stereoselective degradation of cPACs. A multi-residue enantioselective method allowed the analysis of a total of 18 pairs of enantiomers and 3 single enantiomers in wastewater and river water samples. Our monitoring program revealed: (1) spatial and temporal variations of cPACs in influent wastewaters resulting from different patterns of usage as well as an (2) enantiomeric enrichment of cPACs, likely due to human metabolism, despite their commercialization as racemic mixtures. A similar chiral signature was observed in effluent and receiving waters. Stereoselective degradation was observed in trickling filters (TF) for naproxen, ketoprofen, cetirizine and 10,11-dihydroxy-10-hydroxycarbamazepine, in sequencing batch reactors (SBR) for ifosfamide and in activated sludge (AS) for cetirizine. The extent of enantiomer-specific fate was wastewater treatment dependent in the case of naproxen (TF showed higher stereoselectivity than AS and SBR) and cetirizine (TF and AS showed higher stereoselectivity than SBR) due to differing microbial population. Furthermore, stereoselective degradation of naproxen was highly variable among STWs using similar treatments (TF) and operating in the same region. Microbial stereoselective degradation was also confirmed by both activated and river water simulated microcosm for chloramphenicol, ketoprofen, indoprofen, naproxen and 10,11-dihydroxy-10-hydroxycarbamazepine. Results from our large scale river catchment monitoring study and lab simulated microcosm show wide-ranging implications of enantiomerism of cPACs on environmental risk assessment (ERA). As two enantiomers of the same compound show different biological effects (e.g. toxicity), their non-racemic presence in the environment might lead to inaccurate ERA. This is because current ERA approaches do not require analysis at enantiomeric level.

Stereochemistry of ephedrine and its environmental significance: Exposure and effects directed approach.

Analysis of drugs and pharmaceuticals in the environment is typically performed with non-chiral chromatographic techniques. The environmental risks posed by chiral compounds analysed in this way must therefore be assumed to be independent of chirality, meaning that each enantiomer is equally potent in toxicity and long-lived in stability. This manuscript examines the degradation of each of the four isomers of ephedrine in river simulating microcosms and links this to toxicity data obtained by exposing three different organisms (D. magna, P. subcapitata and T. thermophila) to each of the isomers individually. Microcosms showed that significant degradation only occurred in biotic conditions and that only two isomers (1R,2S-(-)-ephedrine, 1S,2S-(+)-pseudoephedrine) degraded significantly over a period of fourteen days. This is concerning because at least one of the non-degraded isomers (1S,2R-(+)-ephedrine) has been observed in wastewater effluent, which discharges directly into rivers, meaning these isomers could be persistent in the environment. We also observed formation of 1S,2R-(+)-ephedrine in single isomer 1R,2S-(-)-ephedrine river simulating microcosms. Human liver microsome assays and mass spectrometry based data mining revealed that 1S,2R-(+)-ephedrine is not human derived but it could be formed as a results of microbial metabolic processes. Across all three organisms tested the persistent isomers (1S,2R-(+)-ephedrine and 1R,2R-(-)-pseudoephedrine) were more toxic than those that undergo degradation; meaning that if these isomers are entering or formed in the environment they might represent a potentially hazardous contaminant.

Biotic phase micropollutant distribution in horizontal sub-surface flow constructed wetlands.

The distribution of micropollutants in biotic phases of horizontal sub-surface flow (HSSF) constructed wetlands was investigated. 88 diverse micropollutants (personal care products, pharmaceuticals and illicit drugs) were monitored for in full-scale HSSF steel slag and gravel beds to assess their fate and behaviour during tertiary wastewater treatment. Of the studied micropollutants 54 were found in receiving and treated wastewaters. Treatment reduced concentrations of several micropollutants by >50% (removal range -112% to 98%) and resulted in changes to the stereo-isomeric composition of chiral species. For example, stereo-selective changes were observed for 3,4-methylenedioxymethamphetamine (MDMA) and atenolol during HSSF constructed wetland treatment for the first time. Analysis of sludge present within the HSSF beds found 37 micropollutants to be present. However, concentrations for the majority of these micropollutants were not considered high enough to suggest partitioning into sludge was a contributing mechanism of removal. Nevertheless the preservative methylparaben was found at 2772mgbed-1. Its daily removal from wastewater of 3.4mgd-1 indicates partitioning and accumulation in sludge contributes to its removal. Other micropollutants found at high levels in sludge (relative to their overall removals) were the antidepressants sertraline and fluoxetine, and the metabolite desmethylcitalopram. Furthermore, process balances indicated uptake and metabolism by Phragmites australis (Cav.) Trin. ex Steud did not contribute significantly to micropollutant removal. However analysis of plant tissues evidenced uptake, metabolism and accumulation of recalcitrant micropollutants such as ketamine and carbamazepine. It is considered that the rate of uptake was too slow to have a notable impact on removal at the 14h hydraulic retention time. Despite evidence of other removal mechanisms at play (e.g., partitioning into sludge and plant uptake), findings indicate biodegradation is the dominant mechanism of micropollutant removal in HSSF constructed wetlands.

Critical evaluation of monitoring strategy for the multi-residue determination of 90 chiral and achiral micropollutants in effluent wastewater.

It is essential to monitor the release of organic micropollutants from wastewater treatment plants (WWTPs) for developing environmental risk assessment and assessing compliance with legislative regulation. In this study the impact of sampling strategy on the quantitative determination of micropollutants in effluent wastewater was investigated. An extended list of 90 chiral and achiral micropollutants representing a broad range of biological and physico-chemical properties were studied simultaneously for the first time. During composite sample collection micropollutants can degrade resulting in the under-estimation of concentration. Cooling collected sub-samples to 4°C stabilised ≥81 of 90 micropollutants to acceptable levels (±20% of the initial concentration) in the studied effluents. However, achieving stability for all micropollutants will require an integrated approach to sample collection (i.e., multi-bottle sampling with more than one stabilisation method applied). Full-scale monitoring of effluent revealed time-paced composites attained similar information to volume-paced composites (influent wastewater requires a sampling mode responsive to flow variation). The option of monitoring effluent using time-paced composite samplers is advantageous as not all WWTPs have flow controlled samplers or suitable sites for deploying portable flow meters. There has been little research to date on the impact of monitoring strategy on the determination of chiral micropollutants at the enantiomeric level. Variability in wastewater flow results in a dynamic hydraulic retention time within the WWTP (and upstream sewerage system). Despite chiral micropollutants being susceptible to stereo-selective degradation, no diurnal variability in their enantiomeric distribution was observed. However, unused medication can be directly disposed into the sewer network creating short-term (e.g., daily) changes to their enantiomeric distribution. As enantio-specific toxicity is observed in the environment, similar resolution of enantio-selective analysis to more routinely applied achiral methods is needed throughout the monitoring period for accurate risk assessment.

Enantioselective transformation of fluoxetine in water and its ecotoxicological relevance.

European legislation focusing on water quality is expected to broaden to encompass several pharmaceuticals as priority hazardous substances. This manuscript aims to challenge current regulatory approaches that do not recognize stereochemistry of chiral pharmaceuticals by testing the hypothesis that environmental transformation and effects of chiral pharmaceuticals are stereoselective. Our experiments revealed that, while degradation of chiral fluoxetine (FL) in river water occurs via non-enantioselective photochemical and mildly-enantioselective microbial processes favoring the (R)-enantiomer, a pronounced enantioselectivity favoring (S)-FL (leading to the formation of (S)-NFL (norfluoxetine)) is observed during activated sludge treatment. Toxicity tests proved strong enantiomer-specific toxicity in the case of Tetrahymena thermophila, protozoa that are utilized during activated sludge treatment ((R)-FL is 30× more toxic than (S)-FL; (S)-NFL is 10× more toxic than (S)-FL). This is of paramount importance as preferential degradation of (S)-FL in activated sludge microcosms leads to the enrichment of FL with 30× more toxic (R)-FL and formation of 10× more toxic (S)-NFL. It is commonly assumed that a decreased concentration of FL leads to decreased biological impact. Our study proves that despite the overall decrease in FL concentration, accumulation of toxic (R)-FL and formation of toxic (S)-NFL leads to much higher than presumed toxicological effects.