Antimicrobial Transformation Products in the Aquatic Environment: Global Occurrence, Ecotoxicological Risks, and Potential of Antibiotic Resistance.

The global spread of antimicrobial resistance (AMR) is concerning for the health of humans, animals, and the environment in a One Health perspective. Assessments of AMR and associated environmental hazards mostly focus on antimicrobial parent compounds, while largely overlooking their transformation products (TPs). This review lists antimicrobial TPs identified in surface water environments and examines their potential for AMR promotion, ecological risk, as well as human health and environmental hazards using in silico models. Our review also summarizes the key transformation compartments of TPs, related pathways for TPs reaching surface waters and methodologies for studying the fate of TPs. The 56 antimicrobial TPs covered by the review were prioritized via scoring and ranking of various risk and hazard parameters. Most data on occurrences to date have been reported in Europe, while little is known about antibiotic TPs in Africa, Central and South America, Asia, and Oceania. Occurrence data on antiviral TPs and other antibacterial TPs are even scarcer. We propose evaluation of structural similarity between parent compounds and TPs for TP risk assessment. We predicted a risk of AMR for 13 TPs, especially TPs of tetracyclines and macrolides. We estimated the ecotoxicological effect concentrations of TPs from the experimental effect data of the parent chemical for bacteria, algae and water fleas, scaled by potency differences predicted by quantitative structure-activity relationships (QSARs) for baseline toxicity and a scaling factor for structural similarity. Inclusion of TPs in mixtures with their parent increased the ecological risk quotient over the threshold of one for 7 of the 24 antimicrobials included in this analysis, while only one parent had a risk quotient above one. Thirteen TPs, from which 6 were macrolide TPs, posed a risk to at least one of the three tested species. There were 12/21 TPs identified that are likely to exhibit a similar or higher level of mutagenicity/carcinogenicity, respectively, than their parent compound, with tetracycline TPs often showing increased mutagenicity. Most TPs with increased carcinogenicity belonged to sulfonamides. Most of the TPs were predicted to be mobile but not bioaccumulative, and 14 were predicted to be persistent. The six highest-priority TPs originated from the tetracycline antibiotic family and antivirals. This review, and in particular our ranking of antimicrobial TPs of concern, can support authorities in planning related intervention strategies and source mitigation of antimicrobials toward a sustainable future.

A mood state-specific interaction between kynurenine metabolism and inflammation is present in bipolar disorder.

OBJECTIVES: Cytokines are thought to contribute to the pathogenesis of psychiatric symptoms by kynurenine pathway activation. Kynurenine metabolites affect neurotransmission and can cause neurotoxicity. We measured inflammatory markers in patients with bipolar disorder (BD) and studied their relation to kynurenine metabolites and mood. METHODS: Patients with BD suffering from an acute mood episode were assigned to the depressive (n = 35) or (hypo)manic (n = 32) subgroup. Plasma levels of inflammatory markers [cytokines, C-reactive protein] and kynurenine metabolites [tryptophan (TRP), kynurenine (KYN), 3-hydroxykynurenine (3-HK), quinolinic acid (QA), kynurenic acid (KYNA)] were measured on 6 time points during 8 months follow-up. Biological marker levels in patients were compared to controls (n = 35) and correlated to scores on mood scales. Spearman correlations and linear mixed models were used for statistical analysis. RESULTS: Twenty patients of the manic subgroup, 29 of the depressive subgroup, and 30 controls completed the study. The manic subgroup had a rapid remission of mood symptoms, but in the depressive subgroup subsyndromal symptoms persisted. No differences in inflammation were found between groups. A strong correlation between tumor necrosis factor-α and KYN, KYN/TRP, 3-HK and QA (ρ > 0.60) was specific for the manic group, but only at baseline (during mania). The depressive subgroup had a lower neuroprotective ratio (KYNA/3-HK, P = .0004) and a strong association between interferon-y and kynurenine pathway activation (P < .0001). KYNA was low in both patient groups versus controls throughout the whole follow-up (P = .0008). CONCLUSIONS: Mania and chronic depressive symptoms in BD are accompanied by a strong interaction between inflammation and a potentially neurotoxic kynurenine metabolism.

Suspect and Nontargeted Strategies to Investigate in Vitro Human Biotransformation Products of Emerging Environmental Contaminants: The Benzotriazoles.

Benzotriazole derivatives (BTRs) are high production volume chemicals involved in a wide range of applications and consumer products resulting in their ubiquitous presence in environmental matrices. Yet, the human exposure assessment to these chemicals is limited since it is based only on the analysis of parent compounds in biological matrices. The objective of this study was to investigate the in vitro human biotransformation for three widely used BTRs and to stepwise examine the role of Phase I and II enzymes (cytochrome P450 (CYP), uridine glucuronic acid transferase (UGT), and sulfotransferase (SULT)) in their biotransformation. Extracts with generated biotransformation products (bioTPs) were analyzed using liquid chromatography coupled to quadrupole-time-of-flight mass spectrometry (LC-QTOF-MS), followed by their identification based on a workflow combining suspect and nontargeted strategies. Ten bioTPs were identified for 1H-benzotriazole, 14 for tolyltriazole, and 14 for 5-chloro-1H-benzotriazole. Most of the proposed bioTPs were identified and structurally elucidated for the first time. Based on these findings, possible bioTPs and metabolic transformation pathways were subsequently predicted for other structurally close BTR derivatives. Our findings provide new identified in vitro biotransformation products for future biomonitoring studies and emphasize that it is important to investigate the biotransformation pathway to assess overall exposure to xenobiotics.

Mining the Chemical Information on Urban Wastewater: Monitoring Human Exposure to Phosphorus Flame Retardants and Plasticizers.

At the individual level, exposure to contaminants is generally assessed through the analysis of specific biomarkers in biological matrices. However, these studies are costly and logistically demanding, limiting their applicability to monitor population-wide exposure over time and space. By focusing on a selection of exposure biomarkers to phosphorus flame retardants and plasticizers (PFRs), this study aims to explore the possibility of using wastewater as a complementary source of information about exposure. Wastewater samples were collected from five cities in Europe and analyzed using a previously established method. Substantial differences in biomarker levels were observed between the investigated catchments, suggesting differences in exposure. Time trends in biomarkers observed between 2013 and 2016 were found to agree with results from human biomonitoring studies and reports about production volumes. Using Monte Carlo simulations, average urinary concentrations were estimated. These were generally higher compared to results from human biomonitoring studies. Various explanations for these differences were formulated (i.e., other excretion routes, external sources and different sampling approaches). Obtained results show that wastewater analysis provides unique information about geographical and temporal differences in exposure, which would be difficult to gather using other monitoring tools.

Liquid Chromatography-Tandem Mass Spectrometry Analysis of Biomarkers of Exposure to Phosphorus Flame Retardants in Wastewater to Monitor Community-Wide Exposure.

Phosphorus flame retardants and plasticizers (PFRs) are increasingly used in consumer goods, from which they can leach and pose potential threats to human health. Monitoring human exposure to these compounds is thus highly relevant. Current assessment of exposure through analysis of biological matrices is, however, tedious as well as logistically and financially demanding. Analysis of selected biomarkers of exposure to PFRs in wastewater could be a simple and complementary approach to monitoring, over space and time, exposure at the population level. An analytical procedure, based on solid-phase extraction (SPE) and liquid chromatography coupled to tandem mass spectrometry, was developed and validated to monitor the occurrence in wastewater of human exposure biomarkers of 2-ethylhexyldiphenyl phosphate (EHDPHP), tris(2-butoxyethyl) phosphate (TBOEP), triphenyl phosphate (TPHP), tris(2-chloroisopropyl) phosphate (TCIPP), and tris(2-chloroethyl) phosphate (TCEP). Various SPE sorbents and extraction protocols were evaluated, and for the optimized method, absolute extraction recoveries ranged between 46% and 100%. Accuracy and precision were satisfactory for the selected compounds. Method detection limits ranged from 1.6 to 19 ng L-1. Biomarkers of exposure to PFRs were measured for the first time in influent wastewater. Concentrations in samples collected in Belgium ranged from below the limit of quantitation to 1072 ng L-1, with 2-ethylhexyl phenyl phosphate (EHPHP) and TCEP being the most abundant. Per capita loads of target biomarkers varied greatly, suggesting potential differences in exposure between the investigated communities. The developed method allowed implementation of the concepts of human biomonitoring at the community scale, opening the possibility to assess population-wide exposure to PFRs.

Novel Wastewater-Based Epidemiology Approach Based on Liquid Chromatography-Tandem Mass Spectrometry for Assessing Population Exposure to Tobacco-Specific Toxicants and Carcinogens.

Tobacco smoking remains an important public health issue worldwide. Assessment of exposure to tobacco-related toxicants and carcinogens at the population level is thus an essential population health indicator. This can be achieved by wastewater-based epidemiology (WBE), which relies on the analysis of biomarkers in wastewater. However, required analytical methods for the simultaneous measurement of tobacco-related toxicants and carcinogens in wastewater are not available. In this study, a new analytical procedure was developed and validated to measure tobacco-related alkaloids, carcinogens, and their metabolites in raw wastewater, including anabasine (ANABA), anatabine (ANATA), cotinine (COT), trans-3'-hydroxycotinine (COT-OH), N-nitrosoanabasine (NAB), N-nitrosoanatabine (NAT), N-nitrosonornicotine (NNN), 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), NNAL-N-ß-glucuronide, and NNAL-O-ß-glucuronide. Different parameters were optimized for the solid-phase extraction procedure and instrumental analysis using liquid chromatography-tandem mass spectrometry. The optimized method was fully validated, resulting in acceptable within-run and between-run precision (<8% and <10% relative standard deviation, respectively) and accuracy (<9% and <13% bias, respectively). Method quantification limits were at 0.5-120 ng/L in wastewater. Target analytes were stable in wastewater at 4 and 20 °C over 24 h. The developed method was applied to wastewater samples from two Belgian cities. Average concentrations of COT, COT-OH, ANATA, ANABA, and NAT were 5200, 2600, 30, 10, and 0.6 ng/L, respectively, while NAB, NNN, NNK, and NNAL were not detected in the samples. With the developed robust analytical method, our study provided the first insight into the population exposure to both toxicants and carcinogens resulting from tobacco use.

Can wastewater-based epidemiology be used to evaluate the health impact of temperature? - An exploratory study in an Australian population.

Ambient temperature is known to have impact on population health but assessing its impact by the traditional cohort approach is resource intensive. Wastewater-based epidemiology (WBE) could be an alternative for the traditional approach. This study was to provide the first evaluation to see if WBE can be used to assess the impact of temperature exposure to a population in South East Queensland, Australia using selected pharmaceuticals and personal care products (PPCPs) as biomarkers. Daily loads of eight PPCPs in wastewater collected from a wastewater treatment plant were measured from February 2011 to June 2012. Corresponding daily weather data were obtained from the closest weather station. Missing data of PPCPs were handled using the multiple imputation (MI) method, then we used a one-way between-groups analysis of variance to examine the seasonal effect on daily variation of PPCPs by seasons. Finally, an MI estimate was performed to evaluate the continuous relationship between daily average temperature and each multiply-imputed PPCP using time-series regression analysis. The results indicated that an increase of 1°C in average temperature associated with decrease at 1.3g/d (95% CI: -2.2 to (-0.4), p<0.05) for atenolol, increase at 36.5g/d (95% CI: 25.2-47.8, p<0.01) for acesulfame, and increase at 0.8g/d (95% CI: 0.02-1.55, p=0.05) for naproxen. No significant association was observed between temperature and the remaining PPCPs, comprising: caffeine, carbamazepine, codeine, hydrochlorothiazide, and salicylic acid. The findings suggested that consumption of sweetened drinks, risk of worsening cardiovascular conditions and pains are associated with variation in ambient temperature. WBE can thus be used as a complementary method to traditional cohort studies in epidemiological evaluation of the association between environmental factors and health outcomes provided that specific biomarkers of such health outcomes can be identified.

Evaluation of Monitoring Schemes for Wastewater-Based Epidemiology to Identify Drug Use Trends Using Cocaine, Methamphetamine, MDMA and Methadone.

Wastewater-based epidemiology is increasingly being used as a tool to monitor drug use trends. To minimize costs, studies have typically monitored a small number of days. However, cycles of drug use may display weekly and seasonal trends that affect the accuracy of monthly or annual drug use estimates based on a limited number of samples. This study aimed to rationalize sampling methods for minimizing the number of samples required while maximizing information about temporal trends. A range of sampling strategies were examined: (i) targeted days (e.g., weekends), (ii) completely random or stratified random sampling, and (iii) a number of sampling strategies informed by known weekly cycles in drug use data. Using a time-series approach, analysis was performed for four drugs (MDMA, methamphetamine, cocaine, methadone) collected through a continuous sampling program over 14 months. Results showed, for drugs with weekly cycles (MDMA, methamphetamine and cocaine in this sample), sampling strategies which made use of those weekly cycles required fewer samples to obtain similar information as sampling 5 days per week and had better accuracy than stratified random sampling techniques.

Systematic and day-to-day effects of chemical-derived population estimates on wastewater-based drug epidemiology.

Population size is crucial when estimating population-normalized drug consumption (PNDC) from wastewater-based drug epidemiology (WBDE). Three conceptually different population estimates can be used: de jure (common census, residence), de facto (all persons within a sewer catchment), and chemical loads (contributors to the sampled wastewater). De facto and chemical loads will be the same where all households contribute to a central sewer system without wastewater loss. This study explored the feasibility of determining a de facto population and its effect on estimating PNDC in an urban community over an extended period. Drugs and other chemicals were analyzed in 311 daily composite wastewater samples. The daily estimated de facto population (using chemical loads) was on average 32% higher than the de jure population. Consequently, using the latter would systemically overestimate PNDC by 22%. However, the relative day-to-day pattern of drug consumption was similar regardless of the type of normalization as daily illicit drug loads appeared to vary substantially more than the population. Using chemical loads population, we objectively quantified the total methodological uncertainty of PNDC and reduced it by a factor of 2. Our study illustrated the potential benefits of using chemical loads population for obtaining more robust PNDC data in WBDE.

Could wastewater analysis be a useful tool for China? - A review.

Analysing wastewater samples is an innovative approach that overcomes many limitations of traditional surveys to identify and measure a range of chemicals that were consumed by or exposed to people living in a sewer catchment area. First conceptualised in 2001, much progress has been made to make wastewater analysis (WWA) a reliable and robust tool for measuring chemical consumption and/or exposure. At the moment, the most popular application of WWA, sometimes referred as sewage epidemiology, is to monitor the consumption of illicit drugs in communities around the globe, including China. The approach has been largely adopted by law enforcement agencies as a device to monitor the temporal and geographical patterns of drug consumption. In the future, the methodology can be extended to other chemicals including biomarkers of population health (e.g. environmental or oxidative stress biomarkers, lifestyle indicators or medications that are taken by different demographic groups) and pollutants that people are exposed to (e.g. polycyclic aromatic hydrocarbons, perfluorinated chemicals, and toxic pesticides). The extension of WWA to a huge range of chemicals may give rise to a field called sewage chemical-information mining (SCIM) with unexplored potentials. China has many densely populated cities with thousands of sewage treatment plants which are favourable for applying WWA/SCIM in order to help relevant authorities gather information about illicit drug consumption and population health status. However, there are some prerequisites and uncertainties of the methodology that should be addressed for SCIM to reach its full potential in China.

A model to estimate the population contributing to the wastewater using samples collected on census day.

An important uncertainty when estimating per capita consumption of, for example, illicit drugs by means of wastewater analysis (sometimes referred to as "sewage epidemiology") relates to the size and variability of the de facto population in the catchment of interest. In the absence of a day-specific direct population count any indirect surrogate model to estimate population size lacks a standard to assess associated uncertainties. Therefore, the objective of this study was to collect wastewater samples at a unique opportunity, that is, on a census day, as a basis for a model to estimate the number of people contributing to a given wastewater sample. Mass loads for a wide range of pharmaceuticals and personal care products were quantified in influents of ten sewage treatment plants (STP) serving populations ranging from approximately 3500 to 500 000 people. Separate linear models for population size were estimated with the mass loads of the different chemical as the explanatory variable: 14 chemicals showed good, linear relationships, with highest correlations for acesulfame and gabapentin. De facto population was then estimated through Bayesian inference, by updating the population size provided by STP staff (prior knowledge) with measured chemical mass loads. Cross validation showed that large populations can be estimated fairly accurately with a few chemical mass loads quantified from 24-h composite samples. In contrast, the prior knowledge for small population sizes cannot be improved substantially despite the information of multiple chemical mass loads. In the future, observations other than chemical mass loads may improve this deficit, since Bayesian inference allows including any kind of information relating to population size.

Monitoring substance use in prisons: Assessing the potential value of wastewater analysis.

Prison substance use is a major concern for prison authorities and the wider community. Australia has responded to this problem by implementing the National Corrections Drug Strategy. Across Australia, the true extent of prison substance use cannot be determined. As a result, the effectiveness of the interventions employed as part of this strategy cannot be properly assessed. This has important implications for the allocation of corrective services resources and future policy development. This article explores the benefits and limitations, as well as the ethical and practical issues in using wastewater analysis (WWA) to measure levels of substance use in prisons. It reports results from the first application of WWA to an Australian prison, which supports the use of WWA in this context. Given the increasing concern for prescription misuse in prisons, we also highlight the novel use of WWA to measure the extent of prescription misuse by prisoners. The article concludes that as a result of its objectivity, sensitivity and cost-effectiveness, the use of WWA in prisons warrants further consideration in Australia.

Are we using more sugar substitutes? Wastewater analysis reveals differences and rising trends in artificial sweetener usage in Swedish urban catchments.

The market for artificial sweeteners as substitutes for conventional sugar (sucrose) is growing, despite potential health risks associated with their intake. Estimating population usage of artificial sweeteners is therefore crucial, and wastewater analysis can serve as a complement to existing methods. This study evaluated spatial and temporal usage of artificial sweeteners in five Swedish communities based on wastewater analysis. We further compared their levels measured in wastewater with the restrictions during the COVID-19 pandemic in Sweden and assessed health risks to the Swedish population. Influent wastewater samples (n = 194) collected in March 2019-February 2022 from communities in central and southern Sweden were analyzed for acesulfame, saccharin, and sucralose using liquid-chromatography coupled with tandem mass spectrometry. Spatial differences in loads for individual artificial sweetener were observed, with sucralose being higher in Kalmar (southern Sweden), and acesulfame and saccharin in Enköping and Östhammar (central Sweden). Based on sucrose equivalent doses, all communities showed a consistent prevalence pattern of sucralose > acesulfame > saccharin. Four communities with relatively short monitoring periods showed no apparent temporal changes in usage, but the four-year monitoring in Uppsala revealed a significant (p < 0.05) annual increase of ∼19 % for sucralose, ∼9 % for acesulfame and ∼8 % for saccharin. This trend showed no instant or delayed effects from COVID-19 restrictions, reflecting positively on the studied population which retained similar exposure to the artificial sweeteners despite potential pandemic stresses. Among the three artificial sweeteners, only acesulfame's levels were at the lower end of the health-related threshold for consumption of artificially sweetened beverages; yet, all were far below the acceptable daily intake, indicating no appreciable health risks. Our study provided valuable, pilot insights into the spatio-temporal usage of artificial sweeteners in Sweden and their associated health risks. This shows the usefulness of wastewater analysis for public health authorities wishing to assess future relevant interventions.

Making waves: The NORMAN antibiotic resistant bacteria and resistance genes database (NORMAN ARB&ARG)-An invitation for collaboration to tackle antibiotic resistance.

With the global concerns on antibiotic resistance (AR) as a public health issue, it is pivotal to have data exchange platforms for studies on antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) in the environment. For this purpose, the NORMAN Association is hosting the NORMAN ARB&ARG database, which was developed within the European project ANSWER. The present article provides an overview on the database functionalities, the extraction and the contribution of data to the database. In this study, AR data from three studies from China and Nepal were extracted and imported into the NORMAN ARB&ARG in addition to the existing AR data from 11 studies (mainly European studies) on the database. This feasibility study demonstrates how the scientific community can share their data on AR to generate an international evidence base to inform AR mitigation strategies. The open and FAIR data are of high potential relevance for regulatory applications, including the development of emission limit values / environmental quality standards in relation to AR. The growth in sharing of data and analytical methods will foster collaboration on risk management of AR worldwide, and facilitate the harmonization in the effort for identification and surveillance of critical hotspots of AR. The NORMAN ARB&ARG database is publicly available at: https://www.norman-network.com/nds/bacteria/.

Workflow to facilitate the detection of new psychoactive substances and drugs of abuse in influent urban wastewater.

The complexity around the dynamic markets for new psychoactive substances (NPS) forces researchers to develop and apply innovative analytical strategies to detect and identify them in influent urban wastewater. In this work a comprehensive suspect screening workflow following liquid chromatography - high resolution mass spectrometry analysis was established utilising the open-source InSpectra data processing platform and the HighResNPS library. In total, 278 urban influent wastewater samples from 47 sites in 16 countries were collected to investigate the presence of NPS and other drugs of abuse. A total of 50 compounds were detected in samples from at least one site. Most compounds found were prescription drugs such as gabapentin (detection frequency 79%), codeine (40%) and pregabalin (15%). However, cocaine was the most found illicit drug (83%), in all countries where samples were collected apart from the Republic of Korea and China. Eight NPS were also identified with this protocol: 3-methylmethcathinone 11%), eutylone (6%), etizolam (2%), 3-chloromethcathinone (4%), mitragynine (6%), phenibut (2%), 25I-NBOH (2%) and trimethoxyamphetamine (2%). The latter three have not previously been reported in municipal wastewater samples. The workflow employed allowed the prioritisation of features to be further investigated, reducing processing time and gaining in confidence in their identification.

Phytoextraction of per- and polyfluoroalkyl substances (PFAS) and the influence of supplements on the performance of short-rotation crops.

Per- and polyfluoroalkyl substances (PFAS) are anthropogenic compounds threatening water quality and food safety worldwide. Phytoremediation is a nature-based, cost-effective, and scalable solution with high potential for treating PFAS-contaminated sites. However, there is a large knowledge gap regarding choice of plant species and methods to enhance performance. This study assessed the PFAS phytoextraction potential of sunflower (Helianthus annuus), mustard (Brassica juncea), and industrial hemp (Cannabis sativa) in a greenhouse experiment, using inorganic fertilizer and a microbial mixture as supplements. PFAS concentrations were measured using UPLC-MS/MS, and bioconcentration factors for different plant tissues and removal efficiency were determined. Perfluoroalkyl carboxylic acid (PFCA) accumulation was 0.4-360 times higher than that of perfluoroalkyl sulfonic acid (PFSA) homologues of similar perfluorocarbon chain length. Inorganic fertilizer significantly (p < 0.001) reduced PFAS concentration in all plant tissues, whereas the microbial mixture tested did not affect PFAS concentration. PFAS uptake ranged from 0.2 to 33% per crop cycle. Overall, the potential number of crop cycles required for removal of 90% of individual PFAS ranged from six (PFPeA) to 232 (PFOA) using sunflower, 15 (PFPeA) to 466 (PFOS) using mustard and nine (PFPeA) to 420 (PFOS) using Hemp. In this study, the percentage of PFAS removal by plants was determined, and an estimation of the time required for PFAS phytoextraction was determined for the first time. This information is important for practical phytoremediation applications.

Degradation of 75 organic micropollutants in fresh human urine and water by UV advanced oxidation process.

In household wastewater, a large proportion of organic micropollutants (OMPs) load is attributed to human urine. OMPs could pose a risk to human and environmental health when urine collected in source-separating sanitation systems is recycled as crop fertiliser. This study evaluated degradation of 75 OMPs in human urine treated by a UV-based advanced oxidation process. Fresh urine and water samples were spiked with a broad range of OMPs and fed into a photoreactor equipped with a UV lamp (185 and 254 nm) that generated free radicals in situ. Degradation rate constant and the energy required to degrade 90% of all the OMPs in both matrices were determined. At a UV dose of 2060 J m-2, average ΣOMP degradation of 99% (±4%) in water and 55% (±36%) in fresh urine was achieved. The energy demand for removal of OMPs in water was <1500 J m-2, but for removal of OMPs in urine at least 10-fold more energy was needed. A combination of photolysis and photo-oxidation can explain the degradation of OMPs during UV treatment. Organic substances (e.g. urea, creatinine) likely inhibited degradation of OMPs in urine by competitively absorbing UV-light and scavenging free radicals. There was no reduction in the nitrogen content of urine during treatment. In summary, UV treatment can reduce the load of OMPs to urine recycling sanitation systems.

Impact of a megacity on the water quality of a tropical estuary assessed by a combination of chemical analysis and in-vitro bioassays.

Tropical estuaries are threatened by rapid urbanization, which leads to the spread of thousands of micropollutants and poses an environmental risk to such sensitive aqueous ecosystems. In the present study, a combination of chemical and bioanalytical water characterization was applied to investigate the impact of Ho Chi Minh megacity (HCMC, 9.2 million inhabitants in 2021) on the Saigon River and its estuary and provide a comprehensive water quality assessment. Water samples were collected along a 140-km stretch integrating the river-estuary continuum from upstream HCMC down to the estuary mouth in the East Sea. Additional water samples were collected at the mouth of the four main canals of the city center. Chemical analysis was performed targeting up to 217 micropollutants (pharmaceuticals, plasticizers, PFASs, flame retardants, hormones, pesticides). Bioanalysis was performed using six in-vitro bioassays for hormone receptor-mediated effects, xenobiotic metabolism pathways and oxidative stress response, respectively, all accompanied by cytotoxicity measurement. A total of 120 micropollutants were detected and displayed high variability along the river continuum with total concentration ranging from 0.25 to 78 µg L-1. Among them, 59 micropollutants were ubiquitous (detection frequency ≥ 80 %). An attenuation was observed in concentration and effect profiles towards the estuary. The urban canals were identified as major sources of micropollutants and bioactivity to the river, and one canal (Ben Nghé) exceeded the effect-based trigger values derived for estrogenicity and xenobiotic metabolism. Iceberg modelling apportioned the contribution of the quantified and the unknown chemicals to the measured effects. Diuron, metolachlor, chlorpyrifos, daidzein, genistein, climbazole, mebendazole and telmisartan were identified as main risk drivers of the oxidative stress response and xenobiotic metabolism pathway activation. Our study reinforced the need for improved wastewater management and deeper evaluations of the occurrence and fate of micropollutants in urbanized tropical estuarine environments.

Three years of wastewater surveillance for new psychoactive substances from 16 countries.

The proliferation of new psychoactive substances (NPS) over recent years has made their surveillance complex. The analysis of raw municipal influent wastewater can allow a broader insight into community consumption patterns of NPS. This study examines data from an international wastewater surveillance program that collected and analysed influent wastewater samples from up to 47 sites in 16 countries between 2019 and 2022. Influent wastewater samples were collected over the New Year period and analysed using validated liquid chromatography - mass spectrometry methods. Over the three years, a total of 18 NPS were found in at least one site. Synthetic cathinones were the most found class followed by phenethylamines and designer benzodiazepines. Furthermore, two ketamine analogues, one plant based NPS (mitragynine) and methiopropamine were also quantified across the three years. This work demonstrates that NPS are used across different continents and countries with the use of some more evident in particular regions. For example, mitragynine has highest mass loads in sites in the United States, while eutylone and 3-methylmethcathinone increased considerably in New Zealand and in several European countries, respectively. Moreover, 2F-deschloroketamine, an analogue of ketamine, has emerged more recently and could be quantified in several sites, including one in China, where it is considered as one of the drugs of most concern. Finally, some NPS were detected in specific regions during the initial sampling campaigns and spread to additional sites by the third campaign. Hence, wastewater surveillance can provide an insight into temporal and spatial trends of NPS use.

A novel method for extraction, clean-up and analysis of per- and polyfluoroalkyl substances (PFAS) in different plant matrices using LC-MS/MS.

Per- and polyfluoroalkyl substances (PFAS) are chemicals of concern due to their persistence, bioaccumulation, and toxic properties. PFAS accumulation in plants poses a risk of human and animal exposure due to consumption of the affected plants, but also allows plants to be used in remediation of PFAS-contaminated soils and groundwater. Therefore, effective extraction, cleanup, and analytical methods for measuring PFAS concentrations in plants are fundamental for research on animal and environmental health. PFAS analysis in plant matrices is complex, due to high matrix interference, and scarcity of methods for analyzing different classes of PFAS. In this study, a simple sample preparation method for PFAS analysis in various plant tissues (leaves, needles, twigs, stems, roots from 10 different species) was developed and validated. Instrumental analysis was performed using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The method was optimized considering six different extraction conditions and three different cleanup techniques. Methanol as extraction solvent, combined with 1 g ENVI carb cartridges, showed best performance among all extraction conditions and cleanup techniques tested. Method validation showed good recovery (90-120%), high within-day and between-day precision (<20% relative standard deviation), and low method detection limit (0.04-4.8 ng g-1 dry weight (dw)) for different plant matrices. In tests of the method on soil and different plant tissues of silver birch (Betula pendula) and Norway spruce (Picea abies) at a PFAS-contaminated site, 16 of 24 target PFAS were detected in plants and 17 in soil. Æ©PFAS concentration in soil was 43 ng g-1 dw. PFAS distribution in silver birch tissues ranged from 7.1 ng g-1 dw in roots to 64 ng g-1 dw in leaves, and in Norway spruce from 14 ng g-1 dw in roots to 16 ng g-1 dw in needles. This novel method for PFAS analysis in plants can be valuable in future monitoring, process understanding, remediation, and risk assessments.