Department Environmental Chemistry

The rapid increase in the production and global use of chemicals and their mixtures has raised concerns about their potential impact on human and environmental health. With advances in analytical techniques, in particular, high-resolution mass spectrometry (HRMS), thousands of compounds and transformation products with potential adverse effects can now be detected in environmental samples. However, identifying and prioritizing the toxicity drivers among these compounds remain a significant challenge. Effect-directed analysis (EDA) emerged as an important tool to address this challenge, combining biotesting, sample fractionation, and chemical analysis to unravel toxicity drivers in complex mixtures. Traditional EDA workflows are labor-intensive and time-consuming, hindering large-scale applications. The concept of high-throughput (HT) EDA has recently gained traction as a means of accelerating these workflows. Key features of HT-EDA include the combination of microfractionation and downscaled bioassays, automation of sample preparation and biotesting, and efficient data processing workflows supported by novel computational tools. In addition to microplate-based fractionation, high-performance thin-layer chromatography (HPTLC) offers an interesting alternative to HPLC in HT-EDA. This review provides an updated perspective on the state-of-the-art in HT-EDA, and novel methods/tools that can be incorporated into HT-EDA workflows. It also discusses recent studies on HT-EDA, HT bioassays, and computational prioritization tools, along with considerations regarding HPTLC. By identifying current gaps in HT-EDA and proposing new approaches to overcome them, this review aims to bring HT-EDA a step closer to monitoring applications.

Cationic surfactants are used in many industrial processes and in consumer products with concurrent release into the aquatic environment, where they may accumulate in aquatic organisms to regulatoryly relevant thresholds. Here, we aimed to better understand the bioconcentration behavior of three selected cationic surfactants, namely N,N-dimethyldecylamine (T10), N-methyldodecylamine (S12), and N,N,N-trimethyltetradecylammonium cation (Q14), in the cells of fish liver (RTL-W1) and gill (RTgill-W1) cell lines. We conducted full mass balances for bioconcentration tests with the cell cultures, in which the medium, the cell surface, the cells themselves, and the plastic compartment were sampled and quantified for each surfactant by HPLC MS/MS. Accumulation in/to cells correlated with the surfactants' alkyl chain lengths and their membrane lipid–water partitioning coefficient, D_MLW. Cell-derived bioconcentration factors (BCF) of T10 and S12 were within a factor of 3.5 to in vivo BCF obtained from the literature, while the cell-derived BCF values for Q14 were >100 times higher than the in vivo BCF. From our experiments, rainbow trout cell lines appear as a suitable conservative in vitro screening method for bioconcentration assessment of cationic surfactants and are promising for further testing.

Understanding the impact of human activities on the metabolic state of soil and aquatic environments is of paramount importance to implement measures for maintaining ecosystem services. Variations of natural abundance ¹⁸O/¹⁶O ratios in phosphate have been proposed as proxies for the holistic assessment of metabolic activity given the crucial importance of phosphoryl transfer reactions in fundamental biological processes. However, instrumental and procedural limitations inherent to oxygen isotope analysis in phosphate and organophosphorus compounds have so far limited the stable isotope-based evaluation of metabolic processes. Here, we discuss how recent developments in Orbitrap high resolution mass spectrometry enable measurements of ¹⁸O/¹⁶O ratios in phosphate and outline the critical mass spectrometry parameters for accurate and precise analysis. Subsequently, we evaluate the types of ¹⁸O kinetic isotope effects of phosphoryl transfer reactions and illustrate how novel analytical approaches will give rise to an improved understanding of ¹⁸O/¹⁶O ratio variations from biochemical processes affecting the microbial phosphorus metabolism.

Rieske oxygenases are known as catalysts that enable the cleavage of aromatic and aliphatic C–H bonds in structurally diverse biomolecules and recalcitrant organic environmental pollutants through substrate oxygenations and oxidative heteroatom dealkylations. Yet, the unproductive O₂ activation, which is concomitant with the release of reactive oxygen species (ROS), is typically not taken into account when characterizing Rieske oxygenase function. Even if considered an undesired side reaction, this O₂ uncoupling allows for studying active site perturbations, enzyme mechanisms, and how enzymes evolve as environmental microorganisms adapt their substrates to alternative carbon and energy sources. Here, we report on complementary methods for quantifying O₂ uncoupling based on mass balance or kinetic approaches that relate successful oxygenations to total O₂ activation and ROS formation. These approaches are exemplified with data for two nitroarene dioxygenases (nitrobenzene and 2-nitrotoluene dioxygenase) which have been shown to mono- and dioxygenate substituted nitroaromatic compounds to substituted nitrobenzylalcohols and catechols, respectively.

Oxygenation of aromatic and aliphatic hydrocarbons by Rieske oxygenases is the initial step of various biodegradation pathways for environmental organic contaminants. Microorganisms carrying Rieske oxygenases are able to quickly adapt their substrate spectra to alternative carbon and energy sources that are structurally related to the original target substrate, yet the molecular events responsible for this rapid adaptation are not well understood. Here, we evaluated the hypothesis that reactive oxygen species (ROS) generated by unproductive activation of O₂, the so-called O₂ uncoupling, in the presence of the alternative substrate exert a selective pressure on the bacterium for increasing the oxygenation efficiency of Rieske oxygenases. To that end, we studied wild-type 2-nitrotoluene dioxygenase from Acidovorax sp. strain JS42 and five enzyme variants that have evolved from adaptive laboratory evolution experiments with 3- and 4-nitrotoluene as alternative growth substrates. The enzyme variants showed a substantially increased oxygenation efficiency toward the new target substrates concomitant with a reduction of ROS production, while mechanisms and kinetics of enzymatic O₂ activation remained unchanged. Structural analyses and docking studies suggest that amino acid substitutions in enzyme variants occurred at residues lining both substrate and O₂ transport tunnels, enabling tighter binding of the target substrates in the active site. Increased oxygenation efficiencies measured in vitro for the various enzyme (variant)-substrate combinations correlated linearly with in vivo changes in growth rates for evolved Acidovorax strains expressing the variants. Our data suggest that the selective pressure from oxidative stress toward more efficient oxygenation by Rieske oxygenases was most notable when O₂ uncoupling exceeded 60%.

Abwassereinträge aus der chemisch-pharmazeutischen Industrie wirken sich messbar auf die stoffliche Belastung der ARA-Abläufe aus und hinterlassen eine hochvariable, standortspezifische Signatur. Betriebsspezifische Messkampagnen helfen den Unternehmen, ihre Einträge adäquat zu erfassen und diese mit gezielten Massnahmen abzusenken.

Wirkstoffeinträge von formulierenden Pharmabetrieben führen häufig zu Spitzenkonzentrationen in kommunalen Abwasserreinigungsanlagen. Schweizweite Untersuchungen in Zusammenarbeit mit neun Betrieben zeigen das Ausmass der Einträge für unterschiedliche Abwasservorbehandlungen. Die Studie schliesst wichtige Kenntnislücken bezüglich der Abwasserbelastung von formulierenden Pharmabetrieben und zeigt Handlungsoptionen für die Unternehmen auf.

Herein, we present a new computational methodology that unlocks the prediction of the complex multi-species multi-equilibria processes involved in the formation of complex metal-oxo nanoclusters. Relying on our recently introduced method named POMSimulator, we extended its capabilities and challenged its accuracy with the well-known phosphomolybdate [PMo₁₂O₄₀]³⁻ Keggin anion system. We show how the use of statistical techniques enabled the processing of a vast number of speciation models and their associated systems of non-linear equations efficiently and in a scalable manner. Subsequently, this approach is applied to generate statistically averaged speciation diagrams and their associated error bars. Then, we unveil the previously unreported speciation phase diagram under varying [Mo]/[P] ratios vs. pH. Our findings align well with experimental data, indicating the prevalence of the Keggin {PMo₁₂} as the primary species at low pH, but the lacunary {PMo₁₁}and Strandberg {P₂Mo₅} anions also emerge as major species at other concentration ratios. Finally, from 7 × 10⁴ speciation models we inferred a plausible reaction network across the diverse nuclearities present within the system, which underlines the role of trimers as key intermediate building blocks.

Per- and polyfluoroalkyl substances (PFAS), particularly the perfluorinated ones, are recalcitrant to biodegradation. By integrating an enrichment culture of reductive defluorination with biocompatible electrodes for the electrochemical process, a deeper defluorination of a C₆-perfluorinated unsaturated PFAS was achieved compared to the biological or electrochemical system alone. Two synergies in the bioelectrochemical system were identified: i) The in-series microbial-electrochemical defluorination and ii) the electrochemically enabled microbial defluorination of intermediates. These synergies at the material–microbe interfaces surpassed the limitation of microbial defluorination and further turned the biotransformation end products into less fluorinated products, which could be less toxic and more biodegradable in the environment. This material–microbe hybrid system brings opportunities in the bioremediation of PFAS driven by renewable electricity and warrants future research on mechanistic understanding of defluorinating and electroactive microorganisms at the material–microbe interface for system optimizations.

Catchment urbanisation results in urban streams being exposed to a multitude of stressors. Notably, stressors originating from diffuse sources have received less attention than stressors originating from point sources. Here, advances related to diffuse urban stressors and their consequences for stream benthic communities are summarised by reviewing 92 articles. Based on the search criteria, the number of articles dealing with diffuse urban stressors in streams has been increasing, and most of them focused on North America, Europe, and China. Land use was the most common measure used to characterize diffuse stressor sources in urban streams (70.7 % of the articles characterised land use), and chemical stressors (inorganic nutrients, xenobiotics, metals, and water properties, including pH and conductivity) were more frequently reported than physical or biological stressors. A total of 53.3 % of the articles addressed the impact of urban stressors on macroinvertebrates, while 35.9 % focused on bacteria, 9.8 % on fungi, and 8.7 % on algae. Regarding ecosystem functions, almost half of the articles (43.5 %) addressed changes in community dynamics, 40.3 % addressed organic matter decomposition, and 33.9 % addressed nutrient cycling. When comparing urban and non-urban streams, the reviewed studies suggest that urbanisation negatively impacts the diversity of benthic organisms, leading to shifts in community composition. These changes imply functional degradation of streams. The results of the present review summarise the knowledge gained to date and identify its main gaps to help improve our understanding of urban streams.

La urbanización de las cuencas expone a los arroyos urbanos a multitud de factores de estrés. Destacan aquellos que tienen su origen en fuentes difusas, los cuales han recibido menos atención que aquellos estresores procedentes de fuentes puntuales. Este estudio resume los avances relacionados con los estresores urbanos difusos y sus consecuencias para las comunidades bentónicas fluviales, a partir de la revisión de 92 artículos. En base a los criterios de búsqueda, el número de artículos que tratan sobre estresores urbanos difusos en arroyos ha ido en aumento, la mayoría de ellos centrados en América del Norte, Europa y China. Los usos del suelo fueron la variable más utilizada para caracterizar las fuentes difusas de estrés (el 70.7 % de los artículos caracterizó los usos del suelo), y los factores de estrés químico (nutrientes inorgánicos o propiedades del agua, como pH o conductividad) se mencionaron con más frecuencia que los factores de estrés físico o biológico. El 53.3 % de los trabajos abordaron el impacto de los estresores urbanos difusos sobre los macroinvertebrados, mientras que el 35.9 % se centraron en bacterias, el 9.8 % en hongos y el 8.7 % en algas. En cuanto al funcionamiento de los ecosistemas, prácticamente la mitad de los trabajos (43.5 %) analizó cambios en la dinámica de las comunidades, el 40.3 % en la descomposición de materia orgánica y el 33.9 % en los ciclos de los nutrientes. Al comparar los arroyos urbanos y no urbanos, los trabajos revisados sugieren que la urbanización afecta negativamente a la diversidad de organismos bentónicos, provocando cambios en la composición de la comunidad. Estos cambios implican la degradación funcional de los arroyos. Los resultados de la presente revisión resumen los conocimientos adquiridos hasta hoy e identifican sus principales carencias a fin de ayudar a mejorar nuestra comprensión de los arroyos urbanos.

Environmental risk assessment of chemical contaminants requires prioritizing of substances taken up by biota as it is a starting point for potential adverse effects. Although knowledge about the occurrence of known chemical pollutants in aquatic organisms has significantly improved during the last decade, there is still a poor understanding for a broad range of more polar compounds. To tackle this issue, we proposed an approach that identifies bioaccumulative and biomagnifiable polar chemicals using liquid chromatography coupled with electrospray ionization to high resolution tandem mass spectrometry (LC-HRMS/MS) and combine it with trend analysis using hierarchical clustering. As a proof-of-concept, this approach was implemented on various organisms and compartments (sediment, litter leaves, periphytic biofilm, invertebrates and fish) collected from a small urban river. HRMS/MS data measured via data-independent acquisition mode were retrospectively analysed using two analytical strategies: (1) retrospective target and (2) suspect/non-target screening. In the retrospective target analysis, 56 of 361 substances spanning a broad range of contaminant classes were detected (i.e. 26 in fish, 18 in macroinvertebrates, 28 in leaves, 29 in periphyton and 32 in sediments, with only 7 common to all compartments), among which 49 could be quantified using reference standards. The suspect screening approach based on two suspect lists (in-house, Norman SusDat) led to the confirmation of 5 compounds with standards (three xenobiotics at level 1 and two lipids at level 2) and tentative identification of seven industrial or natural chemicals at level 2 and 3 through a mass spectra library match. Overall, this proof-of-concept study provided a more comprehensive picture of the exposure of biota to emerging contaminants (i.e., the internal chemical exposome) and potential bioaccumulation or biomagnification of polar compounds along the trophic chain.

Der Bundesrat verabschiedete 2017 den Aktionsplan Pflanzenschutzmittel. Die VSA-Plattform «Wasserqualität» hat im Auftrag des Bundesamts für Umwelt (BAFU) evaluiert, ob die Ziele des Aktionsplans für die Fliessgewässer erreicht werden. Es zeigen sich erste Verbesserungen in der Wasserqualität, aber Pflanzenschutzmittel belasten weiterhin Bäche und Flüsse.

Background: prioritisation of chemical pollutants is a major challenge for environmental managers and decision-makers alike, which is essential to help focus the limited resources available for monitoring and mitigation actions on the most relevant chemicals. This study extends the original NORMAN prioritisation scheme beyond target chemicals, presenting the integration of semi-quantitative data from retrospective suspect screening and expansion of existing exposure and risk indicators. The scheme utilises data retrieved automatically from the NORMAN Database System (NDS), including candidate substances for prioritisation, target and suspect screening data, ecotoxicological effect data, physico-chemical data and other properties. Two complementary workflows using target and suspect screening monitoring data are applied to first group the substances into six action categories and then rank the substances using exposure, hazard and risk indicators. The results from the 'target' and 'suspect screening' workflows can then be combined as multiple lines of evidence to support decision-making on regulatory and research actions.
Results: as a proof-of-concept, the new scheme was applied to a combined dataset of target and suspect screening data. To this end, > 65,000 substances on the NDS, of which 2579 substances supported by target wastewater monitoring data, were retrospectively screened in 84 effluent wastewater samples, totalling > 11 million data points. The final prioritisation results identified 677 substances as high priority for further actions, 7455 as medium priority and 326 with potentially lower priority for actions. Among the remaining substances, ca. 37,000 substances should be considered of medium priority with uncertainty, while it was not possible to conclude for 19,000 substances due to insufficient information from target monitoring and uncertainty in the identification from suspect screening. A high degree of agreement was observed between the categories assigned via target analysis and suspect screening-based prioritisation. Suspect screening was a valuable complementary approach to target analysis, helping to prioritise thousands of substances that are insufficiently investigated in current monitoring programmes.
Conclusions: this updated prioritisation workflow responds to the increasing use of suspect screening techniques. It can be adapted to different environmental compartments and can support regulatory obligations, including the identification of specific pollutants in river basins and the marine environments, as well as the confirmation of environmental occurrence levels predicted by modelling tools.

Climate change affects more than elsewhere the northern circumpolar permafrost region. This zone comprises large rivers flowing mainly to the Arctic Ocean, delivering about 10 % of the global riverine water flux. These pan-Arctic Rivers drive the dynamics of northern organic carbon (OC) and mercury (Hg) cycling. Permafrost degradation may release substantial amounts of OC and Hg, with potential regional and global impacts. In this review, we summarise the main findings in the last three decades about the role of the pan-Arctic Rivers in OC and Hg cycling and the effect of climate change on these dynamics. Total DOC and POC fluxes delivered by the pan-Arctic rivers presently reach 34.4 ± 1.2 TgC·yr⁻¹ and 7.9 ± 0.5 TgC·yr⁻¹, while the export of Hg reaches 38.9 ± 1.7 Mg·yr⁻¹. This review highlights future challenges for the scientific community in evaluating spatial and temporal dynamics of the processes involved in OC and Hg cycling in permafrost-affected areas. Permafrost thawing could lead to greater fluxes of OC and Hg with ill-known resulting risks for food chains. Within this context, efforts should be made to study OC effects on Hg methylation. Moreover, assessing the spatial variability of OC and Hg mobilisation and transport within the pan-Arctic watersheds may help understand the future OC and Hg cycling dynamics in the northern circumpolar permafrost region.

Aquatic ecosystems continue to be threatened by chemical pollution. To what extent organisms are able to cope with chemical exposure depends on their ability to display mechanisms of defense across different organs. Among these mechanisms, biotransformation processes represent key physiological responses that facilitate detoxification and reduce the bioaccumulation potential of chemicals. Biotransformation does not only depend on the ability of different organs to display biotransformation enzymes but also on the affinity of chemicals towards these enzymes. In the present study, we explored the ability of different organs and of two freshwater fish to support biotransformation processes through the determination of in vitro phase I and II biotransformation enzyme activity, and their role in supporting intrinsic clearance and the formation of biotransformation products. Three environmentally relevant pollutants were evaluated: the polycyclic aromatic hydrocarbon (PAH) pyrene (as recommended by the OECD 319b test guideline), the fungicide azoxystrobin, and the pharmaceutical propranolol. Comparative studies using S9 sub-cellular fractions derived from the liver, intestine, gills, and brain of brown trout (Salmo trutta) and rainbow trout (Oncorhynchus mykiss) revealed significant phase I and II enzyme activity in all organs. However, organ- and species-specific differences were found. In brown trout, significant extrahepatic biotransformation was observed for pyrene but not for azoxystrobin and propranolol. In rainbow trout, the brain appeared to biotransform azoxystrobin. In this same species, propranolol appeared to be biotransformed by the intestine and gills. Biotransformation products could be detected only from hepatic biotransformation, and their profiles and formation rates displayed species-specific patterns and occurred at different magnitudes. Altogether, our findings further contribute to the current understanding of organ-specific biotransformation capacity, beyond the expression and activity of enzymes, and its dependence on specific enzyme-chemical interactions to support mechanisms of defense against exposure.

Abstract: enviPath is a widely used database and prediction system for microbial biotransformation pathways of primarily xenobiotic compounds. Data and prediction system are freely available both via a web interface and a public REST API. Since its initial release in 2016, we extended the data available in enviPath and improved the performance of the prediction system and usability of the overall system. We now provide three diverse data sets, covering microbial biotransformation in different environments and under different experimental conditions. This also enabled developing a pathway prediction model that is applicable to a more diverse set of chemicals. In the prediction engine, we implemented a new evaluation tailored towards pathway prediction, which returns a more honest and holistic view on the performance. We also implemented a novel applicability domain algorithm, which allows the user to estimate how well the model will perform on their data. Finally, we improved the implementation to speed up the overall system and provide new functionality via a plugin system.
Scientific contribution: The main scientific contributions are the development of a pathway prediction model applicable to diverse chemicals, a specialized evaluation method for holistic performance assessment, and a novel applicability domain algorithm for user-specific performance estimation. The introduction of two new data sets, and the creation of links to EC classes make enviPath a unique resource in microbial biotransformation research.

Separate collection and treatment of urine optimizes nutrient recovery and enhances micropollutant removal from municipal wastewater. One typical urine treatment train includes nutrient recovery in three biological processes: anaerobic storage, followed by aerobic organics degradation concurrently with nitrification. These are usually followed by activated carbon adsorption to remove micropollutants. However, removing micropollutants prior to nitrification would protect nitrifiers from potential inhibition by pharmaceuticals. In addition, combining simplified biological treatment with activated carbon adsorption could offer a cheap and robust process for removing micropollutants where nutrient recovery is not the first priority, as a partial loss of ammonia occurs without nitrification. In this study, we investigated whether activated carbon adsorption could also take place between the three biological treatment steps. We tested the effectiveness of micropollutant removal with activated carbon after each biological treatment step by conducting experiments with anaerobically stored urine, organics-depleted urine, and nitrified urine. The urine solutions were spiked with 19 pharmaceuticals: amisulpride, atenolol, atenolol acid, candesartan, carbamazepine, citalopram, clarithromycin, darunavir, diclofenac, emtricitabine, fexofenadine, hydrochlorothiazide, irbesartan, lidocaine, metoprolol, N₄-acetylsulfamethoxazole, sulfamethoxazole, trimethoprim, venlafaxine, and two artificial sweeteners, acesulfame and sucralose. Batch experiments were conducted with powdered activated carbon (PAC) to determine how much activated carbon achieve which degree of micropollutant removal and how organics, pH, and speciation change from ammonium to nitrate influence adsorption. Micropollutant removal was also tested in granular activated carbon (GAC) columns, which is the preferred technology for micropollutant removal from urine. The carbon usage rates (CUR) per person were lower for all urine solutions than for municipal wastewater. The results showed that organics depletion would be needed when micropollutant removal was the sole aim of urine treatment, as the degradation of easily biodegradable organics prevented clogging of GAC columns. However, CUR did hardly improve with organics-depleted urine compared to stored urine. The lowest CUR was achieved with nitrified urine. This resulted from the additional organics removal during nitrification and not the lower pH or the partial conversion of ammonium to nitrate. In addition, we showed that the relative pharmaceutical removal in all solutions was independent of the initial pharmaceutical concentration unless the background organics matrix changed considerably. We conclude that removal of micropollutants in GAC columns from organics-depleted urine can be performed without clogging, but with the drawback of a higher carbon usage compared to removal from nitrified urine.

The management and mitigation of chemical pollution are key elements of sustainable development initiatives that aim to provide safe and clean water. While environmental scientists are developing the capabilities to assess the fate, (eco)toxicity and risks of a plethora of synthetic chemicals comprehensively, notorious pollution scenarios and decontamination challenges call for targeted and case-specific evaluation of chemical hazards. Here we review the utility and perspectives of compound-specific isotope analysis for obtaining an understanding of environmental processes that allows one to identify pollution sources, assess contaminant (bio)transformation and gain insights into reaction pathways. Using three prototypical scenarios of water contamination, namely point-source pollution of groundwater at contaminated sites, diffuse pollution of soils and surface waters through pesticide use and the abatement of pharmaceuticals and disinfection by-products in water treatment systems, we illustrate both success stories of compound-specific isotope analysis and current developments to address challenges for future applications.

Die zunehmende Produktion und Verwendung von Chemikalien hat zu großem Interesse am Monitoring neuer Stoffe durch Suspect- und Non-Target-Screening (NTS) geführt. Um die Umsetzung von NTS in wissenschaftlichen, kommerziellen und staatlichen Laboren sowie die Akzeptanz durch Behörden zu erleichtern, ist eine Harmonisierung von NTS erforderlich. Hierfür haben Mitglieder der Norman Association (Network of reference laboratories, research centres, and related organizations for monitoring of emerging environmental substances) einen Leitfaden erstellt. Das Dokument soll als Richtlinie für die Durchführung qualitativ hochwertiger NTS-Studien dienen und gleichzeitig das Bewusstsein für die Möglichkeiten, aber auch die Grenzen schärfen. Es enthält Anleitungen für alle Schritte, von der Probenahme und -aufbereitung über die Chromatographie (Flüssig- und Gaschromatographie – (LC und GC), verschiedene Ionisierungstechniken gekoppelt an die hochauflösende Tandem-Massenspektrometrie (HRMS/MS), bis hin zur Datenauswertung und Berichterstattung.

Through the application of C-CF₃-containing plant protection products (PPP) in agriculture, a substantial quantity of trifluoroacetate (TFA) can be formed and emitted. We here present estimations of TFA formation potentials from PPP across three important economical regions, namely Europe, the United States of America and China. PPP with TFA formation potential vary in type and use profile across those regions, but can be found throughout, with the estimated maximum TFA emissions ranging from 0 to 83 kg/km² per year. Therein, some PPP are only used for specific crops in specific regions, while others are used more widely. The importance of PPP as a TFA source is supported by the field data from a region in Germany, which revealed a significant increase in TFA groundwater concentrations with agriculture compared to other land uses. Substance-specific TFA formation rates and field studies are necessary to characterize the formation of TFA from precursors under environmental conditions and to rank and prioritize PPP of concern for potential (regulatory) action.

Wastewater Treatment Plants (WWTP) are a major repository and entrance path of nanoparticles (NP) in the environment and hence play a major role in the final NP fate and toxicity. Studies on silver nanoparticles (AgNP) transport via the WWTP system and uptake by aquatic organisms have so far been carried out using unrealistically high AgNP concentrations, unlikely to be encountered in the aquatic environment. The use of high AgNP concentrations is necessitated by both the low sensitivity of the detection methods used and the need to distinguish background Ag from spiked AgNP.
In this study, isotopically enriched ¹⁰⁹AgNP were synthesized to overcome these shortcomings and characterized by a broad range of methods including transmission electron microscopy, dynamic and electrophoretic light scattering. ¹⁰⁹AgNP and gold NP (AuNP) were spiked to a pilot wastewater treatment plant fed with municipal wastewater for up to 21 days. AuNP were used as chemically less reactive tracer. The uptake of the pristine and transformed NP present in the effluent was assessed using the benthic amphipod Hyalella azteca in fresh- and brackish water exposures at environmentally relevant concentrations of 30 to 500 ng Au/L and 39 to 260 ng Ag/L. The unique isotopic signature of the ¹⁰⁹AgNP allowed to detect the material at environmentally relevant concentrations in the presence of a much higher natural Ag background. The results show that the transformations reduce the NP uptake at environmentally relevant exposure concentrations. For ¹⁰⁹Ag, lower accumulation factors (AF) were obtained after exposure to transformed NP (250–350) compared to the AF values obtained for pristine ¹⁰⁹AgNP (750–840). The reduced AF values observed for H. azteca exposed to effluent from the AuNP-spiked WWTP indicate that biological transformation processes (e.g. eco-corona formation) seem to be involved in addition to chemical transformation.

Non-targeted and suspect screening analysis using liquid chromatography coupled to high-resolution mass spectrometry (LC-HRMS) holds great promise to comprehensively characterize complex chemical mixtures. Data preprocessing is a crucial part of the process, however, some limitations are observed: (i) peak-picking and feature extraction might be incomplete, especially for low abundant compounds, and (ii) limited reproducibility has been observed between laboratories and software for detected features and their relative quantification. We first conducted a critical review of existing solutions that could improve the reproducibility of preprocessing for LC-HRMS. Solutions include providing repositories and reporting guidelines, open and modular processing workflows, public benchmark datasets, tools to optimize the data preprocessing and to filter out false positive detections. We then propose harmonized quality assurance/quality control guidelines that would allow to assess the sensitivity of feature detection, reproducibility, integration accuracy, precision, accuracy, and consistency of data preprocessing for human biomonitoring, food and environmental communities.

Nontargeted screening (NTS) utilizing liquid chromatography electrospray ionization high-resolution mass spectrometry (LC/ESI/HRMS) is increasingly used to identify environmental contaminants. Major differences in the ionization efficiency of compounds in ESI/HRMS result in widely varying responses and complicate quantitative analysis. Despite an increasing number of methods for quantification without authentic standards in NTS, the approaches are evaluated on limited and diverse data sets with varying chemical coverage collected on different instruments, complicating an unbiased comparison. In this interlaboratory comparison, organized by the NORMAN Network, we evaluated the accuracy and performance variability of five quantification approaches across 41 NTS methods from 37 laboratories. Three approaches are based on surrogate standard quantification (parent-transformation product, structurally similar or close eluting) and two on predicted ionization efficiencies (RandFor-IE and MLR-IE). Shortly, HPLC grade water, tap water, and surface water spiked with 45 compounds at 2 concentration levels were analyzed together with 41 calibrants at 6 known concentrations by the laboratories using in-house NTS workflows. The accuracy of the approaches was evaluated by comparing the estimated and spiked concentrations across quantification approaches, instrumentation, and laboratories. The RandFor-IE approach performed best with a reported mean prediction error of 15× and over 83% of compounds quantified within 10× error. Despite different instrumentation and workflows, the performance was stable across laboratories and did not depend on the complexity of water matrices.

Pharmaceuticals and their human metabolites are contaminants of emerging concern in the aquatic environment. Most monitoring studies focus on a limited set of parent compounds and even fewer metabolites. However, more than 50% of the most consumed pharmaceuticals are excreted in higher amounts as metabolites than as parents, as confirmed by a literature analysis within this study. Hence, we applied a wide-scope suspect screening approach to identify human pharmaceutical metabolites in wastewater influent from three Swiss treatment plants. Based on consumption amounts and human metabolism data, a suspect list comprising 268 parent compounds and over 1500 metabolites was compiled. Online solid phase extraction combined with liquid chromatography coupled to high-resolution tandem mass spectrometry was used to analyze the samples. Data processing, annotation, and structure elucidation were achieved with various tools, including molecular networking as well as SIRIUS/CSI:FingerID and MetFrag for MS2 spectra rationalization. We confirmed 37 metabolites with reference standards and 16 by human liver S9 incubation experiments. More than 25 metabolites were detected for the first time in influent wastewater. Semiquantification with MS2Quant showed that metabolite to parent concentration ratios were generally lower compared to literature expectations, probably due to further metabolite transformation in the sewer system or limitations in the metabolite detection. Nonetheless, metabolites pose a large fraction to the total pharmaceutical contribution in wastewater, highlighting the need for metabolite inclusion in chemical risk assessment.

Micropollutants removal efficiency strongly vary across different aerobic wastewater treatment plants, resulting in their frequent detection in surface and ground waters. Seasonal temperature variation is a major factor influencing plant performance, but it is still unclear how prolonged periods of temperature change impact microbiome and micropollutant biotransformation. This work investigates the effect of long-term temperature variation on the microbial dynamics in an activated sludge system, and the impact on micropollutant biotransformation. Sequencing batch reactors were used as model system and 4–40 °C temperature range was studied. 16S rRNA amplicon sequencing showed that temperature drives microbial structure (gDNA) and activity (RNA), rather than time, and this was stronger below 15 °C and above 25 °C. The microbial community was richest and more diverse at 20 °C, while rarer and more specific taxa became predominant over time, at more extreme temperatures. This suggested that less abundant taxa might be responsible for maintaining the biotransformation capability in the activated sludge at extreme temperatures. Micropollutant biotransformation rates mostly deviated from the classic Arrhenius model below 15 °C and above 25 °C, indicating that prolonged exposure to temperature changes leads to temperature-induced taxonomic shifts, resulting in the emerging of different sets of biotransformation pathways over different temperature ranges.

Phytoplankton is an essential resource in aquatic ecosystems, situated at the base of aquatic food webs. Plastic pollution can impact these organisms, potentially affecting the functioning of aquatic ecosystems. The interaction between plastics and phytoplankton is multifaceted: while microplastics can exert toxic effects on phytoplankton, plastics can also act as a substrate for colonisation. By reviewing the existing literature, this study aims to address pivotal questions concerning the intricate interplay among plastics and phytoplankton/phytobenthos and analyse impacts on fundamental ecosystem processes (e.g. primary production, nutrient cycling). This investigation spans both marine and freshwater ecosystems, examining diverse organisational levels from subcellular processes to entire ecosystems. The diverse chemical composition of plastics, along with their variable properties and role in forming the "plastisphere", underscores the complexity of their influences on aquatic environments. Morphological changes, alterations in metabolic processes, defence and stress responses, including homoaggregation and extracellular polysaccharide biosynthesis, represent adaptive strategies employed by phytoplankton to cope with plastic-induced stress. Plastics also serve as potential habitats for harmful algae and invasive species, thereby influencing biodiversity and environmental conditions. Processes affected by phytoplankton–plastic interaction can have cascading effects throughout the aquatic food web via altered bottom-up and top-down processes. This review emphasises that our understanding of how these multiple interactions compare in impact on natural processes is far from complete, and uncertainty persists regarding whether they drive significant alterations in ecological variables. A lack of comprehensive investigation poses a risk of overlooking fundamental aspects in addressing the environmental challenges associated with widespread plastic pollution.

Elucidating the speciation (in terms of concentration versus pH) and understanding the formation mechanisms of polyoxometalates remains a significant challenge, both in experimental and computational domains. POMSimulator is a new methodology that tackles this problem from a purely computational perspective. The methodology uses results from quantum mechanics based methods to automatically set up the chemical reaction network, and to build speciation models. As a result, it becomes possible to predict speciation and phase diagrams, as well as to derive new insights into the formation mechanisms of large molecular clusters. In this work we present the main features of the first open-source version of the software. Since the first report [Chem. Sci. 2020, 11, 8448-8456], POMSimulator has undergone several improvements to keep up with the growing challenges that were tackled. After four years of research, we recognize that the source code is sufficiently stable to share a polished and user-friendly version. The Python code, manual, examples, and install instructions can be found at https://github.com/petrusen/pomsimulator.

Urban expansion leads to increasing water pollution, impacting both human health and ecosystems. This decline in water quality often stems from insufficient wastewater treatment, along with runoff from both urban and agricultural areas. Water quality degradation challenges our efforts for sustainable water management and hinders progress toward the UN's Sustainable Development Goals (SDGs), particularly SDG6. Within aquatic environments, the hyporheic zone—the subsurface area where surface water and groundwater mix—plays a crucial role in facilitating pollutant turnover and overall aquatic health. Mixing between surface water and groundwater generates diverse microhabitats in the streambed with varying levels of oxygen, temperature, and chemical composition, which in turn allows diverse microbial communities to strive. The complexity of exchange flows within the hyporheic zone and associated turnover processes of pollutants and nutrients can only be assessed with detailed, cross-disciplinary data sets including data about hydrology, climatology, biogeochemistry, and the subsurface composition. However, integrated data sets of this kind are seldom available. As a result, the drivers behind pollutant dynamics in stream-aquifer systems are still not fully understood. Addressing this knowledge deficit, we present a comprehensive unique data set from an urban stream-aquifer system in Switzerland spanning over 6 months. Incorporating hydrometric, tracer, nutrient, microbial and organic micropollutant data, our data set can help to shed light on the intricate mechanisms governing hyporheic exchange flows, as well as nutrient and organic micropollutant cycling in urban environments.

Cyanobacterial blooms occur at increasing frequency and intensity, notably in freshwater. This leads to the introduction of complex mixtures of their products, i.e., cyano-metabolites, to drinking water treatment plants. To assess the fate of cyano-metabolite mixtures during ozonation, a novel multicompound ozone (O₃) competition kinetics method was developed. Sixteen competitors with known second-order rate constants for their reaction with O₃ ranging between 1 and 10⁸ M^–1 s^–1 were applied to cover a wide range of the O₃ reactivity. The apparent second-order rate constants (k_app,O3) at pH 7 were simultaneously determined for 31 cyano-metabolites. k_app,O3 for olefin- and phenol-containing cyano-metabolites were consistent with their expected reactivity (0.4–1.7 × 10⁶ M^–1 s^–1) while k_app,O3 for tryptophan- and thioether-containing cyano-metabolites were significantly higher than expected (3.4–7.3 × 10⁷ M^–1 s^–1). Cyano-metabolites containing these moieties are predicted to be well abated during ozonation. For cyano-metabolites containing heterocycles, k_app,O3 varied from <10² to 5.0 × 10³ M^–1 s^–1, giving first insights into the O₃ reactivity of this class of compounds. Due to lower O₃ reactivities, heterocycle- and aliphatic amine-containing cyano-metabolites may be only partially degraded by a direct O₃ reaction near circumneutral pH. Hydroxyl radicals, which are formed during ozonation, may be more important for their abatement. This novel multicompound kinetic method allows a high-throughput screening of ozonation kinetics.

Some bacteria can degrade organic micropollutants (OMPs) as primary carbon sources. Due to typically low OMP concentrations, these bacteria may benefit from supplemental assimilation of natural substrates present in the pool of dissolved organic matter (DOM). The biodegradability of such auxiliary substrates and the impacts on OMP removal are tightly linked to biotransformation pathways. Here, we aimed to elucidate the biodegradability and effect of different DOM constituents for the carbofuran degrader Novosphingobium sp. KN65.2, using a novel approach that combines pathway prediction, laboratory experiments, and fluorescence spectroscopy. Pathway prediction suggested that ring hydroxylation reactions catalysed by Rieske-type dioxygenases and flavin-dependent monooxygenases determine the transformability of the 11 aromatic compounds used as model DOM constituents. Our approach further identified two groups with distinct transformation mechanisms amongst the four growth-supporting compounds selected for mixed substrate biodegradation experiments with the pesticide carbofuran (Group 1: 4-hydroxybenzoic acid, 4-hydroxybenzaldehyde; Group 2: p-coumaric acid, ferulic acid). Carbofuran biodegradation kinetics were stable in the presence of both Group 1 and Group 2 auxiliary substrates. However, Group 2 substrates would be preferable for bioremediation processes, as they showed constant biodegradation kinetics under different experimental conditions (pre-growing KN65.2 on carbofuran vs. DOM constituent). Furthermore, Group 2 substrates were utilisable by KN65.2 in the presence of a competitor (Pseudomonas fluorescens sp. P17). Our study thus presents a simple and cost-efficient approach that reveals mechanistic insights into OMP-DOM biodegradation.

Generally, karst aquifers and springs are highly susceptible to contamination due to the high permeability and, therefore, groundwater flow velocities. The often thin soil cover, accompanied by dolines, can lead to fast infiltration of precipitation water loaded with mobilized contaminants such as pesticides and their transformation products. To date, continuous, temporally highly resolved in-situ monitoring to decipher concentration dynamics for a broad range of pesticides is missing. Therefore, a transportable HPLC-HRMS/MS system (MS²field) was positioned at two karst study sites in the Swiss Jura. Water samples were collected and analyzed for pesticides and their transformation products in-situ every 20 min for 6 weeks in 2021 and 8 weeks in 2022. During the spraying season in 2021, six rain events at site 1 and three at site 2 in 2022 were captured. Concurrently, the water quality parameters electrical conductivity, pH, nitrate, turbidity, and water level, were monitored continuously at high temporal resolution. Further, bacterial cell counts were monitored via online flow cytometry.
In 2021, several pesticides and pesticide transformation products were detected in peak concentrations after rain events, of which metamitron showed the highest concentration of up to 1000 ng/L. In one rain event, the Swiss federal and EU drinking water limit of 100 ng/L was exceeded for up to 38 h. Compared with highly frequent MS²field samples collected every 20 min, 42-hours composite samples severely underestimated peak concentrations for all compounds, especially for labile ones. Therefore, it was demonstrated that exceedences of the regulatory limit would have been missed if just composite sampling would have been conducted. Peak concentrations of pesticides coincided with peaks in nitrate concentration and bacterial cell counts following rain events. The correlation analysis showed strong correlations between the three analyzed contaminants (pesticides, nitrate and bacteria), and the proxy parameters electrical conductivity, and pH. The investigation of a second spring revealed similar dynamics indicating that these can be expected in other karst aquifers as well.

Trace organic contaminants (TrOCs) are omnipresent in wastewater treatment plants (WWTPs), yet, their removal during wastewater treatment is oftentimes incomplete and underlying biotransformation mechanisms are not fully understood. In this study, we elucidate how different factors, including pre-exposure levels and duration, influence microbial adaptation towards catabolic TrOC biodegradation and its potential role in biological wastewater treatment. Four sequencing batch reactors (SBRs) were operated in parallel in three succeeding phases, adding and removing a selection of 26 TrOCs at different concentration levels. After each phase of SBR operation, a series of batch experiments was conducted to monitor biotransformation kinetics of those same TrOCs across various spike concentrations. For half of our test TrOCs, we detected increased biotransformation in sludge pre-exposed to TrOC concentrations ≥5 µg L⁻¹ over a 30-day period, with most significant differences observed for the insect repellent DEET and the artificial sweetener saccharin. Accordingly, 16S rRNA amplicon sequencing revealed enrichment of taxa that have previously been linked to catabolic biodegradation of several test TrOCs, e.g., Bosea sp. and Shinella sp. for acesulfame degradation, and Pseudomonas sp. for caffeine, cyclamate, DEET, metformin, paracetamol, and isoproturon degradation. We further conducted shotgun metagenomics to query for gene products previously reported to be involved in the TrOCs’ biodegradation pathways. In the future, directed microbial adaptation may be a solution to improve bioremediation of TrOCs in contaminated environments or in WWTPs.

The UV–vis absorbance spectra, molar extinction coefficients and circular dichroism spectra, as well as NMR and high resolution tandem mass spectrometry spectra were determined for two prominent secondary metabolites from cyanobacteria, namely anabaenopeptin A and anabaenopeptin B. The compounds were extracted from the cyanobacterium Planktothrix rubescens CBT929 and purified by flash chromatography and HPLC. Exact amounts of isolated compounds were assessed by quantitative ¹H-NMR with internal calibrant ethyl 4-(dimethylamino)benzoate in DMSO‑d₆ at 298 K with a recycle delay (d1) of 120 s. UV–vis absorbance spectra were recorded in methanol at room temperature. Molar extinction coefficients were determined at 278 nm as 4190 M⁻¹ cm⁻¹ and 2300 M⁻¹ cm⁻¹ in methanol for anabaenopeptin A and anabaenopeptin B, respectively. Circular dichroism spectra and secondary fragmentation mass spectra are also reported.

Computational methods are playing an increasingly important role as a complement to conventional data evaluation methods in analytical chemistry, and particularly mass spectrometry. Computational mass spectrometry (CompMS) is the application of computational methods on mass spectrometry data. Herein, advances in CompMS for small molecule chemistry are discussed in the areas of spectral libraries, spectrum prediction, and tentative structure identification (annotation): Automatic spectrum curation is facilitating the expansion of openly available spectral libraries, a crucial resource both for compound annotation directly and as a resource for machine learning algorithms. Spectrum prediction and molecular fingerprint prediction have emerged as two key approaches to compound annotation. For both, multiple methods based on classical machine learning and deep learning have been developed. Driven by advances in deep learning-based generative chemistry, de novo structure generation from fragment spectra is emerging as a new field of research. This review highlights key publications in these fields, including our approaches RMassBank (automatic spectrum curation) and MSNovelist (de novo structure generation).

The Joint Danube Survey (JDS) is a regular transnational survey to monitor the quality of the Danube and its main tributaries, in accordance with the EU Water Framework Directive. During the JDS4 in 2019, conventional methods to monitor selected biological quality elements were complemented with DNA metabarcoding.
All together 72 phytobenthos samples were collected along the Danube and its major tributaries within the JDS4, using light microscopy and DNA metabarcoding amplifying a fragment of the rbcL marker gene. The (i) applicability of DNA metabarcoding to identify diatom communities compared to microscopy; (ii) diversity metrics between DNA metabarcoding and microscopy analysis and (iii) the usability of DNA metabarcoding for routine monitoring and assessment of the Danube under future JDS surveys were investigated.
Diatom communities resulting from light microscopy and DNA metabarcoding assessments share 26.5% of all taxa which corresponds to 64.3% when considering relative abundances. Discrepancies originate from biases both from metabarcoding, e.g. missing taxa from the reference library, and from microscopy, e.g. overlooking of hardly visible taxa. Microscopy detected more taxa in total but metabarcoding revealed a higher alpha diversity, detecting also very rare taxa in a given sample. Molecular and microscopy based Specific Pollution Sensitivity Index (IPS) values correlated significantly but differences were detected at several sites due to the differences in community composition and the overestimation of large taxa by metabarcoding. Although both methods showed a decreasing trend of IPS along the Danube, the metabarcoding based IPS covered a higher range of quality classes indicating lower values for downstream sites and the tributaries.
We suggest that metabarcoding provides a standardisable and efficient tool in biomonitoring, being more distinctive among quality classes than microscopy. Due to the high sequencing depth, it is able to detect a higher diversity on genetic level in a time- and cost-efficient manner that should be implemented in future quality assessment tools. We recommend its use in future biomonitoring surveys, for now, as a complementary method to conventional ones.

Biodegradation plays a key role in the fate of chemicals in the environment. The variability of biodegradation in time can cause uncertainty in evaluating the environmental persistence and risk of chemicals. However, the seasonality of biodegradation in rivers has not yet been the subject of environmentally relevant testing and systematic investigation for large numbers of chemicals. In this work, we studied the biodegradation of 96 compounds during four seasons at four locations (up- and downstream of WWTPs located on two Swedish rivers). Significant seasonality (ANOVA, p < 0.05) of the first-order rate constant for primary biodegradation was observed for most compounds. Variations in pH and total bacterial cell count were not the major factors explaining the seasonality of biodegradation. Deviation from the classical Arrhenius-type behavior was observed for most of the studied compounds, which calls into question the application of this relationship to correct biodegradation rate constants for differences in environmental temperature. Similarities in magnitude and seasonality of biodegradation rate constants were observed for some groups of chemicals possessing the same functional groups. Moreover, reduced seasonality of biodegradation was observed downstream of WWTPs, while biodegradation rates of most compounds were not significantly different between up- and downstream.

Biodegradation is one of the most important processes influencing the fate of organic contaminants in the environment. Quantitative understanding of the spatial variability in environmental biodegradation is still largely uncharted territory. Here, we conducted modified OECD 309 tests to determine first-order biodegradation rate constants for 97 compounds in 18 freshwater river segments in five European countries: Sweden, Germany, Switzerland, Spain, and Greece. All but two of the compounds showed significant spatial variability in rate constants across European rivers (ANOVA, P < 0.05). The median standard deviation of the biodegradation rate constant between rivers was a factor of 3. The spatial variability was similar between pristine and contaminated river segments. The longitude, total organic carbon, and clay content of sediment were the three most significant explanatory variables for the spatial variability (redundancy analysis, P < 0.05). Similarities in the spatial pattern of biodegradation rates were observed for some groups of compounds sharing a given functional group. The pronounced spatial variability presents challenges for the use of biodegradation simulation tests to assess chemical persistence. To reflect the variability in the biodegradation rate, the modified OECD 309 test would have to be repeated with water and sediment from multiple sites.

The chemical burden on the environment and human population is increasing. Consequently, regulatory risk assessment must keep pace to manage, reduce, and prevent adverse impacts on human and environmental health associated with hazardous chemicals. Surveillance of chemicals of known, emerging, or potential future concern, entering the environment-food-human continuum is needed to document the reality of risks posed by chemicals on ecosystem and human health from a one health perspective, feed into early warning systems and support public policies for exposure mitigation provisions and safe and sustainable by design strategies. The use of less-conventional sampling strategies and integration of full-scan, high-resolution mass spectrometry and effect-directed analysis in environmental and human monitoring programmes have the potential to enhance the screening and identification of a wider range of chemicals of known, emerging or potential future concern. Here, we outline the key needs and recommendations identified within the European Partnership for Assessment of Risks from Chemicals (PARC) project for leveraging these innovative methodologies to support the development of next-generation chemical risk assessment.

The mass proliferation of cyanobacteria, episodes known as blooms, is a concern worldwide. One of the most critical aspects during these blooms is the production of toxic secondary metabolites that are not limited to the four cyanotoxins recognized by the World Health Organization. These metabolites comprise a wide range of structurally diverse compounds that possess bioactive functions. Potential human and ecosystem health risks posed by these metabolites and co-produced mixtures remain largely unknown. We studied acute lethal and sublethal effects measured as impaired mobility on the freshwater microcrustaceans Thamnocephalus platyurus for metabolite mixtures from two cyanobacterial strains, a microcystin (MC) producer and a non-MC producer. Both cyanobacterial extracts, from the MC-producer and non-MC-producer, caused acute toxicity with LC₅₀ (24 h) values of 0.50 and 2.55 mg_{dw_biomass}/mL, respectively, and decreased locomotor activity. Evaluating the contribution of different cyanopeptides revealed that the Micropeptin-K139-dominated fraction from the MC-producer extract contributed significantly to mortality and locomotor impairment of the microcrustaceans, with potential mixture effect with other cyanopeptolins present in this fraction. In the non-MC-producer extract, compounds present in the apolar fraction contributed mainly to mortality, locomotor impairment, and morphological changes in the antennae of the microcrustacean. No lethal or sublethal effects were observed in the fractions dominated by other cyanopetides (Cyanopeptolin 959, Nostoginin BN741). Our findings contribute to the growing body of research indicating that cyanobacterial metabolites beyond traditional cyanotoxins cause detrimental effects. This underscores the importance of toxicological assessments of such compounds, also at sublethal levels.

Cyanobacterial blooms require monitoring, as they pose a threat to ecosystems and human health, especially by the release of toxins. Along with widely reported microcystins, cyanobacteria coproduce other bioactive metabolites; however, information about their dynamics in surface waters is sparse. We investigated dynamics across full bloom successions throughout a five-year lake monitoring campaign (Greifensee, Switzerland) spanning 150 sampling dates. We conducted extensive suspect screening of cyanobacterial metabolites using the database CyanoMetDB. Across all 850 samples, 35 metabolites regularly co-occurred. Microcystins were present in 70% of samples, with [d-Asp³,(E)-Dhb⁷]MC-RR reaching concentrations of 70 ng/L. Anabaenopeptins, meanwhile, were detected in 95% of all samples with concentrations of Oscillamide Y up to 100-fold higher than microcystins. Based on LC-MS response and frequency, we identified indicator metabolites exclusively produced by one of three cyanobacteria isolated from the lake, these being [d-Asp³,(E)-Dhb⁷]MC-RR from Planktothrix sp. G2020, Microginin 761B from Microcystis sp. G2011, and Ferintoic acid B from Microcystis sp. G2020. These indicators showed distinct temporal trends and peaking seasons that reflect the variance in either the abundance of the producing cyanobacteria or their toxin production dynamics. Our approach demonstrates that selecting high LC-MS response and frequent and species-specific indicator metabolites can be advantageous for cyanobacterial monitoring.

Das Konzept zur Überprüfung des Reinigungseffekts und die 12 Leitsubstanzen haben sich im Vollzug bewährt. Es lässt einen gewissen Spielraum zu bei der Substanzauswahl, was die Behörden nutzen. Für den Reinigungseffekt fällt ins Gewicht, welche Substanzen in die Berechnung einfliessen.

Enzymatic cleavage of C-F bonds in per-and polyfluoroalkyl substances (PFAS) is largely unknown but avidly sought to promote systems biology for PFAS bioremediation. Here, we report the reductive defluorination of α, β-unsaturated per-and polyfluorocarboxylic acids by Acetobacterium spp. The microbial defluorination products were structurally confirmed and showed regiospecificity and stereospecificity, consistent with their formation by enzymatic reactions. A comparison of defluorination activities among several Acetobacterium species indicated that a functional fluoride exporter was required for the detoxification of the released fluoride. Results from both in vivo inhibition tests and in silico enzyme modeling suggested the involvement of enzymes of the flavin-based electron-bifurcating caffeate reduction pathway [caffeoyl-CoA reductase (CarABCDE)] in the reductive defluorination. This is a report on specific microorganisms carrying out enzymatic reductive defluorination of PFAS, which could be linked to electron-bifurcating reductases that are environmentally widespread.

Organic contaminants enter aquatic ecosystems from various sources, including wastewater treatment plant effluent. Freshwater biofilms play a major role in the removal of organic contaminants from receiving water bodies, but knowledge of the molecular mechanisms driving contaminant biotransformations in complex stream biofilm (periphyton) communities remains limited. Previously, we demonstrated that biofilms in experimental flume systems grown at higher ratios of treated wastewater (WW) to stream water displayed an increased biotransformation potential for a number of organic contaminants. We identified a positive correlation between WW percentage and biofilm biotransformation rates for the widely-used insect repellent, N,N-diethyl-meta-toluamide (DEET) and a number of other wastewater-borne contaminants with hydrolyzable moieties. Here, we conducted deep shotgun sequencing of flume biofilms and identified a positive correlation between WW percentage and metagenomic read abundances of DEET hydrolase (DH) homologs. To test the causality of this association, we constructed a targeted metagenomic library of DH homologs from flume biofilms. We screened our complete metagenomic library for activity with four different substrates, including DEET, and a subset thereof with 183 WW-related organic compounds. The majority of active hydrolases in the metagenomic library preferred aliphatic and aromatic ester substrates while, remarkably, only a single reference enzyme was capable of DEET hydrolysis. Of the 626 total enzyme-substrate combinations tested, approximately 5% were active enzyme-substrate pairs. Metagenomic DH family homologs revealed a broad substrate promiscuity spanning 22 different compounds when summed across all enzymes tested. We biochemically characterized the most promiscuous and active enzymes identified based on metagenomic analysis from uncultivated Rhodospirillaceae and Planctomycetaceae. In addition to characterizing new DH family enzymes, we exemplified a framework for linking metagenome-guided hypothesis generation with experimental validation. Overall, this study expands the scope of known enzymatic contaminant biotransformations for metagenomic hydrolases from WW-receiving stream biofilm communities.

When chemical pollutants enter the environment, they can undergo diverse transformation processes, forming a wide range of transformation products (TPs), some of them benign and others more harmful than their precursors. To date, the majority of TPs remain largely unrecognized and unregulated, particularly as TPs are generally not part of routine chemical risk or hazard assessment. Since many TPs formed from oxidative processes are more polar than their precursors, they may be especially relevant in the context of persistent, mobile, and toxic (PMT) and very persistent and very mobile (vPvM) substances, which are two new hazard classes that have recently been established on a European level. We highlight herein that as a result, TPs deserve more attention in research, chemicals regulation, and chemicals management. This perspective summarizes the main challenges preventing a better integration of TPs in these areas: (1) the lack of reliable high-throughput TP identification methods, (2) uncertainties in TP prediction, (3) inadequately considered TP formation during (advanced) water treatment, and (4) insufficient integration and harmonization of TPs in most regulatory frameworks. A way forward to tackle these challenges and integrate TPs into chemical management is proposed.

Background: With the influence of climate change, environmental pollution, global industrialization, and new agricultural practices, increasing amounts of chemical substances with potential risks—both anthropogenic and biogenic—enter the food supply chain, entailing new challenges to food safety and security. Although some food-risk components (FRCs) have been accessed and regulated, the toxicity and exposure level of the numerous components detected in food remain unknown, leaving questions on their effect on food safety. Therefore, multiple databases on emerging FRCs have been constructed to aid in food safety assessment, regulation, and communication; however, their focus areas, data content, quality, and accessibility have not been systematically summarized, which hinders their applications and the development of data-driven methods in the food safety field.
Scope and approach: The major objective of this review is to introduce representative FRC databases with different focus areas, along with their chemical presentation, data quality and availability, and successful applications.
Key findings and conclusions: Over the past decades, over 50 FRC databases have been released, contributing significantly to scientific research, policymaking, and education. However, our analysis unveils persistent challenges such as delayed updates, accessibility concerns, reproducibility issues, suboptimal data quality, and inadequate coverage in underdeveloped regions. To address these shortcomings, we propose an initiative aimed at enhancing future FRC databases, prioritizing the principles of findability, accessibility, interoperability, and reusability. Additionally, we highlight the potential of future strategies, e.g., natural language processing, cheminformatics-enpowered suspect and non-targeted analysis, and genome mining, for the detection and analysis of emerging new FRCs outside of existing databases. By embracing these initiatives and strategies, we lay the groundwork for a robust framework facilitating enhanced food safety assessment and informed decision-making in the face of evolving challenges.

Non-target screening (NTS) methods are rapidly gaining in popularity, empowering researchers to search for an ever-increasing number of chemicals. Given this possibility, communicating the confidence of identification in an automated, concise and unambiguous manner is becoming increasingly important. In this study, we compiled several pieces of evidence necessary for communicating NTS identification confidence and developed a machine learning approach for classification of the identifications as reliable and unreliable. The machine learning approach was trained using data generated by four laboratories equipped with different instrumentation. The model discarded substances with insufficient identification evidence efficiently, while revealing the relevance of different parameters for identification. Based on these results, a harmonized IP-based system is proposed. This new NTS-oriented system is compatible with the currently widely used five level system. It increases the precision in reporting and the reproducibility of current approaches via the inclusion of evidence scores, while being suitable for automation.

Nontarget high-resolution mass spectrometry screening (NTS HRMS/MS) can detect thousands of organic substances in environmental samples. However, new strategies are needed to focus time-intensive identification efforts on features with the highest potential to cause adverse effects instead of the most abundant ones. To address this challenge, we developed MLinvitroTox, a machine learning framework that uses molecular fingerprints derived from fragmentation spectra (MS2) for a rapid classification of thousands of unidentified HRMS/MS features as toxic/nontoxic based on nearly 400 target-specific and over 100 cytotoxic endpoints from ToxCast/Tox21. Model development results demonstrated that using customized molecular fingerprints and models, over a quarter of toxic endpoints and the majority of the associated mechanistic targets could be accurately predicted with sensitivities exceeding 0.95. Notably, SIRIUS molecular fingerprints and xboost (Extreme Gradient Boosting) models with SMOTE (Synthetic Minority Oversampling Technique) for handling data imbalance were a universally successful and robust modeling configuration. Validation of MLinvitroTox on MassBank spectra showed that toxicity could be predicted from molecular fingerprints derived from MS2 with an average balanced accuracy of 0.75. By applying MLinvitroTox to environmental HRMS/MS data, we confirmed the experimental results obtained with target analysis and narrowed the analytical focus from tens of thousands of detected signals to 783 features linked to potential toxicity, including 109 spectral matches and 30 compounds with confirmed toxic activity.

Permanent rainbow trout (Oncorhynchus mykiss) cell lines represent potential in vitro alternatives to experiments with fish. We here developed a method to assess the bioaccumulation potential of anionic organic compounds in fish, using the rainbow trout liver-derived RTL-W1 cell line. Based on the availability of high quality in vivo bioconcentration (BCF) and biomagnification (BMF) data and the substances’ charge state at physiological pH, four anionic compounds were selected: pentachlorophenol (PCP), diclofenac (DCF), tecloftalam (TT) and benzotriazol-tert-butyl-hydroxyl-phenyl propanoic acid (BHPP). The fish cell line acute toxicity assay (OECD TG249) was used to derive effective concentrations 50 % and non-toxic exposure concentrations to determine exposure concentrations for bioaccumulation experiments. Bioaccumulation experiments were performed over 48 h with a total of six time points, at which cell, medium and plastic fractions were sampled and measured using high resolution tandem mass spectrometry after online solid phase extraction. Observed cell internal concentrations were over-predicted by K_OW-derived predictions while pH-dependent octanol–water partitioning (D_OW) and membrane lipid-water partitioning (D_MLW) gave better predictions of cell internal concentrations. Measured medium and cell internal concentrations at steady state were used to calculate RTL-W1-based BCF, which were compared to D_OW- or D_MLW-based model approaches and in vivo data. With the exception of PCP, the cell-derived BCF best compared to D_OW-based model predictions, which were higher than predictions based on D_MLW. All methods predicted the in vivo BCF for diclofenac well. For PCP, the cell-derived BCF was lowest although all BCF predictions underestimated the in vivo BCF by ≥ 1 order of magnitude. The RTL-W1 cells, and all other prediction methods, largely overestimated in vivo BMF, which were available for PCP, TT and BHPP. We conclude that the RTL-W1 cell line can supplement BCF predictions for anionic compounds. For BMF estimations, however, in vitro-in vivo extrapolations need adaptation or a multiple cell line approach.

Global change assessments have typically ignored synthetic chemical pollution, despite the rapid increase of pharmaceuticals, pesticides and industrial chemicals in the environment. Part of the problem reflects the multifarious origins of these micropollutants, which can derive from urban and agricultural sources. Understanding how micropollutants harm ecosystems is a major scientific challenge due to asymmetries of stress across trophic levels and ecological surprises generated by multiple drivers interacting in human-impacted landscapes. We used field assays above and below municipal wastewater treatment plants (WWTPs) in 60 sampling locations across 20 Swiss streams to test how micropollutants and nutrients originating from WWTPs affect two trophic levels (microbes and detritivores) and their role in leaf litter processing. Wastewater impacts were asymmetric across trophic levels, with the detritivore contribution declining relative to microbial-driven decomposition. The strength of negative impacts were context dependent, peaking at sites with the highest upstream abundances of detritivorous invertebrates. Diffuse pollution from intensive agriculture and wastewater-born micropollutants contributed to reduced litter processing rates, including indirect effects apparently mediated through negative influences of insecticides on detritivores. Asymmetries in stress responses across trophic levels can introduce quantitative changes in consumer–resource dynamics and leaf litter processing. This means functional redundancies at different trophic levels are insufficient to compensate for biodiversity losses, causing environmental stressors such as chemical pollutants to have pervasive ecosystem-level impacts.

Das Qualitätsziel einer 80%igen Elimination von Mikroverunreinigungen ist in der Schweiz für ausgewählte ARA auch bei Regenwetterverhältnissen stets einzuhalten. Hohe Niederschläge verursachen hohe Mischwasserzuläufe zur ARA, die in der Menge häufig das Doppelte der Trockenwettermenge übersteigt. Der Schmutzwasseranteil (Abwassermatrix) ist dann um das Vielfache verdünnt. Dieser Umstand verursacht hydraulische Effekte wie verkürzte hydraulische Aufenthaltszeiten in den Behandlungsstufen und führt zu verkürzten Kontakt- und Reaktionszeiten in den Reaktoren. U.a. können Kurzschlussströme entstehen, die die EMV erniedrigen können. Die EMVStufen in der Schweiz sind auf Mischwasserzulauf ausgelegt und es wurden über die letzten Jahre angepasste Betriebsstrategien für die verschiedenen
EMV-Verfahren erarbeitet, so dass in aller Regel das Qualitätsziel erreicht wird. Dennoch bleibt der sichere, wirtschaftliche sowie nachhaltige Betrieb der Anlagen anspruchsvoll mit hohen Herausforderungen.
Aktuelle Untersuchungen der Eawag zum Verhalten höher beladener GAK im Einzelfilter zeigten bei Kontakt mit stark verdünntem Abwasser trotz 20-minütiger Kontaktzeit verringerte Sorptionsleistung bis hin zur Desorption von Einzelsubstanzen. Ob dieser Umstand auch bei einem Parallelbetrieb zu beobachten ist, sollen weiterführende Untersuchungen im Praxisbetrieb von GAK-Filtrationen zeigen. Im Fall der GAK-Filtration der ARA Glarnerland mit langanhaltenden Regennachläufen und Schmelzwasserzulauf wurde eine PAK-Dosierung in die Biologie vorgeschaltet bzw. wird aktuell realisiert, um allenfalls Einbussen durch verdünnte Abwassermatrix abzufangen.

The distribution of microorganisms has long been assumed to be cosmopolitan and primarily controlled by the environment, but recent studies suggest that microbes may also exhibit strong biogeographical patterns driven by dispersal limitation. Past attempts to study the global biogeography of freshwater diatoms have always encountered the great difficulty of collecting taxonomically harmonized large-scale data. However, developments in molecular techniques and DNA metabarcoding provide a unique opportunity to overcome these limitations and to disclose diatom biodiversity at an unprecedented scale and resolution. Here, we assembled DNA metabarcoding data of freshwater benthic diatom communities sampled in seven geographic regions across the world to investigate how diatom diversity varies along latitude and to assess the proportion of genetic variants of these microorganisms which are exclusive or shared across regions. We observed significant differences in assemblages among climate zones and found that genetic richness is not affected by latitude, but by an island effect. The genetic resolution directly impacts the proportion of variants shared across regions; however, the majority of taxa remained specific to a single geographic region. Freshwater diatoms disperse over long distances and across oceans but at rates that allow the appearance of local genetic variants and the regionalization of assemblages. Future work should focus on putting these diversity dynamics into a temporal context, an approach that should be possible by bringing together new sequencing techniques and phylogeography.

An inadvertent consequence of pesticide use is aquatic pesticide pollution, which has prompted the implementation of mitigation measures in many countries. Water quality monitoring programs are an important tool to evaluate the efficacy of these mitigation measures. However, large interannual variability of pesticide losses makes it challenging to detect significant improvements in water quality and to attribute these improvements to the application of specific mitigation measures. Thus, there is a gap in the literature that informs researchers and authorities regarding the number of years of aquatic pesticide monitoring or the effect size (e.g., loss reduction) that is required to detect significant trends in water quality. Our research addresses this issue by combining two exceptional empirical data sets with modelling to explore the relationships between the achieved pesticide reduction levels due to mitigation measures and the length of the observation period for establishing statistically significant trends. Our study includes both a large (Rhine at Basel, ∼36,300 km²) and small catchment (Eschibach, 1.2 km²), which represent spatial scales at either end of the spectrum that would be realistic for monitoring programs designed to assess water quality. Our results highlight several requirements in a monitoring program to allow for trend detection. Firstly, sufficient baseline monitoring is required before implementing mitigation measures. Secondly, the availability of pesticide use data helps account for the interannual variability and temporal trends, but such data are usually lacking. Finally, the timing and magnitude of hydrological events relative to pesticide application can obscure the observable effects of mitigation measures (especially in small catchments). Our results indicate that a strong reduction (i.e., 70-90 %) is needed to detect a change within 10 years of monitoring data. The trade-off in applying a more sensitive method for change detection is that it may be more prone to false-positives. Our results suggest that it is important to consider the trade-off between the sensitivity of trend detection and the risk of false positives when selecting an appropriate method and that applying more than one method can provide more confidence in trend detection.

The Western Cape in South Africa has a Mediterranean climate, which has in part led to an abundance of agriculturally productive land supporting the wheat, deciduous fruit, wine, and citrus industries. South Africa is the leading pesticide user in Sub-Saharan Africa. There is limited data on the pesticide pollution of surface water over different seasons in low- and middle-income countries. We evaluated the seasonal drivers of aquatic pesticide pollution in three river catchments (Berg, Krom, and Hex Rivers) from July 2017 to June 2018 and April to July 2019, using 48 passive samplers. Our sampling followed the most severe drought (2015-2018) since recordings in 1960. Thus, our analyses focus on how drought and post-drought conditions may affect in-stream pesticide concentrations and loads. Samples were analyzed for 101 pesticide compounds using liquid chromatography - high-resolution mass spectrometry. Environmental Quality Standards (EQS) were used to assess the risks. We detected 60 pesticide compounds across the sampling periods. Our results indicate that all samples across all three catchments contained at least three pesticides and that the majority (83%) contained five or more pesticides. Approximately half the number of pesticides were detected after the drought in 2018. High concentration sums of pesticides (>1 μg/L) were detected over long time periods in the Hex River Valley (22 weeks) and in Piketberg (four weeks). Terbuthylazine, imidacloprid, and metsulfuron-methyl were detected in the highest concentrations, making up most of the detected mass, and were frequently above EQS. The occurrence of some pesticides in water generally correlated with their application and rainfall events. However, those of imidacloprid and terbuthylazine did not, suggesting that non-rainfall-driven transport processes are important drivers of aquatic pesticide pollution. The implementation of specific, scientifically sound, mitigation measures against aquatic pesticide pollution would require comprehensive pesticide application data as well as a targeted study identifying sources and transport processes for environmentally persistent pesticides.

The use of agrochemical and pharmaceutical active ingredients is essential in our modern society. Given the increased concern and awareness of the potential risks of some chemicals, there is a growing need to align with 'green chemistry' and 'safe and sustainable by design' principles and thus to evaluate the hazards of agrochemical and pharmaceutical active ingredients in early stages of R&D. We give an overview of the current challenges and opportunities to assess the principle of biodegradability in the environment. Development of new medium/high-throughput methodologies, combining predictive tools and wet-lab experimentation are essential to design biodegradable chemicals early in the active ingredient discovery and selection process.

Background Predicting in advance the behavior of new chemical compounds can support the design process of new products by directing the research toward the most promising candidates and ruling out others. Such predictive models can be data-driven using Machine Learning or based on researchers’ experience and depend on the collection of past results. In either case: models (or researchers) can only make reliable assumptions about compounds that are similar to what they have seen before. Therefore, consequent usage of these predictive models shapes the dataset and causes a continuous specialization shrinking the applicability domain of all trained models on this dataset in the future, and increasingly harming model-based exploration of the space.
Proposed solution In this paper, we propose cancels (CounterActiNg Compound spEciaLization biaS), a technique that helps to break the dataset specialization spiral. Aiming for a smooth distribution of the compounds in the dataset, we identify areas in the space that fall short and suggest additional experiments that help bridge the gap. Thereby, we generally improve the dataset quality in an entirely unsupervised manner and create awareness of potential flaws in the data. cancels does not aim to cover the entire compound space and hence retains a desirable degree of specialization to a specified research domain.
Results An extensive set of experiments on the use-case of biodegradation pathway prediction not only reveals that the bias spiral can indeed be observed but also that cancels produces meaningful results. Additionally, we demonstrate that mitigating the observed bias is crucial as it cannot only intervene with the continuous specialization process, but also significantly improves a predictor’s performance while reducing the number of required experiments. Overall, we believe that cancels can support researchers in their experimentation process to not only better understand their data and potential flaws, but also to grow the dataset in a sustainable way. All code is available under github.com/KatDost/Cancels .

A collaborative trial involving 16 participants from nine European countries was conducted within the NORMAN network in efforts to harmonise suspect and non-target screening of environmental contaminants in whole fish samples of bream (Abramis brama). Participants were provided with freeze-dried, homogenised fish samples from a contaminated and a reference site, extracts (spiked and non-spiked) and reference sample preparation protocols for liquid chromatography (LC) and gas chromatography (GC) coupled to high resolution mass spectrometry (HRMS). Participants extracted fish samples using their in-house sample preparation method and/or the protocol provided. Participants correctly identified 9-69 % of spiked compounds using LC-HRMS and 20-60 % of spiked compounds using GC-HRMS. From the contaminated site, suspect screening with participants’ own suspect lists led to putative identification of on average ∼145 and ∼20 unique features per participant using LC-HRMS and GC-HRMS, respectively, while non-target screening identified on average ∼42 and ∼56 unique features per participant using LC-HRMS and GC-HRMS, respectively. Within the same sub-group of sample preparation method, only a few features were identified by at least two participants in suspect screening (16 features using LC-HRMS, 0 features using GC-HRMS) and non-target screening (0 features using LC-HRMS, 2 features using GC-HRMS). The compounds identified had log octanol/water partition coefficient (K_OW) values from -9.9 to 16 and mass-to-charge ratios (m/z) of 68 to 761 (LC-HRMS and GC-HRMS). A significant linear trend was found between log K_OW and m/z for the GC-HRMS data. Overall, these findings indicate that differences in screening results are mainly due to the data analysis workflows used by different participants. Further work is needed to harmonise the results obtained when applying suspect and non-target screening approaches to environmental biota samples.

The assessment of persistence (P), bioaccumulation (B), and toxicity (T) of a chemical is a crucial first step at ensuring chemical safety and is a cornerstone of the European Union's chemicals regulation REACH (Registration, Evaluation, Authorization, and Restriction of Chemicals). Existing methods for PBT assessment are overly complex and cumbersome, have produced incorrect conclusions, and rely heavily on animal-intensive testing. We explore how new-approach methodologies (NAMs) can overcome the limitations of current PBT assessment. We propose two innovative hazard indicators, termed cumulative toxicity equivalents (CTE) and persistent toxicity equivalents (PTE). Together they are intended to replace existing PBT indicators and can also accommodate the emerging concept of PMT (where M stands for mobility). The proposed "toxicity equivalents" can be measured with high throughput in vitro bioassays. CTE refers to the toxic effects measured directly in any given sample, including single chemicals, substitution products, or mixtures. PTE is the equivalent measure of cumulative toxicity equivalents measured after simulated environmental degradation of the sample. With an appropriate panel of animal-free or alternative in vitro bioassays, CTE and PTE comprise key environmental and human health hazard indicators. CTE and PTE do not require analytical identification of transformation products and mixture components but instead prompt two key questions: is the chemical or mixture toxic, and is this toxicity persistent or can it be attenuated by environmental degradation? Taken together, the proposed hazard indicators CTE and PTE have the potential to integrate P, B/M and T assessment into one high-throughput experimental workflow that sidesteps the need for analytical measurements and will support the Chemicals Strategy for Sustainability of the European Union.

Aquatic pesticide pollution is an important issue worldwide. Countries rely on monitoring programs to observe water bodies quality and on models to evaluate pesticide risks for entire stream networks. Measurements are typically sparse and discontinuous which lead to issues in quantifying pesticide transport at the catchment scale. Therefore, it is essential to assess the performance of extrapolation approaches and provide guidance on how to extend monitoring programs to improve predictions. Here we present a feasibility study to predict pesticide levels in a spatially explicit manner in the Swiss stream network based on the national monitoring program quantifying organic micropollutants at 33 sites and spatially distributed explanatory variables. Firstly, we focused on a limited set of herbicides used on corn crops. We observed a significant relationship between herbicide concentrations and the areal fraction of hydrologically connected cornfields. Neglecting connectivity revealed no influence of areal corn coverage on the herbicide levels. Considering chemical properties of the compounds slightly improved the correlation. Secondly, we analysed a set of 18 pesticides widely used on different crops and monitored across the country. In this case, the areal fractions of arable or crop lands showed significant correlations with average pesticide concentrations. Similar results were found with average annual discharge or precipitation if two outlier sites were neglected. The correlations found in this paper explained only about 30 % of the observed variance leaving most of the variability unexplained. Accordingly, extrapolating the results from the existing monitoring sites to the Swiss river network comes with substantial uncertainty. Our study highlights possible reasons for weak matches, such as missing pesticide application data, limited set of compounds in the monitoring program, or a limited understanding of factors differentiating the loss rates from different catchments. Improving the data on pesticide applications will be essential to progress in this regard.

Combined sewer overflows (CSOs) are an important pathway of organic micropollutants from urban areas to open water bodies. Understanding the temporal dynamics of these micropollutants during overflow events is crucial for applying appropriate sampling methods and implementing effective management strategies. Yet, little is known about the dynamics of micropollutants in CSOs, because most studies report concentrations from single grab samples or event mean concentrations (EMCs). With unique high temporal resolution measurements (3 min), we show the real dynamics of polar organic micropollutants in CSOs of one small (2,700 people: P) and one large (159,000 P) urban catchment, for two micropollutant categories: (i) 33 micropollutants in municipal wastewater and (ii) 13 micropollutants from urban surface runoff. The concentration dynamics depend on the substance source and the catchment size. Indoor substances such as pharmaceuticals show high temporal dynamics with changes of 1 to 2 orders of magnitude within 9 min in the CSO of the small catchment. In contrast, outdoor substances at the small catchment and all substances at the large catchment display considerably lower variation. We tested various time-proportional sampling strategies to assess the range of error when estimating EMCs. We recommend an interval of 3 min to capture the dynamics of indoor substances in CSOs from small catchments. The results highlight that both future monitoring campaigns and the planning and management of urban wet-weather treatment systems will benefit from high temporal sampling resolutions, not only to understand dynamics but also to minimize errors of estimated EMCs.

The assessment of environmental hazard indicators such as persistence, mobility, toxicity, or bioaccumulation of chemicals often results in highly variable experimental outcomes. Persistence is particularly affected due to a multitude of influencing environmental factors, with biodegradation experiments resulting in half-lives spanning several orders of magnitude. Also, half-lives may lie beyond the limits of reliable half-life quantification, and the number of available data points per substance may vary considerably, requiring a statistically robust approach for the characterization of data. Here, we apply Bayesian inference to address these challenges and characterize the distributions of reported soil half-lives. Our model estimates the mean, standard deviation, and corresponding uncertainties from a set of reported half-lives experimentally obtained for a single substance. We apply our inference model to 893 pesticides and pesticide transformation products with experimental soil half-lives of varying data quantity and quality, and we infer the half-life distribution for each compound. By estimating average half-lives, their experimental variability, and the uncertainty of the estimations, we provide a reliable data source for building predictive models, which are urgently needed by regulatory authorities to manage existing chemicals and by industry to design benign, nonpersistent chemicals. Our approach can be readily adapted for other environmental hazard indicators.

Adsorption on activated carbon is a common process to remove pharmaceuticals in wastewater treatment. Activated carbon adsorption is usually applied to wastewater with a low content of biological degradable organics, i.e. after biological treatment. Especially low molecular weight (LMW) compounds are known to compete with pharmaceuticals for adsorption sites. The goal of this study was to test the hypothesis that biological treatment is necessary for efficient pharmaceutical removal. Source-separated urine after anaerobic storage (anaerobically stored urine) and after aerobic biological removal of organics without nitrification (organics-depleted urine) were used in this study. In anaerobically stored urine 60% of the organic compounds were LMW organics, of which about 40% were acetate and propionate. 74% of the DOC and 100% of acetate and propionate were removed during aerobic biological treatment. To investigate the effect of the organic compounds on pharmaceutical removal, sorption experiments with 19 spiked pharmaceuticals and one artificial sweetener were conducted with powdered activated carbon. Ethanol, another LMW organic, was included in the study, as it is regularly used for pharmaceutical spiking thereby strongly increasing the DOC content. The experiments showed that the adsorption of the pharmaceuticals and the sweetener were hardly affected by the easily biodegradable LMW organics or ethanol. Therefore, it was concluded that biological pre-treatment is not necessary for efficient pharmaceutical adsorption. Since acetate, propionate and ethanol contribute substantially to the DOC content but do not absorb UV light, the latter is recommended as indicator for pharmaceutical removal in solutions with high contents of biodegradable LMW organics.

Calls for supporting sustainability through more and better research rest on an incomplete understanding of scientific evidence use. We argue that a variety of barriers to a transformative impact of evidence arises from diverse actor motivations within different stages of evidence use. We abductively specify this variety in policy and practice arenas for three actor motivations (truth-seeking, sense-making, and utility-maximizing) and five stages (evidence production, uptake, influence on decisions, effects on sustainability outcomes, and feedback from outcome evaluations). Our interdisciplinary synthesis focuses on the sustainability challenge of reducing environmental and human health risks of agricultural pesticides. It identifies barriers resulting from (1) truth-seekers’ desire to reduce uncertainty that is complicated by evidence gaps, (2) sense-makers’ evidence needs that differ from the type of evidence available, and (3) utility-maximizers’ interests that guide strategic evidence use. We outline context-specific research–policy–practice measures to increase evidence use for sustainable transformation in pesticides and beyond.

Increasing production and use of chemicals and awareness of their impact on ecosystems and humans has led to large interest for broadening the knowledge on the chemical status of the environment and human health by suspect and non-target screening (NTS). To facilitate effective implementation of NTS in scientific, commercial and governmental laboratories, as well as acceptance by managers, regulators and risk assessors, more harmonisation in NTS is required. To address this, NORMAN Association members involved in NTS activities have prepared this guidance document, based on the current state of knowledge. The document is intended to provide guidance on performing high quality NTS studies and data interpretation while increasing awareness of the promise but also pitfalls and challenges associated with these techniques. Guidance is provided for all steps; from sampling and sample preparation to analysis by chromatography (liquid and gas - LC and GC) coupled via various ionisation techniques to high-resolution tandem mass spectrometry (HRMS/MS), through to data evaluation and reporting in the context of NTS. Although most experience within the NORMAN network still involves water analysis of polar compounds using LC–HRMS/MS, other matrices (sediment, soil, biota, dust, air) and instrumentation (GC, ion mobility) are covered, reflecting the rapid development and extension of the field. Due to the ongoing developments, the different questions addressed with NTS and manifold techniques in use, NORMAN members feel that no standard operation process can be provided at this stage. However, appropriate analytical methods, data processing techniques and databases commonly compiled in NTS workflows are introduced, their limitations are discussed and recommendations for different cases are provided. Proper quality assurance, quantification without reference standards and reporting results with clear confidence of identification assignment complete the guidance together with a glossary of definitions. The NORMAN community greatly supports the sharing of experiences and data via open science and hopes that this guideline supports this effort.

Assessing the persistence of organic micropollutants from field data has been notoriously laborious, requiring extensive data including emissions and chemical properties, and the application of detailed mass-balance models, which often contain parameters that are impossible to measure. To overcome some of these obstacles, we developed the concept of persistence benchmarking for large rivers that receive numerous emissions and provide enough residence time to observe the dissipation of compounds. We estimated the dissipation rate constants of 41 compounds (mostly active pharmaceutical ingredients) from five measurement campaigns in the Rhine and Danube rivers using concentration rate profiles with respect to carbamazepine. Dissipation rates clearly distinguished between known fast- and slow-degrading compounds, and campaign-specific boundary conditions had an influence on a minor subset of compounds only. Benchmarking provided reasonable estimates on summer total system half-lives in the Rhine compared to previous laboratory experiments and a mass-balance modeling study. Consequently, benchmarking can be a straightforward persistence assessment method of continuously emitted organic micropollutants in large river systems, especially when it is supported by field monitoring campaigns of proper analytical quality and spatial resolution.

Mitigation of undesired byproducts from ozonation of dissolved organic matter (DOM) such as aldehydes and ketones is currently hampered by limited knowledge of their precursors and formation pathways. Here, the stable oxygen isotope composition of H₂O₂ formed simultaneously with these byproducts was studied to determine if it can reveal this missing information. A newly developed procedure, which quantitatively transforms H₂O₂ to O₂ for subsequent ¹⁸O/¹⁶O ratio analysis, was used to determine the δ¹⁸O of H₂O₂ generated from ozonated model compounds (olefins and phenol, pH 3-8). A constant enrichment of ¹⁸O in H₂O₂ with a δ¹⁸O value of ∼59‰ implies that ¹⁶O-¹⁶O bonds are cleaved preferentially in the intermediate Criegee ozonide, which is commonly formed from olefins. H₂O₂ from the ozonation of acrylic acid and phenol at pH 7 resulted in lower ¹⁸O enrichment (δ¹⁸O = 47-49‰). For acrylic acid, enhancement of one of the two pathways followed by a carbonyl-H₂O₂ equilibrium was responsible for the smaller δ¹⁸O of H₂O₂. During phenol ozonation at pH 7, various competing reactions leading to H₂O₂ via an intermediate ozone adduct are hypothesized to cause lower δ¹⁸O in H₂O₂. These insights provide a first step toward supporting pH-dependent H₂O₂ precursor elucidation in DOM.

Ozonation of drinking water and wastewater is accompanied by the formation of disinfection byproducts (DBPs) such as low molecular weight aldehydes and ketones from the reactions of ozone with dissolved organic matter (DOM). By applying a recently developed non-target workflow, 178 carbonous and nitrogenous carbonyl compounds were detected during bench-scale ozonation of two lake waters and three secondary wastewater effluent samples and full-scale ozonation of secondary treated wastewater effluent. An overlapping subset of carbonyl compounds (20%) was detected in all water types. Moreover, wastewater effluents showed a significantly higher fraction of N-containing carbonyl compounds (30%) compared to lake water (17%). All carbonyl compounds can be classified in 5 main formation trends as a function of increasing specific ozone doses. Formation trends upon ozonation and comparison of results in presence and absence of the ^•OH radical scavenger DMSO in combination with kinetic and mechanistic information allowed to elucidate potential carbonyl structures. A link between the detected carbonyl compounds and their precursors was established by ozonating six model compounds (phenol, 4-ethylphenol, 4-methoxyphenol, sorbic acid, 3-buten-2-ol and acetylacetone). About one third of the detected carbonous carbonyl compounds detected in real waters was also detected by ozonating model compounds.
Evaluation of the non-target analysis data revealed the identity of 15 carbonyl compounds, including hydroxylated aldehydes and ketones (e.g. hydroxyacetone, confidence level (CL) = 1), unsaturated dicarbonyls (e.g. acrolein, CL = 1; 2-butene-1,4-dial, CL = 1; 4-oxobut-2-enoic acid, CL = 2) and also a nitrogen-containing carbonyl compound (2-oxo-propanamide, CL =1).
Overall, this study shows the formation of versatile carbonous and nitrogenous carbonyl compounds upon ozonation involving ozone and ^•OH reactions. Carbonyl compounds with unknown toxicity might be formed, and it could be demonstrated that acrolein, malondialdehyde, methyl glyoxal, 2-butene-1,4-dial and 4-oxo-pentenal are degraded during biological post-treatment.

Biodegradation holds promise as an effective and sustainable process for the removal of synthetic chemical pollutants. Nevertheless, rational engineering of biodegradation for pollutant remediation remains an unfulfilled goal, while chemical pollution of waters and soils continues to advance. Efforts to (i) identify functional bacteria from aquatic and soil microbiomes, (ii) assemble them into biodegrading consortia, and (iii) identify maintenance and performance determinants, are challenged by large number of pollutants and the complexity in the enzymology and ecology of pollutant biodegradation. To overcome these challenges, approaches that leverage knowledge from environmental bio-chem-informatics and metabolic engineering are crucial. Here, we propose a novel high-throughput bio-chem-informatics pipeline, to link chemicals and their predicted biotransformation pathways with potential enzymes and bacterial strains. Our framework systematically selects the most promising candidates for the degradation of chemicals with unknown biotransformation pathways and associated enzymes from the vast array of aquatic and soil bacteria. We substantiated our perspective by validating the pipeline for two chemicals with known or predicted pathways and show that our predicted strains are consistent with strains known to biotransform those chemicals. Such pipelines can be integrated with metabolic network analysis built upon genome-scale models and ecological principles to rationally design fit-for-purpose bacterial communities for augmenting deficient biotransformation functions and study operational and design parameters that influence their structure and function. We believe that research in this direction can pave the way for achieving our long-term goal of enhancing pollutant biodegradation.

Wastewater treatment plants (WWTPs) are a major source of micropollutants to surface waters. Currently, their chemical or biological monitoring is realized by using grab or composite samples, which provides only snapshots of the current wastewater composition. Especially in WWTPs with industrial input, the wastewater composition can be highly variable and a continuous assessment would be advantageous, but very labor and cost intensive. A promising concept are automated real-time biological early warning systems (BEWS), where living organisms are constantly exposed to the water and an alarm is triggered if the organism's responses exceed a harmful threshold of acute toxicity. Currently, BEWS are established for drinking water and surface water but are seldom applied to monitor wastewater. This study demonstrates that a battery of BEWS using algae (Chlorella vulgaris in the Algae Toximeter, bbe Moldaenke), water flea (Daphnia magna in the DaphTox II, bbe Moldaenke) and gammarids (Gammarus pulex in the Sensaguard, REMONDIS Aqua) can be adapted for wastewater surveillance. For continuous low-maintenance operation, a back-washable membrane filtration system is indispensable for adequate preparation of treated wastewater. Only minor deviations in the reaction of the organisms towards treated and filtered wastewater compared to surface waters were detected. After spiking treated wastewater with two concentrations of the model compounds diuron, chlorpyrifos methyl, and sertraline, the organisms in the different BEWS showed clear responses depending on the respective compound, concentration and mode of action. Immediate effects on photosynthetic activity of algae were detected for diuron exposure, and strong behavioral changes in water flea and gammarids after exposure to chlorpyrifos methyl or sertraline were observed, which triggered automated alarms. Different types of data analysis were applied to extract more information out of the specific behavioral traits, than only provided by the vendors algorithms. To investigate, whether behavioral movement changes can be linked to impact other endpoints, the effects on feeding activity of G. pulex were evaluated and results indicated significant differences between the exposures. Overall, these findings provide an important basis indicating that BEWS have the potential to act as alarm systems for pollution events in the wastewater sector.

Diatom species identification with DNA metabarcoding is an economical, fast and reliable alternative to identification via light microscopy for river quality monitoring. Using a short DNA sequence of the rbcL gene and 'Diat.barcode', a reference barcode library, enables the identification of more than 90% of the environmental sequences to species level in French rivers. But the completeness of this library is much lower in other regions, such as the tropical French overseas departments. A barcode library completion method using high-throughput sequencing data combined with microscopy count data from natural samples (Rimet et al. Citation2018) was applied and tested in rivers of Martinique and Guadeloupe (West Indies), for which only 45% of the environmental sequences could be identified to species level using Diat.barcode v9. Assigning barcodes to the most abundant species in the islands by this method is illustrated with Ulnaria goulardii and two new species belonging to Nupela and Epithemia, which are also described in this paper. The more complex situation of morphologically similar species is illustrated by reference to Gomphonema designatum and G. bourbonense. Using a combination of molecular and morphological data, their conspecificity, as G. bourbonense, is demonstrated with their reference barcodes. However, when several morphologically similar species and several environmental sequences belonging to the same clade are present, it is not possible to relate the barcodes to corresponding morphological species.
Applying this method enabled the Diat.barcode library (v.10) to be updated, with 84% of the environmental sequences from the West Indies now identifiable at the species level. However, many morphological species still lack barcodes. In these cases, more classical methods, such as cell isolation, Sanger sequencing and morphological observations of cultures, must be applied.

Human activity is a major driver of ecological and evolutionary change in wild populations and can have diverse effects on eukaryotic organisms as well as on environmental and host-associated microbial communities. Although host–microbiome interactions can be a major determinant of host fitness, few studies consider the joint responses of hosts and their microbiomes to anthropogenic changes. In freshwater ecosystems, wastewater is a widespread anthropogenic stressor that represents a multifarious environmental perturbation. Here, we experimentally tested the impact of treated wastewater on a keystone host (the freshwater isopod Asellus aquaticus) and its gut microbiome. We used a semi-natural flume experiment, in combination with 16S rRNA amplicon sequencing, to assess how different concentrations (0%, 30%, and 80%) of nonfiltered wastewater (i.e. with chemical toxicants, nutrients, organic particles, and microbes) versus ultrafiltered wastewater (i.e. only dissolved pollutants and nutrients) affected host survival, growth, and food consumption as well as mid- and hindgut bacterial community composition and diversity. Our results show that while host survival was not affected by the treatments, host growth increased and host feeding rate decreased with nonfiltered wastewater – potentially indicating that A. aquaticus fed on organic matter and microbes available in nonfiltered wastewater. Furthermore, even though the midgut microbiome (diversity and composition) was not affected by any of our treatments, nonfiltered wastewater influenced bacterial composition (but not diversity) in the hindgut. Ultrafiltered wastewater, on the other hand, affected both community composition and bacterial diversity in the hindgut, an effect that in our system differed between sexes. While the functional consequences of microbiome changes and their sex specificity are yet to be tested, our results indicate that different components of multifactorial stressors (i.e. different constituents of wastewater) can affect hosts and their microbiome in distinct (even opposing) manners and have a substantial impact on eco-evolutionary responses to anthropogenic stressors.

Graphitic carbon nitride (g-C₃N₄) nanomaterials hold great promise in diverse applications; however, their stability in engineering systems and transformation in nature are largely underexplored. We evaluated the stability, aging, and environmental impact of g-C₃N₄ nanosheets under the attack of free chlorine and reactive chlorine species (RCS), a widely used oxidant/disinfectant and a class of ubiquitous radical species, respectively. g-C₃N₄ nanosheets were slowly oxidized by free chlorine even at a high concentration of 200-1200 mg L^-1, but they decomposed rapidly when ClO· and/or Cl₂^•- were the key oxidants. Though Cl₂^•- and ClO· are considered weaker oxidants in previous studies due to their lower reduction potentials and slower reaction kinetics than ·OH and Cl·, our study highlighted that their electrophilic attack efficacy on g-C₃N₄ nanosheets was on par with ·OH and much higher than Cl·. A trace level of covalently bonded Cl (0.28-0.55 at%) was introduced to g-C₃N₄ nanosheets after free chlorine and RCS oxidation. Our study elucidates the environmental fate and transformation of g-C₃N₄ nanosheets, particularly under the oxidation of chlorine-containing species, and it also provides guidelines for designing reactive, robust, and safe nanomaterials for engineering applications.

Ozonation of natural waters is typically associated with the formation of carbonyl compounds (aldehydes, ketones and ketoacids), a main class of organic disinfection byproducts (DBPs). However, the detection of carbonyl compounds in water and wastewater is challenged by multiple difficulties inherent to their physicochemical properties. A non-target screening method involving the derivatisation of carbonyl compounds with p-toluenesulfonylhydrazine (TSH) followed by their analysis using liquid chromatography coupled to electrospray ionisation high-resolution mass spectrometry (LC-ESI-HRMS) and an advanced non-target screening and data processing workflow was developed. The workflow was applied to investigate the formation of carbonyl compounds during ozonation of different water types including lake water, aqueous solutions containing Suwannee River Fulvic acid (SRFA), and wastewater. A higher sensitivity for most target carbonyl compounds was achieved compared to previous derivatisation methods. Moreover, the method allowed the identification of known and unknown carbonyl compounds. 8 out of 17 target carbonyl compounds were consistently detected above limits of quantification (LOQs) in most ozonated samples. Generally, the concentrations of the 8 detected target compounds decreased in the order: formaldehyde > acetaldehyde > glyoxylic acid > pyruvic acid > glutaraldehyde > 2,3-butanedione > glyoxal > 1-acetyl-1-cyclohexene. The DOC concentration-normalised formation of carbonyl compounds during ozonation was higher in wastewater and SRFA-containing water than in lake water. The specific ozone doses and the type of the dissolved organic matter (DOM) played a predominant role for the extent of formation of carbonyl compounds. Five formation trends were distinguished for different carbonyl compounds. Some compounds were produced continuously upon ozonation even at high ozone doses, while others reached a maximum concentration at a certain ozone dose above which they decreased. Concentrations of target and peak areas of non-target carbonyl compounds during full-scale ozonation at a wastewater treatment plant showed an increase as a function of the specific ozone dose (sum of 8 target compounds ∼ 280 µg/L at 1 mgO₃/mgC), followed by a significant decrease after biological sand filtration (> 64-94% abatement for the different compounds). This highlights the biodegradability of target and non-target carbonyl compounds and the importance of biological post-treatment.

Convolutional Neural Networks (CNNs) offer an alternative to the image cross-correlation methods used in Particle Image Velocimetry (PIV) to reconstruct the fluid velocity field from the experimental recording. Despite the flexibility of CNNs, the accuracy and robustness of the standard image processing remains unsurpassed for general PIV data. As CNNs are non-linear and typically entail up to millions of trainable parameters, they require large and carefully designed training datasets to avoid over-fitting and to obtain results that are accurate for a wide range of flow conditions and length scales. Most training datasets consist of PIV-like data that are generated from displacement fields resulting from numerical flow simulations, which, in addition of being computationally expensive, may be able to inform the network only about relatively few classes of flow problems. To overcome this issue and improve the accuracy of the velocity reconstructed by CNNs, we propose to train the networks with synthetic PIV-like data generated from random displacement fields. The underlying idea is that the training dataset simply needs to teach the network about the kinematic relationship between position and velocity. These kinematic training datasets are computationally inexpensive and may allow a much richer variability in terms of length scales by varying the generation parameters. By training a state-of-the-art CNN, we investigate the accuracy of the reconstructed displacement and velocity with synthetic and experimental test cases, such as a sinusoidal flow and wind-tunnel data from a turbulent-boundary-layer and a cylinder-wake experiment. We demonstrate that kinematic training can drastically improve the accuracy of the CNN and allows the network to outperform conventional cross-correlation methods, being more robust with respect to data noise and providing reconstructed velocity fields that have considerably higher spatial resolution (at pixel level).

In order to protect European Union (EU) drinking water resources from chemical contamination, criteria for identifying persistent, mobile, and toxic (PMT) chemicals and very persistent and very mobile (vPvM) chemicals under the EU REACH Regulation were proposed by the German Environment Agency (Umweltbundesamt—UBA). Additionally, new hazard classes for PMT and vPvM substances in the revised EU classification, labeling, and packaging (CLP Regulation) are intended. Therefore, a reliable approach in the identification of potential drinking water resource contaminants is needed. The scientific basis of the property-based PMT/vPvM criteria, focusing on mobility, which dictates the migration of chemical drinking water sources, was evaluated, and a critical analysis of the deviation of sorption metrics from simple behavior was carried out. Based on our evaluation, a K_oc may be used for nonionic substances on a screening level only, requiring a higher tier assessment. It is considered inappropriate for hydrophilic and ionizable chemicals, particularly for soils with low organic carbon contents. The nonextractable residue formation is complex and not well understood but remains significant in limiting the mobility of chemicals through soils and sediments. In order to inform the EU commission's work on the introduction of new hazard classes for PMT and vPvM substances into the European legislation, the derivation of a tiered approach is proposed, which utilizes the weight of evidence available, with adoption of appropriate higher tier models commensurate with the nature of the substance and the data available.

Delayed toxicity is a phenomenon observed for aquatic invertebrates exposed to nicotinic acetylcholine receptor (nAChR) agonists, such as neonicotinoids. Furthermore, recent studies have described an incomplete elimination of neonicotinoids by exposed amphipods. However, a mechanistic link between receptor binding and toxicokinetic modeling has not been demonstrated yet. The elimination of the neonicotinoid thiacloprid in the freshwater amphipod Gammarus pulex was studied in several toxicokinetic exposure experiments, complemented with in vitro and in vivo receptor-binding assays. Based on the results, a two-compartment model was developed to predict the uptake and elimination kinetics of thiacloprid in G. pulex. An incomplete elimination of thiacloprid, independent of elimination phase duration, exposure concentrations, and pulses, was observed. Additionally, the receptor-binding assays indicated irreversible binding of thiacloprid to the nAChRs. Accordingly, a toxicokinetic-receptor model consisting of a structural and a membrane protein (including nAChRs) compartment was developed. The model successfully predicted internal thiacloprid concentrations across various experiments. Our results help in understanding the delayed toxic and receptor-mediated effects toward arthropods caused by neonicotinoids. Furthermore, the results suggest that more awareness toward long-term toxic effects of irreversible receptor binding is needed in a regulatory context. The developed model supports the future toxicokinetic assessment of receptor-binding contaminants.

The application of mass spectrometry imaging (MSI) is a promising tool to analyze the spatial distribution of organic contaminants in organisms and thereby improve the understanding of toxicokinetic and toxicodynamic processes. MSI is a common method in medical research but has been rarely applied in environmental science. In the present study, the suitability of MSI to assess the spatial distribution of organic contaminants and their biotransformation products (BTPs) in the aquatic invertebrate key species Gammarus pulex was studied. Gammarids were exposed to a mixture of common organic contaminants (carbamazepine, citalopram, cyprodinil, efavirenz, fluopyram and terbutryn). The distribution of the parent compounds and their BTPs in the organisms was analyzed by two MSI methods (MALDI- and DESI-HRMSI) after cryo-sectioning, and by LC-HRMS/MS after dissection into different organ compartments. The spatial distribution of contaminats in gammarid tissue could be successfully analyzed by the different analytical methods. The intestinal system was identified as the main site of biotransformation, possibly due to the presence of biotransforming enzymes. LC-HRMS/MS was more sensitive and provided higher confidence in BTP identification due to chromatographic separation and MS/MS. DESI was found to be the more sensitive MSI method for the analyzed contaminants, whereas additional biomarkers were found using MALDI. The results demonstrate the suitability of MSI for investigations on the spatial distribution of accumulated organic contaminants. However, both MSI methods required high exposure concentrations. Further improvements of ionization methods would be needed to address environmentally relevant concentrations.

Bioaccumulation of organic contaminants from contaminated food sources might pose an underestimated risk toward shredding invertebrates. This assumption is substantiated by monitoring studies observing discrepancies of predicted tissue concentrations determined from laboratory-based experiments compared with measured concentrations of systemic pesticides in gammarids. To elucidate the role of dietary uptake in bioaccumulation, gammarids were exposed to leaf material from trees treated with a systemic fungicide mixture (azoxystrobin, cyprodinil, fluopyram, and tebuconazole), simulating leaves entering surface waters in autumn. Leaf concentrations, spatial distribution, and leaching behavior of fungicides were characterized using liquid chromatography coupled with high-resolution tandem mass spectrometry (LC-HRMS/MS) and matrix-assisted laser desorption ionization-mass spectrometric imaging. The contribution of leached fungicides and fungicides taken up from feeding was assessed by assembling caged (no access) and uncaged (access to leaves) gammarids. The fungicide dynamics in the test system were analyzed using LC-HRMS/MS and toxicokinetic modeling. In addition, a summer scenario was simulated where water was the initial source of contamination and leaves contaminated by sorption. The uptake, translocation, and biotransformation of systemic fungicides by trees were compound-dependent. Internal fungicide concentrations of gammarids with access to leaves were much higher than in caged gammarids of the autumn scenario, but the difference was minimal in the summer scenario. In food choice and dissectioning experiments gammarids did not avoid contaminated leaves and efficiently assimilated contaminants from leaves, indicating the relevance of this exposure pathway in the field. The present study demonstrates the potential impact of dietary uptake on in situ bioaccumulation for shredders in autumn, outside the main application period. The toxicokinetic parameters obtained facilitate modeling of environmental exposure scenarios. The uncovered significance of dietary uptake for detritivores warrants further consideration from scientific as well as regulatory perspectives.

The acceleration of global climate change draws increasing attention towards interactive effects of temperature and organic contaminants. Many studies reported a higher sensitivity of aquatic invertebrates towards contaminant exposure with increasing or fluctuating temperatures. The hypothesis of this study was that the higher sensitivity of invertebrates is associated with the changes of toxicokinetic processes that determine internal concentrations of contaminants and consequently toxic effects. Therefore, the influence of temperature on toxicokinetic processes and the underlying mechanisms were studied in two key amphipod species (Gammarus pulex and Hyalella azteca). Bioconcentration experiments were carried out at four different temperatures with a mixture of 12 exposure relevant polar organic contaminants. Tissue and medium samples were taken in regular intervals and analysed by online solid-phase extraction liquid chromatography high-resolution tandem mass spectrometry. Subsequently, toxicokinetic rates were modelled and analysed in dependence of the exposure temperature using the Arrhenius equation. An exponential relationship between toxicokinetic rates versus temperature was observed and could be well depicted by applying the Arrhenius equation. Due to a similar Arrhenius temperature of uptake and elimination rates, the bioconcentration factors of the contaminants were generally constant across the temperature range. Furthermore, the Arrhenius temperature of the toxicokinetic rates and respiration was mostly similar. However, in some cases (citalopram, cyprodinil), the bioconcentration factor appeared to be temperature dependent, which could potentially be explained by the influence of temperature on active uptake mechanisms or biotransformation. The observed temperature effects on toxicokinetics may be particularly relevant in non-equilibrated systems, such as exposure peaks in summer as exemplified by the exposure modelling of a field measured pesticide peak where the internal concentrations increased by up to fourfold along the temperature gradient. The results provide novel insights into the mechanisms of chemical uptake, biotransformation and elimination in different climate scenarios and can improve environmental risk assessment.

Ferrate (Fe(VI)) is a novel oxidant that can be used to mitigate disinfection byproduct (DBP) precursors. However, the reaction of Fe(VI) with organic nitrogen, which is a potential precursor of potent nitrogenous DBPs, remains largely unexplored. The present work aimed to identify the kinetics and products for the reaction of Fe(VI) with primary amines, notably amino acids. A new kinetic model involving ionizable intermediates was proposed and can describe the unusual pH effect on the Fe(VI) reactivity toward primary amines and amino acids. The Fe(VI) oxidation of phenylalanine produced a mixture of nitrile, nitrite/nitrate, amide, and ammonia, while nitroalkane was an additional product in the case of glycine. The product distribution for amino acids significantly differed from that of uncarboxylated primary amines that mainly generate nitriles. A general reaction pathway for primary amines and amino acids was proposed and notably involved the formation of imines, the degradation of which was affected by the presence of a carboxylic group. In comparison, ozonation led to higher yields of nitroalkanes that could be readily converted to potent halonitroalkanes during chlor(am)ination. Based on this study, Fe(VI) can effectively mitigate primary amine-based, nitrogenous DBP precursors with little formation of toxic halonitroalkanes.

Micropollutants have become a serious environmental problem by threatening ecosystems and the quality of drinking water. This account investigates if advanced AI can be used to find solutions for this problem. We review background, the challenges involved, and the current state-of-the-art of quantitative structure–bio-degradation relationships (QSBR). We report on recent progress combining experiment, quantum chemistry (QC) and chemoinformatics, and provide a perspective on potential future uses of AI technology to help improve water quality.

Here, we report six novel, easily accessible BODIPY-based agents for cancer treatment. In contrast to established photodynamic therapy (PDT) agents, these BODIPY-based compounds show additional photothermal activity and their cytotoxicity is not dependent on the generation of reactive oxygen species (ROS). The agents show high photocytotoxicity upon irradiation with light and low dark toxicity in different cancer cell lines in 2D culture as well as in 3D multicellular tumor spheroids (MCTSs). The ratio of dark to light toxicity (phototoxic index, PI) of these agents reaches striking values exceeding 830,000 after irradiation with energetically low doses of light at 630 nm. The oxygen-dependent mechanism of action (MOA) of established photosensitizers (PSs) hampers effective clinical deployment of these agents. Under hypoxic conditions (0.2% O₂), which are known to limit the efficiency of conventional PSs in solid tumors, photocytotoxicity was induced at the same concentration levels, indicating an oxygen-independent photothermal MOA. With a PI exceeding 360,000 under hypoxic conditions, both PI values are the highest reported to date. We anticipate that small molecule agents with a photothermal MOA, such as the BODIPY-based compounds reported in this work, may overcome this barrier and provide a new avenue to cancer therapy.

Persistent, mobile and toxic (PMT), and very persistent and very mobile (vPvM) substances pose a threat to the water cycle but are often not covered in conventional environmental monitoring programs. Within this realm of substances, one compound class of concern are pesticides and their transformation products as they are deliberately introduced into the environment. To detect very polar anionic substances, including many pesticide transformation products with log D_OW values ranging between -7.4 and 2.2, an ion chromatography high-resolution mass spectrometry method was developed in this study. Since inorganic anions, such as chloride and sulfate, interfere with the analysis of organic species, their removal via precipitation with Ba/Ag/H cartridges was assessed. To improve LOQs, vacuum-assisted evaporative concentration (VEC) was evaluated. By using VEC and removing inorganic salt ions, the median LOQ improved from 100 ng/L in evian® water without sample treatment to 10 ng/L after enrichment and 30 ng/L in karst groundwater. Using this method, twelve out of 64 substances covered by the final method were found in karst groundwater in concentrations of up to 5600 ng/L, and seven exceeded 100 ng/L. To the authors' knowledge, the dimethenamid TP M31 and chlorothalonil TP SYN548008 were detected for the first time in groundwater samples. The coupling to a high-resolution mass spectrometer also allows for non-target screening and hence, this method presents a powerful tool to tackle PMT/vPvM substances.

Identifying a chemical's potential for biotransformation in the aquatic environment is crucial to predict its fate and manage its potential hazards. Due to the complexity of natural water bodies, especially river networks, biotransformation is often studied in laboratory experiments, assuming that study outcomes can be extrapolated to compound behavior in the field. Here, we investigated to what extent outcomes of laboratory simulation studies indeed reflect biotransformation kinetics observed in riverine systems. To determine in-field biotransformation, we measured loads of 27 wastewater treatment plant effluent-borne compounds along the Rhine and its major tributaries during two seasons. Up to 21 compounds were detected at each sampling location. Measured compound loads were used in an inverse model framework of the Rhine river basin to derive k'_bio,field values – a compound-specific parameter describing the compounds' average biotransformation potential during the field studies. To support model calibration, we performed phototransformation and sorption experiments with all the study compounds, identifying 5 compounds that are susceptible towards direct phototransformation and determining K_oc values covering four orders of magnitude. On the laboratory side, we used a similar inverse model framework to derive k'_bio,lab values from water-sediment experiments run according to a modified OECD 308-type protocol. The comparison of k'_bio,lab and k'_bio,field revealed that their absolute values differed, pointing towards faster transformation in the Rhine river basin. Yet, we could demonstrate that relative rankings of biotransformation potential and groups of compounds with low, moderate and high persistence agree reasonably well between laboratory and field outcomes. Overall, our results provide evidence that laboratory-based biotransformation studies using the modified OECD 308 protocol and k'_bio values derived thereof bear considerable potential to reflect biotransformation of micropollutants in one of the largest European river basins.

The environmental relevance of standard biodegradation tests such as OECD 309 has been questioned. Challenges include the interpretation of changing degradation kinetics over the 60–90 incubation days and the effects of chemical spiking on the microbial community. To ameliorate these weaknesses, we evaluated a modified OECD 309 test using water and sediment from three Swedish rivers. For each river, we had three treatments (no spiking, 0.5 μg L^–1 spiking, and 5 μg L^–1 spiking). The dissipation of a mixture of 56–80 spiked chemicals was followed over 14 days. Changes in dissipation kinetics during the incubation were interpreted as a departure of the microbial community from its initial (natural) state. The biodegradation kinetics were first-order throughout the incubation in the no spiking and 0.5 μg L^–1 spiking treatments for almost all chemicals, but for the 5 μg L^–1 treatment, more chemicals showed changes in kinetics. The rate constants in the no spiking and 0.5 μg L^–1 treatments agreed within a factor of 2 for 35 of 37 cases. We conclude that the environmental relevance of OECD 309 is improved by considering only the initial biodegradation phase and that it is not compromised by spiking multiple chemicals at 0.5 μg L^–1.

While man-made chemicals in the environment are ubiquitous and a potential threat to human health and ecosystem integrity, the environmental fate of chemical contaminants such as pharmaceuticals is often poorly understood. Biodegradation processes driven by microbial communities convert chemicals into transformation products (TPs) that may themselves have adverse ecological effects. The detection of TPs formed during biodegradation has been continuously improved thanks to the development of TP prediction algorithms and analytical workflows. Here, we contribute to this advance by (i) reviewing past applications of TP identification workflows, (ii) applying an updated workflow for TP prediction to 42 pharmaceuticals in biodegradation experiments with activated sludge, and (iii) benchmarking 5 different pathway prediction models, comprising 4 prediction models trained on different datasets provided by enviPath, and the state-of-the-art EAWAG pathway prediction system. Using the updated workflow, we could tentatively identify 79 transformation products for 31 pharmaceutical compounds. Compared to previous works, we have further automatized several steps that were previously performed by hand. By benchmarking the enviPath prediction system on experimental data, we demonstrate the usefulness of the pathway prediction tool to generate suspect lists for screening, and we propose new avenues to improve their accuracy. Moreover, we provide a well-documented workflow that can be (i) readily applied to detect transformation products in activated sludge and (ii) potentially extended to other environmental studies.

Direct and indirect threats by organic micropollutants can only be reliably assessed and prevented if the exposure to these chemicals is known, which in turn requires a confident estimate of their emitted amounts into the environment. APIs (Active Pharmaceutical Ingredients) enter surface waters mostly through the sewer system and wastewater treatment plants (WWTPs). However, their effluent fluxes are highly variable and influenced by several different factors that challenge robust emission estimates. Here, we defined a dimensionless, theoretically consumption-independent ‘escape factor’ (k_esc) for estimating the amount of APIs (expected to be) present in WWTP effluents. The factor is determined as the proportion of marketed and actually emitted amounts of APIs. A large collection of German and Swiss monitoring datasets were analyzed to calculate stochastic k_esc values for 31 APIs, reflecting both the magnitude and uncertainty of consumption-normalised emissions. Escape factors provide an easy-to-use tool for the estimation of average API emissions and expected variability from numerous WWTPs given that consumption data are provided, thereby supporting simulation modeling of the fate of APIs in stream networks or exposure assessments.

Pesticides are used throughout the tropics in large amounts to protect crops against pests and weeds. These chemicals may be lost to the aquatic environment and impair its ecological status and the water quality for human consumption. Despite intensive use of pesticides in many developing countries, the knowledge of pesticide levels in aquatic ecosystems is often very limited. In this study, we try to fill this gap for an horticultural catchment of 35.9 km² in Costa Rica, which is intensively used by small-holder farming for potatoes and vegetable production. We continuously monitored pesticides in the Tapezco river during two consecutive rainy seasons with passive sampling systems and screened for a broad set of polar and non-polar pesticides. Spatially distributed measurements revealed high pesticide concentrations of many fungicides, herbicides and insecticides throughout the watershed. Concentration ranges revealed little spatio-temporal variation. From an ecotoxicological point of view, the insecticide levels - notably chlorpyrifos and cypermethrin - were most critical. The observed concentration levels exceeded chronic environmental quality standards more than 100-fold at all sites. These high insecticide levels were partially reflected in the community composition of the macroinvertebrates. Available data revealed a poor status at two upstream locations according to the Costa Rican Biological Monitoring Working Party (BMWP-CR) Index and the SPEAR_pesticides index. However, the indices indicated a good quality at most downstream sites despite their high pesticide levels. The wide-spread occurrence of high pesticide levels demonstrated that the relevant sources and pathways existed throughout the catchment. Field observations and survey data showed the relevance of point sources due to poor pesticide handling as well as diffuse losses from fields, which are strongly enhanced by the steep terrain and linear structures like gullies connecting fields with the stream. Mitigation measures to reduce pesticide losses have to account for these different source-flowpath combinations.

Motivation: Transformation products (TPs) of man-made chemicals, formed through microbially mediated transformation in the environment, can have serious adverse environmental effects, yet the analytical identification of TPs is challenging. Rule-based prediction tools are successful in predicting TPs, especially in environmental chemistry applications that typically have to rely on small datasets, by imparting the existing knowledge on enzyme-mediated biotransformation reactions. However, the rules extracted from biotransformation reaction databases usually face the issue of being over/under-generalized and are not flexible to be updated with new reactions.
Results: We developed an automatic rule extraction tool called enviRule. It clusters biotransformation reactions into different groups based on the similarities of reaction fingerprints, and then automatically extracts and generalizes rules for each reaction group in SMARTS format. It optimizes the genericity of automatic rules against the downstream TP prediction task. Models trained with automatic rules outperformed the models trained with manually curated rules by 30% in the area under curve (AUC) scores. Moreover, automatic rules can be easily updated with new reactions, highlighting enviRule's strengths for both automatic extraction of optimized reactions rules and automated updating thereof.
Availability and implementation: enviRule code is freely available at https://github.com/zhangky12/enviRule.

Cyanobacterial blooms affect aquatic ecosystems across the globe and one major concern relates to their toxins such as microcystins (MC). Yet, the ecotoxicological risks, particularly non-lethal effects, associated with other co-produced secondary metabolites remain mostly unknown. Here, we assessed survival, morphological alterations, swimming behaviour and cardiovascular functions of zebrafish (Danio rerio) upon exposure to cyanobacterial extracts of two Brazilian Microcystis strains. We verified that only MIRS-04 produced MCs and identified other co-produced cyanopeptides also for the MC non-producer NPCD-01 by LC-HRMS/MS analysis. Both cyanobacterial extracts, from the MC-producer and non-producer, caused acute toxicity in zebrafish with LC₅₀ values of 0.49 and 0.98 mg_{dw_biomass}/mL, respectively. After exposure to MC-producer extract, additional decreased locomotor activity was observed. The cyanopeptolin (micropeptin K139) contributed 52% of the overall mortality and caused oedemas of the pericardial region. Oedemas of the pericardial area and prevented hatching were also observed upon exposure to the fraction with high abundance of a microginin (Nostoginin BN741) in the extract of the MC non-producer. Our results further add to the yet sparse understanding of lethal and sublethal effects caused by cyanobacterial metabolites other than MCs and the need to better understand the underlying mechanisms of the toxicity. We emphasize the importance of considering mixture toxicity of co-produced metabolites in the ecotoxicological risk assessment of cyanobacterial bloom events, given the importance for predicting adverse outcomes in fish and other organisms.

Marine biofouling is a natural process by which many organisms colonize and grow in submerged structures, causing serious economic consequences for the maritime industry. Geniculate calcareous algae (GCA; Corallinales, Rhodophyta) produce bioactive secondary metabolites and are a promise for new antifouling compounds. Here, we investigated the antifouling activity of four GCA species—Amphiroa beauvoisii, Jania sagittata (formerly Cheilosporum sagittatum), Jania crassa, and Jania prolifera (formerly Amphiroa flabellata)—from the Brazilian coast against macro- and microorganisms. Simultaneously, metabolomic tools were applied to assess the chemical profiles of these seaweeds using gas chromatography coupled to mass spectrometry (GC-MS). Data analysis by principal component and molecular networking analyses used the global natural products social molecular networking platform (GNPS). Our results showed that all extracts were active against different strains of marine bacteria and that the J. sagittata (JsSI) extract showed the highest percentage of bacterial inhibition. The J. sagittata (JsSI) extract was the most active against the mussel Perna perna, showing 100% byssus inhibition. Regarding toxicity, only the J. crassa (JcP) extract showed a 20% mortality rate. The chemical profiles of the evaluated GCA extracts differed qualitatively and quantitatively. Yet, the steroid (3β)-cholest-5-en-3-ol was the major compound commonly identified in all extracts, with the exception of J. sagittata (JsSI). Moreover, we observed intra- and interspecific chemical variabilities among GCA extracts for the different populations, which could explain their antifouling activity variability. This study contributed new information about the chemical compounds produced by this group of seaweeds and showed its antifouling potential. These GCA species may be the subject of future studies to obtain new bioactive compounds with biotechnological potential in maritime areas.

Plant Protection Products (PPPs) pose a threat to surface water quality worldwide. While small streams compose the majority of the stream lengths and are crucial for biodiversity, their exposure patterns to PPPs and transformation products (TPs) are largely understudied in dry periods. This knowledge gap can lead to ineffective monitoring strategies for addressing water quality issues. Here, we focus on two extended dry periods the in-depth analysis of a unique continuous high-frequency (20 min) concentrations dataset for 60 PPPs and TPs. The dataset refers to the monitoring of a small tile-drained agricultural stream over 41 days from May to July in 2019. The overall 2560 concentration data per compound obtained with the on-site mass spectrometer MS²Field platform revealed: (i) surprisingly high maximum concentrations (hundreds to thousands ng/l for some compounds) over extended periods of time, (ii) novel diel fluctuations of concentrations in the order of hundreds of ng/l for some PPPs and TPs, (iii) unexpected high concentrations (up to 220 ng/l) of a legacy compound (the fungicide oxadixyl withdrawn from the Swiss market in 2005). We hypothesized the cause of our findings was rooted in high PPPs levels in the shallow groundwater. To investigate this, we complemented our measurements with the long-term Swiss national monitoring program integrating samples over 14 days at the same location. The continuous long-term measurements found a few PPPs all year-round, thus indicating the presence of persistent contamination sources in the catchment. Next, we collected spatially distributed grab samples in tile drain outlets and stream water on a dry summer day in 2020. The dry-day campaign not only confirmed our hypothesis given the measured high concentrations of PPPs and TPs in tile drain outlets but also highlighted large spatial variability in measured concentrations along the stream. Hereafter, we highlight the questions that different monitoring schemes can answer in dry conditions. This information was relevant to observe for the first time, and thus foresee, the dynamic patterns of PPPs and TPs in the aquatic ecosystem in dry summer conditions, with the latter generally becoming more frequent due to climate change.

Protected agriculture boosts the production of vegetables, berries and fruits, and it plays a pivotal role in guaranteeing food security globally in the face of climate change. Remote sensing is proven to be useful for identifying the presence of (low-tech) plastic greenhouses and plastic mulches. However, the classification accuracy notoriously decreases in the presence of small-scale farming, heterogeneous land cover and unaccounted seasonal management of protected agriculture. Here, we present the random forest-based pixel-level Open field and Protected Agriculture land cover Classifier (OPAC) developed using Sentinel-2 L2A data. OPAC is trained using tiles from Switzerland over 2 years and the Almeria region in Spain over 1 acquisition day. OPAC classifies eight land covers typical of open field and protected agriculture (plastic mulches, low-tech greenhouses and for the first time high-tech greenhouses). Finally, we assess (1) how the land covers in OPAC are labelled in the Sentinel-2 Scene Classification Layer (SCL) and (2) the correspondence between pixels classified as protected agriculture by OPAC and by the best performing Advanced Plastic Greenhouse Index (APGI). To reduce anthropogenic land covers, we constrain the classification task to agricultural areas retrieved from cadastral data or the Corine Land Cover map. The 5-fold cross-validation reveals an overall accuracy of 92% but other classification scores are moderate when keeping the separation among the three classes of protected agriculture. However, all scores substantially improve upon grouping the three classes into one (with an Intersection Over Union of 0.58 as an average among the scores of the three classes and of 0.98 for one single class). Given the recently acknowledged importance of Sentinel-2 Band 1 (central wavelength of 443 nm), the classification accuracy of OPAC for the Swiss small-scale farming is mostly limited by the band's reduced spatial accuracy (60 m). A careful visual assessment indicates that OPAC achieves satisfactory generalization capabilities also in North European (the Netherlands) and four Mediterranean areas (Spain, Italy, Crete and Turkey) without the need of adding location and temporal specific information. There is good agreement among natural land covers classified by OPAC and the SCL. However, the SCL does not have a class for protected agriculture, the latter being often classified as clouds. APGI achieved similar to lower classification accuracies than OPAC. Importantly, the APGI classification task depends on a user-defined space- and time-specific threshold, whereas OPAC does not. Therefore, OPAC paves the way for rapid mapping of protected agriculture at continental scale.

Our study uses regional-scale maps to quantify carbon storage and sequestration from different land use types to evaluate the effects of future land use scenarios. We developed an integrated modeling approach to assess the spatiotemporal impacts of land use/cover change (LUCC) on the provision and value of the carbon storage and sequestration during the historical period (2000-2019) and predicted scenarios (2019-2046) in the Jiroft plain, Iran. We integrated several analytic tools for our analysis, which was comprised of Google Earth Engine (GEE), Cellular Automata Markov Chain (CA-MC) model, Intensity Analysis (IAA), and the Integrated Valuation of Ecosystem Services and Tradeoffs (InVEST) model. Our results demonstrate that: (1) agriculture and urban expansion led to a considerable decrease in carbon storage, mainly due to rapid deforestation from 2000-2019; (2) if the historical trend continues under the business as usual (BAU) scenario, it will lead to considerable social costs due to the loss of stored carbon in the plain (2,624,113 Mg) with an annual average sequestration loss of -475,547 Mg; (3) the downward carbon sequestration trend could potentially be reversed by employing the environmentally sound planning (ESP) scenario that is estimated to save 3,705,491 Mg in carbon storage, with annual average sequestration gain of + 605,830 Mg. The design scenarios provide a useful guide for policymakers and local governments to help understand the potential outcomes of the various development strategies, which will ultimately lead to more effective ecosystem management.

Anaerobic microbial respiration in suboxic and anoxic environments often involves particulate ferric iron (oxyhydr-)oxides as terminal electron acceptors. To ensure efficient respiration, a widespread strategy among iron-reducing microorganisms is the use of extracellular electron shuttles (EES) that transfer two electrons from the microbial cell to the iron oxide surface. Yet, a fundamental understanding of how EES-oxide redox thermodynamics affect rates of iron oxide reduction remains elusive. Attempts to rationalize these rates for different EES, solution pH, and iron oxides on the basis of the underlying reaction free energy of the two-electron transfer were unsuccessful. Here, we demonstrate that broadly varying reduction rates determined in this work for different iron oxides and EES at varying solution chemistry as well as previously published data can be reconciled when these rates are instead related to the free energy of the less exergonic (or even endergonic) first of the two electron transfers from the fully, two-electron reduced EES to ferric iron oxide. We show how free energy relationships aid in identifying controls on microbial iron oxide reduction by EES, thereby advancing a more fundamental understanding of anaerobic respiration using iron oxides.

This study presents a nation-wide assessment of the influence of chemical and pharmaceutical manufacturing (CPM) wastewaters on synthetic organic contaminant (SOC) emissions to Swiss surface waters. Geographic Information System (GIS) based analysis of the presence of CPM in wastewater treatment plant (WWTP) catchments revealed wide distribution of this industrial sector across Switzerland, suggesting that one-third of the 718 Swiss WWTPs may be influenced by CPM wastewaters. To reflect the diversity of this type of wastewaters, we investigated the effluents of 11 WWTPs of diverse sizes and technologies, which treated 0-100% wastewater from a variety of CPM activities. In an extensive sampling campaign, we collected temporally high resolved (i.e., daily) samples for 2-3 months to capture the dynamics of CPM discharges. The > 850 samples were then measured with liquid chromatography high-resolution mass spectrometry (LC-HRMS). Non-target characterization of the LC-HRMS time series datasets revealed that CPM wastewaters left a highly variable and site-specific signature in the effluents of the WWTPs. Particularly, compared to WWTPs with purely domestic input, a larger variety of substances (up to 15 times more compounds) with higher maximum concentrations (1-2 orders of magnitude) and more uncommon substances were found in CPM-influenced effluents. Moreover, in the latter, highly fluctuating discharges often contributed to a substantial fraction of the overall emissions. The largely varying characteristics of CPM discharges between different facilities were primarily related to the type of activities at the industries (i.e., production versus processing of chemicals) as well as to the pre-treatment and storage of CPM wastewaters. Eventually, for one WWTP, LC-HRMS time series were correlated with ecotoxicity time series obtained from bioassays and major toxic components could be identified. Overall, in view of their potential relevance to water quality, a strong focus on SOC discharges from CPM is essential, including the design of situation-specific monitoring, as well as risk assessment and mitigation strategies that consider the variability of industrial emissions.

Activated sludge treatment leverages the ability of microbes to uptake and (co)-metabolize chemicals and has shown promise in eliminating trace organic contaminants (TrOCs) during wastewater treatment. However, targeted interventions to optimize the process are limited as the fundamental drivers of the observed reactions remain elusive. In this work, we present a comprehensive workflow for the identification and characterization of key enzymes involved in TrOCs biotransformation pathways in complex microbial communities. To demonstrate the applicability of the workflow, we investigated the role of the enzymatic group of multicopper oxidases (MCOs) as one putatively relevant driver of TrOCs biotransformation. To this end, we analyzed activated sludge metatranscriptomic data and selected, synthesized, and heterologously expressed three phylogenetically distinct MCO-encoding genes expressed in communities with different TrOCs oxidation potentials. Following the purification of the encoded enzymes, we screened their activities against different substrates. We saw that MCOs exhibit significant activities against selected TrOCs in the presence of the mediator compound 2,2'-Azino-bis-3-ethylbenzthiazoline-6-sulfonic acid and, in some cases, also in the presence of the wastewater contaminant 4'-hydroxy-benzotriazole. In the first case, we identified oxidation products previously reported from activated sludge communities and concluded that in the presence of appropriate mediators, bacterial MCOs could contribute to the biological removal of TrOCs. Similar investigations of other key enzyme systems may significantly advance our understanding of TrOCs biodegradation and assist the rational design of biology-based water treatment strategies in the future.

Oxygenations of aromatic soil and water contaminants with molecular O₂ catalyzed by Rieske dioxygenases are frequent initial steps of biodegradation in natural and engineered environments. Many of these non-heme ferrous iron enzymes are known to be involved in contaminant metabolism, but the understanding of enzyme-substrate interactions that lead to successful biodegradation is still elusive. Here, we studied the mechanisms of O₂ activation and substrate hydroxylation of two nitroarene dioxygenases to evaluate enzyme- and substrate-specific factors that determine the efficiency of oxygenated product formation. Experiments in enzyme assays of 2-nitrotoluene dioxygenase (2NTDO) and nitrobenzene dioxygenase (NBDO) with methyl-, fluoro-, chloro-, and hydroxy-substituted nitroaromatic substrates reveal that typically 20-100% of the enzyme's activity involves unproductive paths of O₂ activation with generation of reactive oxygen species through so-called O₂ uncoupling. The ¹⁸O and ¹³C kinetic isotope effects of O₂ activation and nitroaromatic substrate hydroxylation, respectively, suggest that O₂ uncoupling occurs after generation of Fe^III-(hydro)peroxo species in the catalytic cycle. While 2NTDO hydroxylates ortho-substituted nitroaromatic substrates more efficiently, NBDO favors meta-substituted, presumably due to distinct active site residues of the two enzymes. Our data implies, however, that the O₂ uncoupling and hydroxylation activity cannot be assessed from simple structure-reactivity relationships. By quantifying O₂ uncoupling by Rieske dioxygenases, our work provides a mechanistic link between contaminant biodegradation, the generation of reactive oxygen species, and possible adaptation strategies of microorganisms to the exposure of new contaminants.

Monitoring changes in stable oxygen isotope ratios in molecular oxygen allows for studying many fundamental processes in bio(geo)chemistry and environmental sciences. While the measurement of ¹⁸O/¹⁶O ratios of O₂ in gaseous samples can be carried out conveniently and from extracting moderately small aqueous samples for analyses by continuous-flow isotope ratio mass spectrometry (CF-IRMS), oxygen isotope signatures, δ¹⁸O, could be overestimated by more than 6‰ because of interferences from argon in air. Here, we systematically evaluated the extent of such Ar interferences on ¹⁸O/¹⁶O ratios of O₂ for measurements by gas chromatography/IRMS and GasBench/IRMS and propose simple instrumental modifications for improved Ar and O₂ separation as well as post-measurement correction procedures for obtaining accurate δ¹⁸O. We subsequently evaluated the consequences of Ar interferences for the quantification of O isotope fractionation in terms of isotope enrichment factors, ϵ_O, and ¹⁸O kinetic isotope effects (¹⁸O KIEs) in samples where O₂ is consumed and Ar:O₂ ratios increase steadily and substantially over the course of a reaction. We show that the extent of O isotope fractionation is overestimated only slightly and that this effect is typically smaller than uncertainties originating from the precision of δ¹⁸O measurements and experimental variability. Ar interferences can become more relevant and bias ϵ_O values by more than 2‰ in aqueous samples where fractional O₂ conversion exceeds 90%. Practically, however, such samples would typically contain less than 25 μM of O₂ at ambient temperature, an amount that is close to the method detection limit of ¹⁸O/¹⁶O ratio measurement by CF-IRMS.

The chemical pollution crisis severely threatens human and environmental health globally. To tackle this challenge the establishment of an overarching international science–policy body has recently been suggested. We strongly support this initiative based on the awareness that humanity has already likely left the safe operating space within planetary boundaries for novel entities including chemical pollution. Immediate action is essential and needs to be informed by sound scientific knowledge and data compiled and critically evaluated by an overarching science–policy interface body. Major challenges for such a body are (i) to foster global knowledge production on exposure, impacts and governance going beyond data-rich regions (e.g., Europe and North America), (ii) to cover the entirety of hazardous chemicals, mixtures and wastes, (iii) to follow a one-health perspective considering the risks posed by chemicals and waste on ecosystem and human health, and (iv) to strive for solution-oriented assessments based on systems thinking. Based on multiple evidence on urgent action on a global scale, we call scientists and practitioners to mobilize their scientific networks and to intensify science–policy interaction with national governments to support the negotiations on the establishment of an intergovernmental body based on scientific knowledge explaining the anticipated benefit for human and environmental health.

1. Freshwater is vital to much life on Earth and is an essential resource for humans. Climate change, however, dramatically changes freshwater systems and reduces water quality, poses a risk to drinking water availability and has severe impacts on aquatic ecosystems and their biodiversity.
2. The direct effects of climate change, such as increased temperatures and higher frequency of extreme meteorological events, interact with human responses to climate change, which we refer to here as 'indirect effects'. The latter possibly have even greater impact than the direct effects of climate change. Specifically, changes in land-use practices as responses to climate change, such as adjusted cropping regimes or a shift to renewable hydroelectricity to mitigate climate change, can very strongly affect freshwater ecosystems.
3. Hitherto, these indirect effects and the possibility of idiosyncratic outcomes are under-recognized. Here, we synthesize knowledge and identify threats to freshwater environments in alpine and pre-alpine regions, which are particularly affected by climate change.
4. We focus on the effects of adapted agriculture and hydropower production on freshwater quality and ecological status, as these examples have strong indirect effects that interact with direct effects of climate change (e.g., water temperature, droughts, isolation of populations).
5. We outline how failure to effectively account for indirect effects associated with human responses to climate change may exacerbate direct climate change impacts on aquatic ecosystems. If managed properly, however, human responses to indirect effects offer potential for rapid and implementable leverage to mitigate some of the direct climate change effects on aquatic ecosystems. To better address looming risks, policy- and decisionmakers must account for indirect effects and incorporate them into restoration planning and the respective sectorial policies.

Auf den Kläranlagen Furt/Bülach und Glarnerland des Abwasserverbandes Glarnerland (AVG, Bilten) wurden zwischen 2014 und 2019 umfangreiche Untersuchungen zur Elimination von Mikroverunreinigungen mittels granulierter Aktivkohle (GAK) durchgeführt. Ziel war es, unter Schweizer Praxisbedingungen die Machbarkeit, Effizienz und Wirtschaftlichkeit des Verfahrens zu evaluieren. Fazit beider Forschungsprojekte ist, dass sich die GAK-Filtration zur Spurenstoffelimination gemäss Schweizer Gesetzgebung eignet. Das Verfahren ist technisch und wirtschaftlich realisierbar und hat sich als praxistauglich erwiesen.

Effluents of wastewater treatment plants can impact microbial communities in the receiving streams. However, little is known about the role of microorganisms in wastewater as opposed to other wastewater constituents, such as nutrients and micropollutants. We aimed therefore at determining the impact of wastewater microorganisms on the microbial diversity and function of periphyton, key microbial communities in streams. We used a flow-through channel system to grow periphyton upon exposure to a mixture of stream water and unfiltered or ultra-filtered wastewater. Impacts were assessed on periphyton biomass, activities and tolerance to micropollutants, as well as on microbial diversity. Our results showed that wastewater microorganisms colonized periphyton and modified its community composition, resulting for instance in an increased abundance of Chloroflexi and a decreased abundance of diatoms and green algae. This led to shifts towards heterotrophy, as suggested by the changes in nutrient stoichiometry and the increased mineralization potential of carbon substrates. An increased tolerance towards micropollutants was only found for periphyton exposed to unfiltered wastewater but not to ultra-filtered wastewater, suggesting that wastewater microorganisms were responsible for this increased tolerance. Overall, our results highlight the need to consider the role of wastewater microorganisms when studying potential impacts of wastewater on the receiving water body.

Rieske dioxygenases belong to the non-heme iron family of oxygenases and catalyze important cis-dihydroxylation as well as O-/N-dealkylation and oxidative cyclization reactions for a wide range of substrates. The lack of substrate coordination at the non-heme ferrous iron center, however, makes it particularly challenging to delineate the role of the substrate for productive O₂ activation. Here, we studied the role of the substrate in the key elementary reaction leading to O₂ activation from a theoretical perspective by systematically considering (i) the 6-coordinate to 5-coordinate conversion of the non-heme Fe^II upon abstraction of a water ligand, (ii) binding of O₂, and (iii) transfer of an electron from the Rieske cluster. We systematically evaluated the spin-state-dependent reaction energies and structural effects at the active site for all combinations of the three elementary processes in the presence and absence of substrate using naphthalene dioxygenase as a prototypical Rieske dioxygenase. We find that reaction energies for the generation of a coordination vacancy at the non-heme Fe^II center through thermoneutral H₂O reorientation and exothermic O₂ binding prior to Rieske cluster oxidation are largely insensitive to the presence of naphthalene and do not lead to formation of any of the known reactive Fe-oxygen species. By contrast, the role of the substrate becomes evident after Rieske cluster oxidation and exclusively for the 6-coordinate non-heme Fe^II sites in that the additional electron is found at the substrate instead of at the iron and oxygen atoms. Our results imply an allosteric control of the substrate on Rieske dioxygenase reactivity to happen prior to changes at the non-heme Fe in agreement with a strategy that avoids unproductive O₂ activation.

Les résultats des mesures NAWA MP 2019 relatives aux insecticides pyréthrinoïdes et organophophorés montrent qu'ils sont détectés dans tous les cours d’eau surveillés à des concentrations certes faibles mais qui représentent des risques importants pour les organismes aquatiques. En effet, les critères de qualité écotoxicologiques ont été dépassés 248 fois de mars à octobre 2019 dans les 17 stations considérées. Ces résultats confirment que sept substances actives d’insecticides représentaient plus de 60% des risques totaux liés à environ 60 pesticides.

Die Ergebnisse des Monitorings von Pyrethroid- und Organophosphat-Insektiziden (Pyr & OrgP) an 17 NAWA-MV-Messstellen im Jahr 2019 erlauben eine erste schweizweite Einschätzung der Gewässerbelastung durch diese Insektizide. Sie zeigen, dass die Insektizide Pyr & OrgP häufig und in allen untersuchten Gewässern gefunden werden, wobei mindestens 3 Substanzen pro Messstelle nachgewiesen wurden. Obwohl die gemessenen Konzentrationen überwiegend unterhalb von 1 ng/l liegen, stellen sie ein erhebliches Risiko für Gewässerorganismen dar. Die ökotoxikologischen Qualitätskriterien wurden insgesamt 248-mal überschritten. Die Ergebnisse zeigen auch, dass sieben Insektizide mehr als 60% des Gesamtrisikos von etwa 60 Pestiziden ausmachten.
Die hier vorgestellten Ergebnisse aus dem Jahr 2019 zeigen zum ersten Mal einen schweizweiten Überblick über die Pyr & OrgPExposition. Änderungen bei der Zulassung von Insektiziden und neue ökotoxikologische Erkenntnisse können zu einer erheblichen Änderung bei der Beurteilung des Gesamtrisikos dieser Insektizide führen.

The evaluation of a chemical substance's persistence is key to understanding its environmental fate, exposure concentration, and, ultimately, environmental risk. Traditional biodegradation test methods were developed many years ago for soluble, nonvolatile, single-constituent test substances, which do not represent the wide range of manufactured chemical substances. In addition, the Organisation for Economic Co-operation and Development (OECD) screening and simulation test methods do not fully reflect the environmental conditions into which substances are released and, therefore, estimates of chemical degradation half-lives can be very uncertain and may misrepresent real environmental processes. In this paper, we address the challenges and limitations facing current test methods and the scientific advances that are helping to both understand and provide solutions to them. Some of these advancements include the following: (1) robust methods that provide a deeper understanding of microbial composition, diversity, and abundance to ensure consistency and/or interpret variability between tests; (2) benchmarking tools and reference substances that aid in persistence evaluations through comparison against substances with well-quantified degradation profiles; (3) analytical methods that allow quantification for parent and metabolites at environmentally relevant concentrations, and inform on test substance bioavailability, biochemical pathways, rates of primary versus overall degradation, and rates of metabolite formation and decay; (4) modeling tools that predict the likelihood of microbial biotransformation, as well as biochemical pathways; and (5) modeling approaches that allow for derivation of more generally applicable biotransformation rate constants, by accounting for physical and/or chemical processes and test system design when evaluating test data. We also identify that, while such advancements could improve the certainty and accuracy of persistence assessments, the mechanisms and processes by which they are translated into regulatory practice and development of new OECD test guidelines need improving and accelerating. Where uncertainty remains, holistic weight of evidence approaches may be required to accurately assess the persistence of chemicals.

Biotransformation is the most important process removing manmade chemicals from the environment, yet mechanisms governing this essential ecosystem function are underexplored. To understand these mechanisms, we conducted experiments in flow-through systems, by colonizing stream biofilms under different conditions of mixing river water with treated (and ultrafiltered) wastewater. We performed biotransformation experiments with those biofilms, using a set of 75 micropollutants, and could disentangle potential mechanisms determining the biotransformation potential of stream biofilms. We showed that the increased biotransformation potential downstream of wastewater treatment plants that we observed for specific micropollutants contained in household wastewaters (downstream effect) is caused by microorganisms released with the treated effluent, rather than by the in-stream exposure to those micropollutants. Complementary data from 16S rRNA amplicon-sequencing revealed 146 amplicon sequence variants (ASVs) that followed the observed biotransformation patterns. Our results align with findings for community tolerance, and provide clear experimental evidence that microorganisms released with treated wastewater integrate into downstream biofilms and impact crucial ecosystem functions.

Das diesjährige Bundeslager der Pfadibewegung Schweiz fand im Goms statt. Die über 30 000 Teilnehmenden wurde über ein eigens dafür gebautes temporäres Netz mit Trinkwasser versorgt. Mit einem umfassenden Selbstkontrollkonzept wurde ein qualitativ einwandfreies Trinkwasser sichergestellt. Zentrale Punkte waren dabei eine Gefahren- und Risikoanalyse, Kontrollen auf Platz mittels Online- und Feldmessungen sowie Laboranalysen.

Bioaccumulation and trophic transfer of persistent legacy contaminants have been intensively characterized, but little is known on the contaminants of emerging concern (CECs) in freshwater food webs. Herein, we comprehensively screened CECs with a focus on polar substances and further evaluated their trophic transfer behavior in selected items from the food web of Lake Templin, Germany. Weselected one plankton, two mussel, and nine fish samples covering three trophic levels. With an effective multi-residue sample preparation method and high-resolution mass spectrometry-based target, suspect, and non-target screening, we characterized 477 targets and further screened unknown features in complex biota matrices. Of the 477 targets, 145 were detected and quantified in at least one species (0.02–3640 ng/g, dry weight). Additionally, the suspect and non-target analysis with experimental mass spectra libraries and in silico techniques (MetFrag and SIRIUS4/CSI:FingerID) enabled further identification of 27 unknown compounds with 19 confirmed by reference standards. Overall, the detected compounds belong to a diverse group of chemicals, including 71 pharmaceuticals, 27 metabolites, 26 pesticides, 16 per- and polyfluoroalkyl substances (PFASs), 4 plasticizers, 3 flame retardants, 11 other industrial chemicals and 14 others. Moreover, we determined the trophic magnification factor (TMF) of 34 polar CECs with >80% detection frequency, among which 6 PFASs including perfluorooctane sulfonic acid (PFOS), perfluorodecanoic acid (PFDA), perfluorohexane sulfonic acid (PFHxS), perfluorotridecanoic acid (PFTrA), perfluorotetradecanoic acid (PFTeA), and perfluoroundecanoic acid (PFUnA), exhibited biomagnification potential (TMF =1.8 - 4.2, p < 0.05), whereas 5 pharmaceuticals (phenazone, progesterone, venlafaxine, levamisole, and lidocaine) and 1 personal care product metabolite (galaxolidone) showed biodilution potential (TMF = 0.4 - 0.6, p < 0.05).

Background: Poorly educated smallholder farmers in low-income countries are highly exposed to pesticides. This can result in adverse mental health issues, of which sleep problems might be an underlying indicator. We aim to examine the association between sleep problems and pesticide exposure among smallholder farmers in Uganda.
Methods: A cross-sectional survey with 253 smallholder farmers was conducted between October and December 2019. Sleep problems were assessed during the week before the visit using the Medical Outcomes Study Sleep Scale (MOS-SS). Exposure to pesticides was assessed as application days of any pesticide and as use of 2,4-D, glyphosate, mancozeb, organophosphates & carbamates, pyrethroids and other pesticides during the week and year prior to the visit. Associations were assessed using adjusted multivariable logistic regression models.
Results: Increased odds ratio (OR) for the sleep problem index 6-items (OR [95% Confidence Interval] 1.99 [1.04; 3.84] and 3.21 [1.33; 7.82]), sleep inadequacy (1.94 [1.04; 3.66] and 2.49 [1.05-6.22]) and snoring (3.17 [1.12; 9.41] and 4.07 [1.04; 15.14]) were observed for farmers who respectively applied pesticides up to two days and three or more days in the past week compared to farmers who did not apply during the past week. Gender-stratified analyses showed a higher OR for female applicators (4.27 [1.76-11.16]) than for male applicators (1.82 [0.91–3.79]) for the association between the sleep problem index 6-items and pesticide use in the week before the visit. Increased ORs were also observed for the association between the sleep problem index 6-item and mancozeb exposure during the past year 2.28 [1.12-4.71] and past week 2.51 [0.86-7.55] and glyphosate exposure during the past week 3.75 [1.24-11.8] compared to non-applicators.
Discussion: Our findings suggest an increased risk of sleep problems among smallholder farmers in a pesticide-exposure-dependent way in a low-income context. Further gender-stratified, longitudinal investigations are warranted to confirm these findings.

Over the past three decades, the Vietnamese Mekong Delta has experienced a significant increase in agricultural productivity, partly achieved through increased agrochemical use. To abate negative effects on human and environmental health, several national programs were launched to enhance safer pesticide use. This study aimed to assess the patterns and relationships of official sustainable agriculture educational programs, pesticide safety knowledge, and practices of smallholder farmers in the Mekong Delta. A cross-sectional survey was conducted with 400 smallholder farmers from three communes in Thoi Lai district (Can Tho province) from March to May 2020. Twenty-four questions on pesticide safety knowledge and practices were used to identify traits using latent class analysis. Adjusted generalized linear regression was used to assess determinants of pesticide safety knowledge and estimate associations of pesticide safety knowledge with pesticide practices. 96.2% of participants have used at least one WHO class II pesticide during the past year while the use of specific personal protective equipment was limited mainly due to unavailability (37.0%) or discomfort (83.0%). High education (Odds Ratio (OR), 95% Confidence Interval; 3.84, 1.70-9.45), exposure to official educational programs (1.87, 1.13-3.12), peer-to-peer knowledge exchange (3.58, 2.18-6.00), and learning from governmental extension services (2.31, 1.14-4.98) were positively associated with increased pesticide safety knowledge. Compared to poor practices, pesticide safety knowledge was increasingly positively associated with intermediate (1.65, 1.02-2.66) and good pesticide practices (8.96, 2.58-31.12). These findings highlight the importance of school education and educational programs, access to PPE, and addressing discomforts of PPE to improve the protection of farmers from pesticide exposures. Simultaneously, pesticide market authorization processes should be reconsidered to promote the authorization of less toxic products. Further in-depth studies on the nature of pesticides used, nonuse of personal protective equipment, and effectiveness of educational programs will further define leverage points for safer pesticide use.

Oxidative treatment of seawater in coastal and shipboard installations is applied to control biofouling and/or minimize the input of noxious or invasive species into the marine environment. This treatment allows a safe and efficient operation of industrial installations and helps to protect human health from infectious diseases and to maintain the biodiversity in the marine environment. On the downside, the application of chemical oxidants generates undesired organic compounds, so-called disinfection by-products (DBPs), which are discharged into the marine environment. This article provides an overview on sources and quantities of DBP inputs, which could serve as basis for hazard analysis for the marine environment, human health and the atmosphere. During oxidation of marine water, mainly brominated DBPs are generated with bromoform (CHBr₃) being the major DBP. CHBr₃ has been used as an indicator to compare inputs from different sources. Total global annual volumes of treated seawater inputs resulting from cooling processes of coastal power stations, from desalination plants and from ballast water treatment in ships are estimated to be 470-800 × 10⁹ m³, 46 × 10⁹ m³ and 3.5 × 10⁹ m³, respectively. Overall, the total estimated anthropogenic bromoform production and discharge adds up to 13.5-21.8 × 10⁶ kg/a (kg per year) with contributions of 11.8-20.1 × 10⁶ kg/a from cooling water treatment, 0.89 × 10⁶ kg/a from desalination and 0.86 × 10⁶ kg/a from ballast water treatment. This equals approximately 2–6% of the natural bromoform emissions from marine water, which is estimated to be 385-870 × 10⁶ kg/a.

The Madeira is one of the major tributaries of the Amazon River and is characterized by a large alluvial floodplain throughout the stream continuum. This study aims to better assess the hydrological functioning of the Madeira Basin over its alluvial floodplains at both local and global scales. We used the semi-distributed hydrological Soil and Water Assessment Tool (SWAT) model to simulate water discharge at a daily time step and water resources for each hydrological compartment. A new hydraulic module for water routing was implemented in the SWAT model considering the floodplain either as a simple reservoir or as a continuum where the water can flow along with the floodplain network. Both water surface estimated by L-band passive microwaves (SWAF data) and digital elevation model—shuttle radar topography mission (DEM–SRTM data) were used to delineate the floodplain, as inputs for the model. On the global scale, the amount of water stored in the Madeira floodplain is between 810 ± 230 km³ per year when the floodplains are delimited with SWAF and 1,300 ± 350 km³ per year with the DEM floodplain delineation between 2008 and 2018. Spatial altimetry (Jason 2-3) data were also applied to alluvial areas to validate the water height dynamic in floodplains at a local scale. Results show that more than 60% of the alluvial validation points display a correlation above 0.40 ± 0.02 regardless of the floodplain delineation. This study permits us to better characterize the spatio-temporal storage dynamics of the Madeira floodplains at both local and global scales, and it underlines the importance of a precise floodplain delineation, before computing biogeochemical fluxes and sediment yield.

Background: Safe and clean drinking water is essential for human life. Persistent, mobile and toxic (PMT) substances and/or very persistent and very mobile (vPvM) substances are an important group of substances for which additional measures to protect water resources may be needed to avoid negative environmental and human health effects. PMT/vPvM substances do not sufficiently biodegrade in the environment, they can travel long distances with water and are toxic (those that are PMT substances) to the environment and/or human health. PMT/vPvM substance research and regulation is arguably in its infancy and in order to get in control of these substances the following (non-exhaustive list of) knowledge gaps should to be addressed: environmental occurrence; the suitability of currently available analytical methods; the effectiveness and availability of treatment technologies; the ability of regional governance and industrial stewardship to contribute to safe drinking water while supporting innovation; the ways in which policies and regulations can be used most effectively to govern these substances; and, the identification of safe and sustainable alternatives. Methods: The work is the outcome of the third PMT workshop, held in March 2021, that brought together diverse scientists, regulators, NGOs, and representatives from the water sector and the chemical sector, all concerned with protecting the quality of our water resources. The online workshop was attended by over 700 people. The knowledge gaps above were discussed in the presentations given and the attendees were invited to provide their opinions about knowledge gaps related to PMT/vPvM substance research and regulation. Results: Strategies to closing the knowledge, technical and practical gaps to get in control of PMT/vPvM substances can be rooted in the Chemicals Strategy for Sustainability Towards a Toxic Free Environment from the European Commission, as well as recent advances in the research and industrial stewardship. Key to closing these gaps are: (i) advancing remediation and removal strategies for PMT/vPvM substances that are already in the environment, however this is not an effective long-term strategy; (ii) clear and harmonized definitions of PMT/vPvM substances across diverse European and international legislations; (iii) ensuring wider availability of analytical methods and reference standards; (iv) addressing data gaps related to persistence, mobility and toxicity of chemical substances, particularly transformation products and those within complex substance mixtures; and (v) advancing monitoring and risk assessment tools for stewardship and regulatory compliance. The two most effective ways to get in control were identified to be source control through risk governance efforts, and enhancing market incentives for alternatives to PMT/vPvM substances by using safe and sustainable by design strategies.

The frequency and intensity of cyanobacterial blooms continue to increase in freshwater systems across the globe. Cyanobacteria can release toxins and several bioactive secondary metabolites and analytical methods are needed to effectively assess their concentrations in surface waters. Since booms can evolve rapidly in parts of a lake, high resolution of spatial and temporal sampling increases the complexity of monitoring efforts. Here, we present the validation of an automated, online-solid phase extraction (SPE) high performance liquid chromatography (HPLC)-high resolution tandem mass spectrometry (HRMS/MS) method. This online-SPE HPLC-HRMS/MS methods enables quantitative monitoring of surface waters for 17 cyanobacterial peptides (cyanopeptides), spanning 5 distinct cyanopeptide classes, including: microcystins, anabaenopeptins, nodularins, cyclamides and cyanopeptolins. The method can quantify these cyanopeptides in the low ng/L-range with high accuracy (85-116%) and low relative matrix effects (<25%). We demonstrated its application to Swiss lake waters (Zürichsee, Hallwilersee, Greifensee), which also highlighted the value of adding cyanopeptides beyond common microcystins when monitoring surface waters for cyanobacteria.

Monitoring biodiversity is essential to understand the impacts of human activities and for effective management of ecosystems. Thereby, biodiversity can be assessed through direct collection of targeted organisms, through indirect evidence of their presence (e.g. signs, environmental DNA, camera trap, etc.), or through extrapolations from species distribution and species richness models. Differences in approaches used in biodiversity assessment, however, may come with individual challenges and hinder cross-study comparability. In the context of rapidly developing techniques, we compared three different approaches in order to better understand assessments of aquatic macroinvertebrate diversity. Specifically, we compared the community composition and species richness of three orders of aquatic macroinvertebrates (mayflies, stoneflies, and caddisflies, hereafter EPT) obtained via eDNA metabarcoding and via traditional in situ kicknet sampling to catchment-level based predictions of a species richness model. We used kicknet data from 24 sites in Switzerland and compared taxonomic lists to those obtained using eDNA amplified with two different primer sets. Richness detected by these methods was compared to the independent predictions made by a statistical species richness model, that is, a generalized linear model using landscape-level features to estimate EPT diversity. Despite the ability of eDNA to consistently detect some EPT species found by traditional sampling, we found important discrepancies in community composition between the kicknet and eDNA approaches, particularly at a local scale. We found the EPT-specific primer set fwhF2/EPTDr2n, detected a greater number of targeted EPT species compared to the more general primer set mlCOIintF/HCO2198. Moreover, we found that the species richness measured by eDNA from either primer set was poorly correlated to the richness measured by kicknet sampling (Pearson correlation = 0.27) and that the richness estimated by eDNA and kicknet were poorly correlated with the prediction of the species richness model (Pearson correlation = 0.30 and 0.44, respectively). The weak relationships between the traditional kicknet sampling and eDNA with this model indicates inherent limitations in upscaling species richness estimates, and possibly a limited ability of the model to meet real world expectations. It is also possible that the number of replicates was not sufficient to detect ambiguous correlations. Future challenges include improving the accuracy and sensitivity of each approach individually, yet also acknowledging their respective limitations, in order to best meet stakeholder demands and address the biodiversity crisis we are facing.

Micropollutants present in the effluent of wastewater treatment plants (WWTPs) after biological treatment are largely eliminated by effective advanced technologies such as ozonation. Discharge of contaminants into freshwater ecosystems can thus be minimized, while simultaneously protecting drinking water resources. However, ozonation can lead to reactive and potentially toxic transformation products. To remove these, the Swiss Federal Office for the Environment recommends additional "post-treatment" of ozonated WWTP effluent using sand filtration, but other treatments may be similarly effective. In this study, 48 h composite wastewater samples were collected before and after full-scale ozonation, and after post-treatments (full-scale sand filtration, pilot-scale fresh and pre-loaded granular activated carbon, and fixed and moving beds). Ecotoxicological tests were performed to quantify the changes in water quality following different treatment steps. These included standard in vitro bioassays for the detection of endocrine, genotoxic and mutagenic effects, as well as toxicity to green algae and bacteria, and flow-through in vivo bioassays using oligochaetes and early life stages of rainbow trout.
Results show that ozonation reduced a number of ecotoxicological effects of biologically treated wastewater by 66 - 93%: It improved growth and photosynthesis of green algae, decreased toxicity to luminescent bacteria, reduced concentrations of hormonally active contaminants and significantly changed expression of biomarker genes in rainbow trout liver. Bioassay results showed that ozonation did not produce problematic levels of reaction products overall. Small increases in toxicity observed in a few samples were reduced or eliminated by post-treatments. However, only relatively fresh granular activated carbon (analyzed at 13,000 - 20,000 bed volumes) significantly reduced effects additionally (by up to 66%) compared to ozonation alone. Inhibition of algal photosynthesis, rainbow trout liver histopathology and biomarker gene expression proved to be sufficiently sensitive endpoints to detect the change in water quality achieved by post-treatment.

Photo-induced cross-linking is a mainstay technique to characterize RNA-protein interactions. However, UV-induced cross-linking between RNA and proteins at “zero-distance” is poorly understood. Here, we investigate cross-linking of the RBFOX alternative splicing factor with its hepta-ribonucleotide binding element as a model system. We examine the influence of nucleobase, nucleotide position and amino acid composition using CLIR-MS technology (crosslinking-of-isotope-labelled-RNA-and-tandem-mass-spectrometry), that locates cross-links on RNA and protein with site-specific resolution. Surprisingly, cross-linking occurs only at nucleotides that are π-stacked to phenylalanines. Notably, this π-stacking interaction is also necessary for the amino-acids flanking phenylalanines to partake in UV-cross-linking. We confirmed these observations in several published datasets where cross-linking sites could be mapped to a high resolution structure. We hypothesize that π-stacking to aromatic amino acids activates cross-linking in RNA-protein complexes, whereafter nucleotide and peptide radicals recombine. These findings will facilitate interpretation of cross-linking data from structural studies and from genome-wide datasets generated using CLIP (cross-linking-and-immunoprecipitation) methods.

Pesticides used in agriculture can end up in nearby streams and can have a negative impact on nontarget organisms such as aquatic invertebrates. During registration, bioaccumulation potential is often investigated using laboratory tests only. Recent studies showed that the magnitude of bioaccumulation in the field substantially differs from laboratory conditions. To investigate this discrepancy, we conducted a field bioaccumulation study in a stream known to receive pollutant loadings from agriculture. Our work incorporates measurements of stream pesticide concentrations at high temporal resolution (every 20 min), as well as sediment, leaves, and caged gammarid analyses (every 2-24 h) over several weeks. Of 49 investigated pesticides, 14 were detected in gammarids with highly variable concentrations of up to 140 ± 28 ng/g_ww. Toxicokinetic modeling using laboratory-derived uptake and depuration rate constants for azoxystrobin, cyprodinil, and fluopyram showed that despite the highly resolved water concentrations measured, the pesticide burden on gammarids remains underestimated by a factor of 1.9 ± 0.1 to 31 ± 3.0, with the highest underestimations occurring after rain events. Including dietary uptake from polluted detritus leaves and sediment in the model explained this underestimation only to a minor proportion. However, suspended solids analyzed during rain events had high pesticide concentrations, and uptake from them could partially explain the underestimation after rain events. Additional comparison between the measured and modeled data showed that the pesticide depuration in gammarids is slower in the field. This observation suggests that several unknown mechanisms may play a role, including lowered enzyme expression and mixture effects. Thus, it is important to conduct such retrospective risk assessments based on field investigations and adapt the registration accordingly.

Leverett et al. commented on the Environmental quality standard (EQS) for diclofenac derived under the European Water Framework Directive [Leverett et al. (2021) Environ Sci Eur 33: 133 https://doi.org/10.1186/s12302-021-00574-z]. They postulated that the derivation of the EQS value for diclofenac is not conducted according to the EQS Technical Guidance, but rather using data of poor reliability and relevance. Consequently, the authors suggested using their alternative derived value instead. It is to be noted that the process for the EQS derivation for diclofenac is still ongoing and not finalized, and that as a consequence, any critical analysis is very premature. In general, within the current European Commission process, EQS values proposals are derived by expert groups led by the Joint Research Centre. In the specific case for diclofenac, Leverett et al. have also been actively involved as experts. This response to Leverett et al. (2021) aims to clarify the reasoning behind the proposal from a scientific point of view and to express our concern for the lack of transparency of their position in the statement of competing interests. Indeed, the authors did not disclose their participation in the expert group for deriving the diclofenac EQS value, nor that they have direct and indirect ties to a company that markets diclofenac in Europe, Glaxo Smith & Kline plc (GSK). This amounts to a significant conflict of interest and leads to disinformation to the reader.

Metabolic "dark matter" describes currently unknown metabolic processes, which form a blind spot in our general understanding of metabolism and slow down the development of biosynthetic cell factories and naturally derived pharmaceuticals. Mapping the dark matter of metabolism remains an open challenge that can be addressed globally and systematically by existing computational solutions. In this work, we use 489 generalized enzymatic reaction rules to map both known and unknown metabolic processes around a biochemical database of 1.5 million biological compounds. We predict over 5 million reactions and integrate nearly 2 million naturally and synthetically-derived compounds into the global network of biochemical knowledge, named ATLASx. ATLASx is available to researchers as a powerful online platform that supports the prediction and analysis of biochemical pathways and evaluates the biochemical vicinity of molecule classes (https://lcsb-databases.epfl.ch/Atlas2).

Background: Previous epidemiological studies have reported associations of pesticide exposure with poor cognitive function and behavioral problems. However, these findings have relied primarily on neuropsychological assessments. Questions remain about the neurobiological effects of pesticide exposure, specifically where in the brain pesticides exert their effects and whether compensatory mechanisms in the brain may have masked pesticide-related associations in studies that relied purely on neuropsychological measures.
Methods: We conducted a functional neuroimaging study in 48 farmworkers from Zarcero County, Costa Rica, in 2016. We measured concentrations of 13 insecticide, fungicide, or herbicide metabolites or parent compounds in urine samples collected during two study visits (approximately 3-5 weeks apart). We assessed cortical brain activation in the prefrontal cortex during tasks of working memory, attention, and cognitive flexibility using functional near-infrared spectroscopy (fNIRS). We estimated associations of pesticide exposure with cortical brain activation using multivariable linear regression models adjusted for age and education level.
Results: We found that higher concentrations of insecticide metabolites were associated with reduced activation in the prefrontal cortex during a working memory task. For example, 3,5,6-trichloro-2-pyridinol (TCPy; a metabolite of the organophosphate chlorpyrifos) was associated with reduced activation in the left dorsolateral prefrontal cortex (β = -2.3; 95% CI: -3.9, -0.7 per two-fold increase in TCPy). Similarly, 3-phenoxybenzoic acid (3-PBA; a metabolite of pyrethroid insecticides) was associated with bilateral reduced activation in the dorsolateral prefrontal cortices (β = -3.1; 95% CI: -5.0, -1.2 and -2.3; 95% CI: -4.5, -0.2 per two-fold increase in 3-PBA for left and right cortices, respectively). These associations were similar, though weaker, for the attention and cognitive flexibility tasks. We observed null associations of fungicide and herbicide biomarker concentrations with cortical brain activation during the three tasks that were administered.
Conclusion: Our findings suggest that organophosphate and pyrethroid insecticides may impact cortical brain activation in the prefrontal cortex – neural dynamics that could potentially underlie previously reported associations with cognitive and behavioral function. Furthermore, our study demonstrates the feasibility and utility of fNIRS in epidemiological field studies.

Rieske dioxygenases catalyze the initial steps in the hydroxylation of aromatic compounds and are critical for the metabolism of xenobiotic substances. Because substrates do not bind to the mononuclear non-heme Fe^II center, elementary steps leading to O₂ activation and substrate hydroxylation are difficult to delineate, thus making it challenging to rationalize divergent observations on enzyme mechanisms, reactivity, and substrate specificity. Here, we show for nitrobenzene dioxygenase, a Rieske dioxygenase capable of transforming nitroarenes to nitrite and substituted catechols, that unproductive O₂ activation with the release of the unreacted substrate and reactive oxygen species represents an important path in the catalytic cycle. Through correlation of O₂ uncoupling for a series of substituted nitroaromatic compounds with ¹⁸O and ¹³C kinetic isotope effects of dissolved O₂ and aromatic substrates, respectively, we show that O₂ uncoupling occurs after the rate-limiting formation of Fe^III-(hydro)peroxo species from which substrates are hydroxylated. Substituent effects on the extent of O₂ uncoupling suggest that the positioning of the substrate in the active site rather than the susceptibility of the substrate for attack by electrophilic oxygen species is responsible for unproductive O₂ uncoupling. The proposed catalytic cycle provides a mechanistic basis for assessing the very different efficiencies of substrate hydroxylation vs unproductive O₂ activation and generation of reactive oxygen species in reactions catalyzed by Rieske dioxygenases.

Cyanobacteria, also known as blue-green algae, are photosynthetic bacteria that can colonize different habitats, including extreme ones. They are of great interest to the scientific community, especially because of their ability to produce cyanotoxins: toxic secondary metabolites potentially harmful to organisms especially when released to surface waters.

Liquid chromatography-mass spectrometry (LC-MS)-based untargeted metabolomics experiments have become increasingly popular because of the wide range of metabolites that can be analyzed and the possibility to measure novel compounds. LC-MS instrumentation and analysis conditions can differ substantially among laboratories and experiments, thus resulting in non-standardized datasets demanding customized annotation workflows. We present an ecosystem of R packages, centered around the MetaboCoreUtils, MetaboAnnotation and CompoundDb packages that together provide a modular infrastructure for the annotation of untargeted metabolomics data. Initial annotation can be performed based on MS¹ properties such as m/z and retention times, followed by an MS²-based annotation in which experimental fragment spectra are compared against a reference library. Such reference databases can be created and managed with the CompoundDb package. The ecosystem supports data from a variety of formats, including, but not limited to, MSP, MGF, mzML, mzXML, netCDF as well as MassBank text files and SQL databases. Through its highly customizable functionality, the presented infrastructure allows to build reproducible annotation workflows tailored for and adapted to most untargeted LC-MS-based datasets. All core functionality, which supports base R data types, is exported, also facilitating its re-use in other R packages. Finally, all packages are thoroughly unit-tested and documented and are available on GitHub and through Bioconductor.

Das Monitoring von Pflanzenschutzmittel-Wirkstoffen und -Metaboliten im Grundwasser gehört zu den Kernaufgaben der Nationalen Grundwasserbeobachtung NAQUA. Die Zahl der Substanzen, die im Langzeitmonitoring analysiert werden können, ist allerdings limitiert. Umso wichtiger ist eine gezielte und fundierte Auswahl bzw. Priorisierung der Substanzen.

The intensive use of pesticides and their subsequent distribution to the environment and non-target organisms is of increasing concern. So far, little is known about the occurrence of pesticides in soils of untreated areas- such as ecological refuges- as well as the processes contributing to this unwanted pesticide contamination. In this study, we analyzed the presence and abundance of 46 different pesticides in soils from extensively managed grassland sites, as well as organically and conventionally managed vegetable fields (60 fields in total). Pesticides were found in all soils, including the extensive grassland sites, demonstrating a widespread background contamination of soils with pesticides. The results suggest that after conversion from conventional to organic farming, the organic fields reach pesticide levels as low as those of grassland sites not until 20 years later. Furthermore, the different pesticide composition patterns in grassland sites and organically managed fields facilitated differentiation between long-term persistence of residues and diffuse contamination processes, that is, short-scale redistribution (spray drift) and long-scale dispersion (atmospheric deposition), to offsite contamination.

Cross-linking and mass spectrometry (XL-MS) workflows represent an increasingly popular technique for low-resolution structural studies of macromolecular complexes. Cross-linking reactions take place in the solution state, capturing contact sites between components of a complex that represent the native, functionally relevant structure. Protein-protein XL-MS protocols are widely adopted, providing precise localization of cross-linking sites to single amino acid positions within a pair of cross-linked peptides. In contrast, protein-RNA XL-MS workflows are evolving rapidly and differ in their ability to localize interaction regions within the RNA sequence. Here, we review protein-protein and protein-RNA XL-MS workflows, and discuss their applications in studies of protein-RNA complexes. The examples highlight the complementary value of XL-MS in structural studies of protein-RNA complexes, where more established high-resolution techniques might be unable to produce conclusive data.

Surfactants are high-production-volume chemicals that are among the most abundant organic pollutants in municipal wastewater. In this study, sewage sludge samples of 36 Swiss wastewater treatment plants (WWTPs), serving 32% of the country's population, were analyzed for major surfactant classes by liquid chromatography mass spectrometry (LC-MS). The analyses required a variety of complementary approaches due to different analytical challenges, including matrix effects (which can affect adduct ion formation) and the lack of reference standards. The most abundant contaminants were linear alkylbenzene sulfonates (LAS; weighted mean [WM] concentration of 3700 μg g^-1 dry weight), followed by secondary alkane sulfonates (SAS; 190 μg g^-1). Alcohol polyethoxylates (AEO; 8.3 μg g^-1), nonylphenol polyethoxylates (NPEO; 16 μg g^-1), nonylphenol (NP; 3.1 μg g^-1), nonylphenol ethoxy carboxylates (NPEC; 0.35 μg g^-1) and tert-octylphenol (tert-OP, 1.8 μg g^-1) were present at much lower concentrations. This concentration pattern agrees with the production volumes of the surfactants and their fates in WWTPs. Branched AEO homologues dominated over linear homologues, probably due to higher persistence. Sludge concentrations of LAS, SAS, and NP were positively correlated with the residence time in the anaerobic digester. Derivation of the per capita loads successfully revealed potential industrial/commercial emission sources. Comparison of recent versus historic data showed a decrease in NPEO and NP levels by one or two orders of magnitude since their ban in the 1980s. By contrast, LAS still exhibit similar concentrations compared to 30 years ago.

Spray drift is considered a major pesticide transport pathway to surface waters. Current research and legislation usually only considers direct spray drift. However, also spray drift on roads and subsequent wash-off to surface waters was identified as a possible transport pathway. Hydraulic shortcuts (storm drainage inlets, channel drains, ditches) have been shown to connect roads to surface waters, thus increasing the risk of drift wash-off to surface waters. However, the importance of this pathway has never been assessed on larger scales. To address this knowledge gap, we studied 26 agricultural catchments with a predominance of arable cropping (n = 17) and vineyards (n = 9). In these study sites, we assessed the occurrence of shortcuts by field mapping. Afterwards, we modelled the areas of roads drained to surface waters using a high-resolution digital elevation model (0.5 m resolution) and a multiple flow algorithm. Finally, we modelled drift deposition to drained roads and surface waters using a spatially explicit, georeferenced spray drift model. Our results show that for most sites, the drift to drained roads is much larger than the direct drift to surface waters. In arable land sites, drift to roads exceeds the direct drift by a factor of 4.5 to 18, and in vineyard sites by 35 to 140. In arable land sites, drift to drained roads is rather small (0.0015% to 0.0049% of applied amount) compared to typical total pesticide losses to surface waters. However, substantial drift to drained roads in vineyard sites was found (0.063% to 0.20% of applied amount). Current literature suggests that major fractions of the drift deposited on roads can be washed off during rain events, especially for pesticides with low soil adsorption coefficients. For such pesticides and particularly in vineyards, spray drift wash-off from drained roads is therefore expected to be a major transport pathway to surface waters.

Agricultural pesticides transported to surface waters pose a major risk for aquatic ecosystems. Modelling studies indicate that the inlets of agricultural storm drainage systems can considerably increase the connectivity of surface runoff and pesticides to surface waters. These model results have however not yet been validated with field measurements. In this study, we measured discharge and concentrations of 51 pesticides in four out of 158 storm drainage inlets of a small Swiss agricultural catchment (2.8 km²) and in the receiving stream. For this, we performed an event-triggered sampling during 19 rain events and collected plot-specific pesticide application data. Our results show that agricultural storm drainage inlets strongly influence surface runoff and pesticide transport in the study catchment. The concentrations of single pesticides in inlets amounted up to 62 µg/L. During some rain events, transport through single inlets caused more than 10% of the stream load of certain pesticides. An extrapolation to the entire catchment suggests that during selected events on average 30 to 70% of the load in the stream was transported through inlets. Pesticide applications on fields with surface runoff or spray drift potential to inlets led to increased concentrations in the corresponding inlets. Overall, this study corroborates the relevance of such inlets for pesticide transport by establishing a connectivity between fields and surface waters, and by their potential to deliver substantial pesticide loads to surface waters.

Introduction. Pesticides used in agriculture are transported to surface waters through various pathways and pose a major threat to aquatic ecosystems. Farmers and authorities take various measures to reduce pesticide transport to surface waters and to protect them from harmful effects. However, such actions can only be effective if the underlying processes driving the pesticide risk are understood well enough. Previous research suggests that so-called hydraulic shortcuts may be an important pesticide transport pathway that has been overlooked in the past. The term hydraulic shortcuts refers to inlet or maintenance shafts of agricultural storm drainage systems, but also to roads, farm tracks, channel drains, and ditches. Even though the relevance of hydraulic shortcuts for pesticide transport has been shown in single cases, it is unclear how often these structures occur and how relevant they are in general for pesticide transport compared to other pathways.
Objectives. This thesis aimed on quantifying the relevance of pesticide losses via hydraulic shortcuts to Swiss surface waters. For this, the following four research questions were investigated: 1) How often do hydraulic shortcuts occur in Swiss agricultural areas? 2) What is their relevance for surface runoffrelated pesticide transport? 3) What is their relevance for spray drift-related pesticide transport? 4) What pesticide concentrations and loads are found in hydraulic shortcuts? [...]

Phosphorus (P) is an essential element to all living beings but also a finite resource. P-related problems center around broken P cycles from local to global scales. This paper presents outcomes from the 9th International Phosphorus Workshop (IPW9) held 2019 on how to move towards a sustainable P management. It is based on two sequential discussion rounds with all participants. Important progress was reported regarding the awareness of P as finite mineable resource, technologies to recycle P, and legislation towards a circular P economy. Yet, critical deficits were identified such as how to handle legacy P, how climate change may affect ecosystem P cycling, or working business models to up-scale existing recycling models. Workshop participants argued for more transdisciplinary networks to narrow a perceived science-practice/policy gap. While this gap may be smaller in reality as illustrated with a Swiss example, we formulate recommendations how to bridge this gap more effectively.

In den 50 Jahren ihres Bestehens ermöglichte die Nationale Daueruntersuchung der Fliessgewässer (NADUF), zahlreiche Erfolge von Gewässerschutzmassnahmen in der Schweiz quantitativ zu belegen und Zustandsänderungen aufzuzeigen. Dazu gehören reduzierte Einträge von Phosphat, Schwermetallen oder Stickstoffverbindungen in die Gewässer. In der Zukunft stellen sich beim Monitoring neue Herausforderungen wie die Ermittlung der Einträge von organischen Mikroverunreinigungen oder die durch den Klimawandel induzierten Änderungen im aquatischen Ökosystem.

Current methods for structure elucidation of small molecules rely on finding similarity with spectra of known compounds, but do not predict structures de novo for unknown compound classes. We present MSNovelist, which combines fingerprint prediction with an encoder–decoder neural network to generate structures de novo solely from tandem mass spectrometry (MS²) spectra. In an evaluation with 3,863 MS² spectra from the Global Natural Product Social Molecular Networking site, MSNovelist predicted 25% of structures correctly on first rank, retrieved 45% of structures overall and reproduced 61% of correct database annotations, without having ever seen the structure in the training phase. Similarly, for the CASMI 2016 challenge, MSNovelist correctly predicted 26% and retrieved 57% of structures, recovering 64% of correct database annotations. Finally, we illustrate the application of MSNovelist in a bryophyte MS² dataset, in which de novo structure prediction substantially outscored the best database candidate for seven spectra. MSNovelist is ideally suited to complement library-based annotation in the case of poorly represented analyte classes and novel compounds.

Background: The NORMAN Association (https://www.norman-network.com/) initiated the NORMAN Suspect List Exchange (NORMAN-SLE; https://www.norman-network.com/nds/SLE/) in 2015, following the NORMAN collaborative trial on non-target screening of environmental water samples by mass spectrometry. Since then, this exchange of information on chemicals that are expected to occur in the environment, along with the accompanying expert knowledge and references, has become a valuable knowledge base for “suspect screening” lists. The NORMAN-SLE now serves as a FAIR (Findable, Accessible, Interoperable, Reusable) chemical information resource worldwide.
Results: The NORMAN-SLE contains 99 separate suspect list collections (as of May 2022) from over 70 contributors around the world, totalling over 100,000 unique substances. The substance classes include per- and polyfluoroalkyl substances (PFAS), pharmaceuticals, pesticides, natural toxins, high production volume substances covered under the European REACH regulation (EC: 1272/2008), priority contaminants of emerging concern (CECs) and regulatory lists from NORMAN partners. Several lists focus on transformation products (TPs) and complex features detected in the environment with various levels of provenance and structural information. Each list is available for separate download. The merged, curated collection is also available as the NORMAN Substance Database (NORMAN SusDat). Both the NORMAN-SLE and NORMAN SusDat are integrated within the NORMAN Database System (NDS). The individual NORMAN-SLE lists receive digital object identifiers (DOIs) and traceable versioning via a Zenodo community (https://zenodo.org/communities/norman-sle), with a total of > 40,000 unique views, > 50,000 unique downloads and 40 citations (May 2022). NORMAN-SLE content is progressively integrated into large open chemical databases such as PubChem (https://pubchem.ncbi.nlm.nih.gov/) and the US EPA’s CompTox Chemicals Dashboard (https://comptox.epa.gov/dashboard/), enabling further access to these lists, along with the additional functionality and calculated properties these resources offer. PubChem has also integrated significant annotation content from the NORMAN-SLE, including a classification browser (https://pubchem.ncbi.nlm.nih.gov/classification/#hid=101).
Conclusions: The NORMAN-SLE offers a specialized service for hosting suspect screening lists of relevance for the environmental community in an open, FAIR manner that allows integration with other major chemical resources. These efforts foster the exchange of information between scientists and regulators, supporting the paradigm shift to the “one substance, one assessment” approach. New submissions are welcome via the contacts provided on the NORMAN-SLE website (https://www.norman-network.com/nds/SLE/).

Microbial life in natural biofilms is dominated by prokaryotes and microscopic eukaryotes living in dense association. In stream ecosystems, microbial biofilms influence primary production, elemental cycles, food web interactions as well as water quality. Understanding how biofilm communities respond to anthropogenic impacts, such as wastewater treatment plant (WWTP) effluent, is important given the key role of biofilms in stream ecosystem function. Here, we implemented 16S and 18S rRNA gene sequencing of stream biofilms upstream (US) and downstream (DS) of WWTP effluents in four Swiss streams to test how bacterial and eukaryotic communities respond to wastewater constituents. Stream biofilm composition was strongly affected by geographic location – particularly for bacteria. However, the abundance of certain microbial community members was related to micropollutants in the wastewater – among bacteria, micropollutant-associated members were found e.g. in Alphaproteobacteria, and among eukaryotes e.g. in Bacillariophyta (algal diatoms). This study corroborates several previously characterized responses (e.g. as seen in diatoms), but also reveals previously unknown community responses – such as seen in Alphaproteobacteria. This study advances our understanding of the ecological impact of the current wastewater treatment practices and provides information about potential new marker organisms to assess ecological change in stream biofilms.

Advances in medicine and biotechnology rely on a deep understanding of biological processes. Despite the increasingly available types and amounts of omics data, significant knowledge gaps remain, with current approaches to identify and curate missing annotations being limited to a set of already known reactions. Here, we introduce Network Integrated Computational Explorer for Gap Annotation of Metabolism (NICEgame), a workflow to identify and curate nonannotated metabolic functions in genomes using the ATLAS of Biochemistry and genome-scale metabolic models (GEMs). To resolve gaps in GEMs, NICEgame provides alternative sets of known and hypothetical reactions, assesses their thermodynamic feasibility, and suggests candidate genes to catalyze these reactions. We identified metabolic gaps and applied NICEgame in the latest GEM of Escherichia coli, iML1515, and enhanced the E. coli genome annotation by resolving 47% of these gaps. NICEgame, applicable to any GEM and functioning from open-source software, should thus enhance all GEM-based predictions and subsequent biotechnological and biomedical applications.

Wicked problems exceed traditional sectoral and jurisdictional boundaries and involve multiple actors as stakeholders, victims, and culprits. Wicked problems inherently feature uncertainty and knowledge gaps. Science plays a crucial role in generating evidence for solving these problems and contributing to societal transformation. However, researchers may perceive and study problems detached from practitioners' perceptions of the world. We use the example of smallholder pesticide management in Uganda and the three types of knowledge approach, a framework borrowed from transdisciplinary research, to disentangle knowledge gaps. To identify these gaps, we integrated and co-produce knowledge in a two-day participatory workshop applying design thinking. Our results show, that a transition towards safe pesticide management depends on changes in the system, such as a revision and implementation of exiting regulation or professionalization of agro-dealers. Furthermore, this transition is only possible if interventions address target groups beyond the individual farmers (e.g. agro-dealers or district government officials). Compared to existing academic knowledge, co-produced knowledge provides a broader systemic perspective and yields more fine grained insights about potential new pathways. This investigation confirms, that practitioners’ knowledge is more fine-grained and detailed, thus exemplifying how knowledge integration is essential to avoid a gap between what researchers investigate and what practitioners need.

During dry periods, stream discharge in vegetated catchments can naturally fluctuate up to 10% daily. Despite intensive efforts put in observing and interpreting diel fluctuations of stream discharge across a range of instrumented natural catchments worldwide, the capability of state-of-the-art hydrological models to reproduce and explain such processes has rarely been tested. Here, we used CATHY, a physics-based integrated surface-subsurface hydrological model (ISSHM), to simulate the stream discharge in a small tile-drained agricultural catchment in Switzerland, where streamflow diel fluctuations appeared in dry periods. The model was able to satisfactorily reproduce the measured stream discharge, including the observed diel fluctuations. Next, we designed and simulated a series of modelling scenarios aimed to disentangle the processes contributing to the diel fluctuations in stream discharge. These scenarios revealed the predominant role of evapotranspiration (ET) in the establishment of diel fluctuations in stream discharge. Irrigation sustained the baseflow in dry periods and caused short-living streamflow peaks, which did not directly cause, but superposed to, diel patterns. Vegetation rooting depths and response to oxygen stress in the root zone slightly affected the amplitude of the streamflow diel fluctuations, while changes in the saturated hydraulic conductivity driven by diel soil temperature fluctuations caused an amplitude of the streamflow diel signal 10 times smaller compared to the one caused by ET. Our study demonstrates that ISSHMs such as CATHY can provide high-fidelity spatially-distributed dynamic simulations of evapotranspiration and irrigation fluxes, as well as soil moisture and groundwater flows, enhancing our understanding of the role of these hydrological processes during dry periods and thus making these models useful tools for a more sustainable management of agricultural catchments.

Um mit gezielten Massnahmen die Einträge von Pflanzenschutzmitteln in Oberflächengewässern zu verringern, müssen die relevanten Quellen und Eintragspfade bekannt sein. Dazu bestehen immer noch wichtige Wissenslücken. Deshalb wurde untersucht, welche Schlussfolgerungen aus zwölf bestehenden Monitoringstudien hinsichtlich der relevanten Transportwege gezogen werden können und was sich daraus für die Planung zukünftiger Studien und Projekte ableiten lässt.

Within the natural products field there is an increasing emphasis on the study of compounds from microbial sources. This has been fuelled by interest in the central role that microorganisms play in mediating both interspecies interactions and host-microbe relationships. To support the study of natural products chemistry produced by microorganisms we released the Natural Products Atlas, a database of known microbial natural products structures, in 2019. This paper reports the release of a new version of the database which includes a full RESTful application programming interface (API), a new website framework, and an expanded database that includes 8128 new compounds, bringing the total to 32 552. In addition to these structural and content changes we have added full taxonomic descriptions for all microbial taxa and have added chemical ontology terms from both NP Classifier and ClassyFire. We have also performed manual curation to review all entries with incomplete configurational assignments and have integrated data from external resources, including CyanoMetDB. Finally, we have improved the user experience by updating the Overview dashboard and creating a dashboard for taxonomic origin. The database can be accessed via the new interactive website at https://www.npatlas.org.

There is an increasing need for comparable and harmonized retention times (tR) in liquid chromatography (LC) among different laboratories, to provide supplementary evidence for the identity of compounds in high-resolution mass spectrometry (HRMS)-based suspect and nontarget screening investigations. In this study, a rigorously tested, flexible, and less system-dependent unified retention time index (RTI) approach for LC is presented, based on the calibration of the elution pattern. Two sets of 18 calibrants were selected for each of ESI+ and ESI-based on the maximum overlap with the retention times and chemical similarity indices from a total set of 2123 compounds. The resulting calibration set, with RTI set to range between 1 and 1000, was proposed as the most appropriate RTI system after rigorous evaluation, coordinated by the NORMAN network. The validation of the proposed RTI system was done externally on different instrumentation and LC conditions. The RTI can also be used to check the reproducibility and quality of LC conditions. Two quantitative structure-retention relationship (QSRR)-based models were built based on the developed RTI systems, which assist in the removal of false-positive annotations. The applicability domains of the QSRR models allowed completing the identification process with higher confidence for substances within the domain, while indicating those substances for which results should be treated with caution. The proposed RTI system was used to improve confidence in suspect and nontarget screening and increase the comparability between laboratories as demonstrated for two examples. All RTI-related calculations can be performed online at http://rti.chem.uoa.gr/.

Small streams in catchments with agricultural land use are at high risk of diffuse pollution by herbicides. Fast transport processes can cause concentration peaks that exceed regulatory requirements. These processes have a high spatio-temporal variability and data characterizing their occurrence is often sparse. For this reason, such systems show a stochastic behavior at the resolution we observe them (same input and initial conditions lead to different output). Realistic model representations should acknowledge this pronounced apparent intrinsic stochasticity. However, a deterministic description of the physical and chemical processes at the catchment scale is state of the art in research and practice. We explore the potential of stochastic process formulations in combination with the Bayesian learning paradigm to (a) improve the quantification of the uncertainty of conceptual catchment-scale pesticide transport models and (b) gain new mechanistic insights about the system by interpreting the temporal evolution of the stochastic processes. This is done with the help of a framework for time-varying stochastic parameters. Thereby, we find that (a) the stochastic process formulation can lead to a more realistic characterization of the uncertainty of internal states and model output compared to the deterministic one, and that (b) the temporal dynamics of parameters resulting from the inference can highlight model deficits (and inspire improvements) such as a better sustained baseflow in dry periods. We also identify two key challenges: numerical difficulties in sampling the posterior and the question of where to introduce and how to constrain the additional degrees of freedom such that they are not misused.

Methods are needed to verify that attenuation of energetic compounds, such as trinitrotoluene (TNT) and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX), in groundwater is occurring, and it must also be possible to verify that strategies to enhance degradation processes are having the desired effects. This document provides a primer on sample collection, processing, and laboratory methods for compound specific isotope analysis (CSIA)methods for nitro explosives. Data evaluation, analysis, and interpretation for both single and dual isotope fractionation is presented. Two case studies are used to demonstrate the procedures for using CSIA to support the identification of the relevant reaction process and calculate the extent of which reaction, versus dilution or sorption, is leading to decreases in concentration.

In this work, acidic oxidation in a biphasic water/octanol system was applied to mitigate condensation and increase the yields of high-value aromatic monomers from Kraft lignin, a waste product, and an industrially relevant feedstock. Biphasic depolymerization (BPD) is a one-pot multistep technique in which target molecules are protected from degradation in the reactive aqueous milieu (necessary for the cleavage of β-O-4 bonds as well as the recalcitrant high dissociation energy carbon-carbon moieties) by an in situ transfer of the products into the protective octanol phase. The results have shown overall monomeric yields up to 13 wt % and yields of vanillin up to 7 wt % (FeCl3 as a catalyst and O2 as an oxidant), significantly outperforming the previously reported monophasic and biphasic outcomes for Kraft lignin. In addition to increased yields, BPD also resulted in an expansion of the product pool from few molecules typically reported in the literature to 64 aromatic monomers detected at various concentration levels in the current work. The identified aromatic alcohols, carbonyl compounds, and carboxylic acids incorporating varying side chain lengths show potential as green and sustainable replacements to BTX (benzene, toluene, and xylene) petrochemicals. BPD as a process coupled mild conversion conditions, cheap oxidants, and common homogeneous catalysts with an in situ separation of the products from the reaction mixture to yield excellent results in converting industrial waste and recalcitrant feedstocks into value.

Wastewater treatment plants (WWTPs) play an important role in retaining organic matter and nutrients but to a lesser extent micropollutants. Therefore, treated wastewater is recognized as a major source of multiple stressors, including complex mixtures of micropollutants. These can potentially affect microbial communities in the receiving water bodies and the ecological functions they provide. In this study, we evaluated in flow-through channels the consequences of an exposure to a mixture of stream water and different percentages of urban WWTP effluent, ranging from 0% to 80%, on the microbial diversity and function of periphyton communities. Assuming that micropollutants exert a selective pressure for tolerant microorganisms within communities, we further examined the periphyton sensitivity to a micropollutant mixture extracted from passive samplers that were immersed in the wastewater effluent. As well, micropollutants in water and in periphyton were comprehensively quantified. Our results show that micropollutants detected in periphyton differed from those found in water, both in term of concentration and composition. Especially photosystem II inhibitors accumulated in periphyton more than other pesticides. Although effects of other substances cannot be excluded, this accumulation may have contributed to the observed higher tolerance of phototrophic communities to micropollutants upon exposure to 30% and 80% of wastewater. On the contrary, no difference in tolerance was observed for heterotrophic communities. Exposure to the gradient of wastewater led to structural differences in both prokaryotic and eukaryotic communities. For instance, the relative abundance of cyanobacteria was higher with increasing percentage of wastewater effluent, whereas the opposite was observed for diatoms. Such results could indicate that differences in community structure do not necessarily lead to higher tolerance. This highlights the need to consider other wastewater constituents such as nutrients and wastewater-derived microorganisms that can modulate community structure and tolerance. By using engineered flow-through channels that mimic to some extent the required field conditions for the development of tolerance in periphyton, our study constitutes a base to investigate the mechanisms underlying the increased tolerance, such as the potential role of microorganisms originating from wastewater effluents, and different treatment options to reduce the micropollutant load in effluents.

For most micropollutants (MPs) present in surface waters, such as pesticides and pharmaceuticals, the contribution of biotransformation to their overall removal from lake ecosystems is largely unknown. This study aims at empirically determining the biotransformation rate constants for 35 MPs at different periods of the year and depths of a meso-eutrophic lake. We then tested statistically the association of environmental parameters and microbial community composition with the biotransformation rate constants obtained. Biotransformation was observed for 14 out of 35 studied MPs for at least one sampling time. Large variations in biotransformation rate constants were observed over the seasons and between compounds. Overall, the transformation of MPs was mostly influenced by the lake's temperature, phytoplankton density and bacterial diversity. However, some individual MPs were not following the general trend or association with microorganism biomass. The antidepressant mianserin, for instance, was transformed in all experiments and depths, but did not show any relationship with measured environmental parameters, suggesting the importance of specific microorganisms in its transformation. The results presented here contribute to our understanding of the fate of MPs in surface waters and thus support improved risk assessment of contaminants in the environment.

Bedrock rivers represent a hydrogeological environment in which surface water flows along an exposed bedrock surface. Studies of hyporheic exchange have exclusively involved rivers composed of unconsolidated fluvial sediments, leaving a critical knowledge gap. This study evaluates the conditions that could support bedform-scale hyporheic exchange within a fractured sedimentary bedrock river based on field data collected near Guelph, Canada. Hyporheic exchange at the bedform-scale was evaluated by numerically modeling the migration of a conservative solute tracer through a bedrock riverbed within a two-dimensional vertical cross-section along the flow direction. A stochastic discrete fracture-matrix framework was developed to represent measured subsurface fractured bedrock properties, producing a probabilistic distribution of potential hyporheic exchange pathways. Flow and transport model results indicate that: (a) bedform-scale hyporheic exchange within a bedrock river exists with high fluid flow velocities in fractures, yet long solute residence times due to diffusion across the fracture-matrix boundary; (b) the coincidence of fractures and hydrodynamic head gradients across the riverbed controls the spatial extent of bedform-scale hyporheic exchange; and (c) the potential variability in hyporheic exchange residence time is large (i.e., tens of years) due to the inherent variability in facture network properties. Our field-based numerical study indicates that the average (median) residence time may not be a good proxy for the potential natural attenuation capacity of a fractured sedimentary bedrock riverbed and that hyporheic exchange has the potential to emplace surface water contaminants within the fractured porous rock matrix that could become a long-term source of trace contaminants.

In this study, the SWAT hydrological model was used to estimate the sediment yields in the principal drainage basins of the Congo River Basin. The model was run for the 2000–2012 period and calibrated using measured values obtained at the basins principal gauging station that controls 98% of the basin area. Sediment yield rates of 4.01, 5.91, 7.88 and 8.68 t km⁻² yr⁻¹ were estimated for the areas upstream of the Ubangi at Bangui, Sangha at Ouesso, Lualaba at Kisangani, and Kasai at Kuto-Moke, respectively—the first three of which supply the Cuvette Centrale. The loads contributed into the Cuvette Centrale by eight tributaries were estimated to be worth 0.04, 0.07, 0.09, 0.18, 0.94, 1.50, 1.60, and 26.98 × 10⁶ t yr⁻¹ from the Likouala Mossaka at Makoua, Likouala aux Herbes at Botouali, Kouyou at Linnegue, Alima at Tchikapika, Sangha at Ouesso, Ubangi at Mongoumba, Ruki at Bokuma and Congo at Mbandaka, respectively. The upper Congo supplies up to 85% of the fluxes in the Cuvette Centrale, with the Ubangi and the Ruki contributing approximately 5% each. The Cuvette Centrale acts like a big sink trapping up to 23 megatons of sediment produced upstream (75%) annually.

Micropollutants are ubiquitously found in natural surface waters and pose a potential risk to aquatic organisms. Stream biofilms, consisting of bacteria, algae and other microorganisms potentially contribute to bioremediating aquatic environments by biotransforming xenobiotic substances. When investigating the potential of stream biofilms to remove micropollutants from the water column, it is important to distinguish between different fate processes, such as biotransformation, passive sorption and active bioaccumulation. However, due to the complex nature of the biofilm community and its extracellular matrix, this task is often difficult. In this study, we combined biotransformation experiments involving natural stream biofilms collected up- and downstream of wastewater treatment plant outfalls with the QuEChERS extraction method to distinguish between the different fate processes. The QuEChERS extraction proved to be a suitable method for a broad range of micropollutants (> 80% of the investigated compounds). We found that 31 out of 63 compounds were biotransformed by the biofilms, with the majority being substitution-type biotransformations, and that downstream biofilms have an increased biotransformation potential towards specific wastewater-relevant micropollutants. Overall, using the experimental and analytical strategy developed, stream biofilms were demonstrated to have a broad inherent micropollutant biotransformation potential, and to thus contribute to bioremediation and improving ecosystem health.

Studies suggest that acute pesticide poisonings (APP) may be linked with long-term neurological effects. To examine long-term neurological and psychological distress symptoms associated with having experienced an APP, we conducted a cross-sectional study of 300 conventional and organic smallholder farmers from Zarcero County, Costa Rica, May–August 2016. We collected self-reported data on sociodemographic characteristics, occupational history, pesticide exposure, APPs, neurological and psychological distress symptoms (using the Brief Symptom Inventory (BSI)). Adjusted logistic regression models were fit. A total of 14% of the farmers (98% male) reported experiencing at least one APP during their lifetime. Self-reported APP was associated with neurological symptoms during the 12 months prior to interview (e.g., fainting (Odds Ratio: 7.48, 95% Confidence Interval: 1.83, 30.74), shaking hands (3.50; 1.60, 7.60), numbness/tingling in hands or feet (3.23; 1.66, 6.32), insomnia (2.53; 1.34, 4.79), accelerated heartrate (2.42; 1.03, 5.47), dizziness (2.38; 1.19, 4.72), increased irritability/anger (2.37; 1.23, 4.55), low energy (2.33; 1.23, 4.46), and difficulty concentrating (2.01; 1.05, 3.85)). Farmers who reported an APP in the ten years prior to interview experienced increased odds of abnormal BSI scores for hostility (4.51; 1.16, 17.70) and paranoid ideation (3.76; 0.99, 18.18). Having experienced an APP may be associated with long-term neurological and psychological distress symptoms.

Complex microbial communities in environmental systems play a key role in the detoxification of chemical contaminants by transforming them into less active metabolites or by complete mineralization. Biotransformation, i.e., transformation by microbes, is well understood for a number of priority pollutants, but a similar level of understanding is lacking for many emerging contaminants encountered at low concentrations and in complex mixtures across natural and engineered systems. Any advanced approaches aiming to reduce environmental exposure to such contaminants (e.g., novel engineered biological water treatment systems, design of readily degradable chemicals, or improved regulatory assessment strategies to determine contaminant persistence a priori) will depend on understanding the causal links among contaminant removal, the key driving agents of biotransformation at low concentrations (i.e., relevant microbes and their metabolic activities), and how their presence and activity depend on environmental conditions. In this Perspective, we present the current understanding and recent methodological advances that can help to identify such links, even in complex environmental microbiomes and for contaminants present at low concentrations in complex chemical mixtures. We discuss the ensuing insights into contaminant biotransformation across varying environments and conditions and ask how much closer we have come to designing improved approaches to reducing environmental exposure to contaminants.

Background: Bloom-forming cyanobacteria occur globally in aquatic environments. They produce diverse bioactive metabolites, some of which are known to be toxic. The most studied cyanobacterial toxins are microcystins, anatoxin, and cylindrospermopsin, yet more than 2000 bioactive metabolites have been identified to date. Data on the occurrence of cyanopeptides other than microcystins in surface waters are sparse.
Results: We used a high-performance liquid chromatography–high-resolution tandem mass spectrometry/tandem mass spectrometry (HPLC–HRMS/MS) method to analyse cyanotoxin and cyanopeptide profiles in raw drinking water collected from three freshwater reservoirs in the United Kingdom. A total of 8 cyanopeptides were identified and quantified using reference standards. A further 20 cyanopeptides were identified based on a suspect-screening procedure, with class-equivalent quantification. Samples from Ingbirchworth reservoir showed the highest total cyanopeptide concentrations, reaching 5.8, 61, and 0.8 µg/L in August, September, and October, respectively. Several classes of cyanopeptides were identified with anabaenopeptins, cyanopeptolins, and microcystins dominating in September with 37%, 36%, and 26%, respectively. Samples from Tophill Low reservoir reached 2.4 µg/L in September, but remained below 0.2 µg/L in other months. Samples from Embsay reservoir did not exceed 0.1 µg/L. At Ingbirchworth and Tophill Low, the maximum chlorophyll-a concentrations of 37 µg/L and 22 µg/L, respectively, and cyanobacterial count of 6 × 10⁴ cells/mL were observed at, or a few days after, peak cyanopeptide concentrations. These values exceed the World Health Organization’s guideline levels for relatively low probability of adverse health effects, which are defined as 10 µg/L chlorophyll-a and 2 × 10⁴ cells/mL.
Conclusions: This data is the first to present concentrations of anabaenopeptins, cyanopeptolins, aeruginosins, and microginins, along  with microcystins, in U.K. reservoirs. A better understanding of those cyanopeptides that are abundant in drinking water reservoirs can inform future monitoring and studies on abatement efficiency during water treatment.

The exposure of ecologically critical invertebrate species to biologically active pharmaceuticals poses a serious risk to the aquatic ecosystem. Yet, the fate and toxic effects of pharmaceuticals on these nontarget aquatic invertebrates and the underlying mechanisms are poorly studied. Herein, we investigated the toxicokinetic (TK) processes (i.e., uptake, biotransformation, and elimination) of the pharmaceutical diclofenac and its biotransformation in the freshwater invertebrate Hyalella azteca. We further employed mass spectrometry-based metabolomics to assess the toxic effects of diclofenac on the metabolic functions of H. azteca exposed to environmentally relevant concentrations (10 and 100 μg/L). The TK results showed a quick uptake of diclofenac by H. azteca (maximum internal concentration of 1.9 μmol/kg) and rapid formation of the conjugate diclofenac taurine (maximum internal concentration of 80.6 μmol/kg), indicating over 40 times higher accumulation of diclofenac taurine than that of diclofenac in H. azteca. Depuration kinetics demonstrated that the elimination of diclofenac taurine was 64 times slower than diclofenac in H. azteca. Metabolomics results suggested that diclofenac inhibited prostaglandin synthesis and affected the carnitine shuttle pathway at environmentally relevant concentrations. These findings shed light on the significance of the TK process of diclofenac, especially the formation of diclofenac taurine, as well as the sublethal effects of diclofenac on the bulk metabolome of H. azteca. Combining the TK processes and metabolomics provides complementary insights and thus a better mechanistic understanding of the effects of diclofenac in aquatic invertebrates.

Background: Multiple epidemiological studies have shown that exposure to single pesticide active ingredients or chemical groups is associated with adverse neurobehavioral outcomes in farmers. In agriculture, exposure to multiple pesticide active ingredients is the rule, rather than exception. Therefore, occupational studies on neurobehavioral effects of pesticides should account for potential co-exposure confounding.
Methods: We conducted a cross-sectional study of 288 Ugandan smallholder farmers between September and December 2017. We collected data on self-reported use of pesticide products during the 12 months prior to survey and estimated yearly exposure-intensity scores for 14 pesticide active ingredients using a semi-quantitative exposure algorithm. We administered 11 neurobehavioral tests to assess five neurobehavioral domains. We implemented a Bayesian Model-Averaging (BMA) approach to examine the association between exposure to multiple pesticides and neurobehavioral outcomes, while accounting for multiple testing. We applied two levels of inference to determine (1) which neurobehavioral outcomes were associated with overall pesticide exposure (marginal inclusion probability (MIP) for covariate-only models <0.5) and (2) which specific pesticide active ingredients were associated with these outcomes (MIP for models where active ingredient was included >0.5).
Results: Seventy-two percent of farmers reported use of pesticide products that contained at least one of 14 active ingredients, while the applicators used in median three different active ingredients (interquartile range (IQR) 4) in the 12 months prior to the study. The most widely used active ingredients were glyphosate (79%), cypermethrin (60%), and mancozeb (55%). We found that overall pesticide exposure was associated with impaired visual memory (Benton Visual Retention Test (BVRT)), language (semantic verbal fluency test), perceptual-motor function (Finger tapping test), and complex attention problems (Trail making A test and digit symbol test). However, when we looked at the associations for individual active ingredients, we only observed a positive association between glyphosate exposure and impaired visual memory (-0.103 [95% Bayesian Credible Interval (BCI)] [-0.24, 0] units in BVRT scores per interquartile range (IQR) increase in annual exposure to glyphosate, relative to a median [IQR] of 6 [3] units in BVRT across the entire study population).
Conclusions: We found that overall pesticide exposure was associated with several neurobehavioral outcome variables. However, when we examined individual pesticide active ingredients, we observed predominantly null associations, except for a positive association between glyphosate exposure and impaired visual memory. Additional epidemiologic studies are needed to evaluate glyphosate’s neurotoxicity, while accounting for co-pollutant confounding.

Ozonation is increasingly applied in water and wastewater treatment for the abatement of micropollutants (MPs). However, the transformation products formed during ozonation (OTPs) and their fate in biological or sorptive post-treatments is largely unknown. In this project, a high-throughput approach, combining laboratory ozonation experiments and detection by liquid chromatography high-resolution mass spectrometry (LC-HR-MS/MS), was developed and applied to identify OTPs formed during ozonation of wastewater effluent for a large number of relevant MPs (total 87). For the laboratory ozonation experiments, a simplified experimental solution, consisting of surrogate organic matter (methanol and acetate), was created, which produced ozonation conditions similar to realistic conditions in terms of ozone and hydroxyl radical exposures. The 87 selected parent MPs were divided into 19 mixtures, which enabled the identification of OTPs with an optimized number of experiments. The following two approaches were considered to identify OTPs. (1) A screening of LC-HR-MS signal formation in these experiments was performed and revealed a list of 1749 potential OTP candidate signals associated to 70 parent MPs. This list can be used in future suspect screening studies. (2) A screening was performed for signals that were formed in both batch experiments and in samples of wastewater treatment plants (WWTPs). This second approach was ultimately more time-efficient and was applied to four different WWTPs with ozonation (specific ozone doses in the range 0.23-0.55 gO₃/gDOC), leading to the identification of 84 relevant OTPs of 40 parent MPs in wastewater effluent. Chemical structures could be proposed for 83 OTPs through the interpretation of MS/MS spectra and expert knowledge in ozone chemistry. Forty-eight OTPs (58%) have not been reported previously. The fate of the verified OTPs was studied in different post-treatment steps. During sand filtration, 87-89% of the OTPs were stable. In granular activated carbon (GAC) filters, OTPs were abated with decreasing efficiency with increasing run times of the filters. For example, in a GAC filter with 16,000 bed volumes, 53% of the OTPs were abated, while in a GAC filter with 35,000 bed volumes, 40% of the OTPs were abated. The highest abatement (87% of OTPs) was observed when 13 mg/L powdered activated carbon (PAC) was dosed onto a sand filter.

Micropollutants (MP) with varying ozone-reactive moieties were spiked to lake water in the influent of a drinking water pilot plant consisting of an ozonation followed by a biological sand filtration. During ozonation, 227 transformation products (OTPs) from 39 of the spiked 51 MPs were detected after solid phase extraction by liquid chromatography high-resolution mass spectrometry (LC-HRMS/MS). Based on the MS/MS data, tentative molecular structures are proposed. Reaction mechanisms for the formation of a large number of OTPs are suggested by combination of the kinetics of formation and abatement and state-of-the-art knowledge on ozone and hydroxyl radical chemistry. OTPs forming as primary or higher generation products from the oxidation of MPs could be differentiated. However, some expected products from the reactions of ozone with activated aromatic compounds and olefins were not detected with the applied analytical procedure. 187 OTPs were present in the sand filtration in sufficiently high concentrations to elucidate their fate in this treatment step. 35 of these OTPs (19%) were abated in the sand filtration step, most likely due to biodegradation. Only 24 (13%) of the OTPs were abated more efficiently than the parent compounds, with a dependency on the functional group of the parent MPs and OTPs. Overall, this study provides evidence, that the common assumption that OTPs are easily abated in biological post-treatment is not generally valid. Nevertheless, it is unknown how the OTPs, which escaped detection, would have behaved in the biological post-treatment.

In einer Pilotanlage zur Aufbereitung von Seewasser wurden in der Ozonungsstufe 227 Transformationsprodukte detektiert, die aus 39 der insgesamt 51 dem Seewasser zugegebenen Spurenstoffe entstanden waren. Weniger als 20% dieser Transformationsprodukte waren in der nachgeschalteten biologischen Sandfiltration abbaubar. Zudem waren insgesamt nur 13% der im Sandfilter nachgewiesenen Transformationsprodukte besser abbaubar als die Ausgangssubstanzen.

Les pesticides sont avant tout utilisés dans l’agriculture, mais aussi en milieu urbain et dans d'autres domaines. Sous forme de produits phytosanitaires, ils réduisent les pertes de rendement et de qualité dues aux organismes nuisibles. Toutefois, l'emploi actuel de pesticides porte toutefois un préjudice considérable à l'environnement et en particulier à la biodiversité. En dépit de certaines lacunes, les répercussions indésirables sont bien documentées. La présente fiche d'information aborde spécifiquement les incidences de l'usage de pesticides sur l'environnement, la biodiversité et les services écosystémiques et propose des pistes d'action possibles pour la politique et la société, visant à réduire les effets indésirables. Elle ne thématise pas, par exemple, l'importance des pesticides pour la production agricole ou les incidences sur la santé humaine.

Pestizide werden primär in der Landwirtschaft, aber auch im Siedlungsraum und in weiteren Bereichen eingesetzt. Als Pflanzenschutzmittel vermindern sie Ertrags- oder Qualitätseinbussen durch Schadorganismen. Der heutige Pestizideinsatz belastet aber die Umwelt und insbesondere die Biodiversität beträchtlich. Trotz Wissenslücken sind die unerwünschten Auswirkungen gut dokumentiert. Das Faktenblatt nimmt spezifisch die Auswirkungen von Pestiziden auf Umwelt, Biodiversität und Ökosystemleistungen in den Blick und zeigt mögliche Handlungsansätze für Politik und Gesellschaft zur Reduktion der unerwünschten Nebenwirkungen des Pestizideinsatzes auf. Nicht thematisiert werden im Faktenblatt etwa die Bedeutung von Pestiziden für die landwirtschaftliche Produktion oder die Auswirkungen auf die menschliche Gesundheit.

Toxic plant secondary metabolites (PSMs), so-called phytotoxins, occur widely in plant species. Many of these phytotoxins have similar mobility, persistence, and toxicity properties in the environment as anthropogenic micropollutants, which increasingly contaminate surface waters. Although recent case studies have shown the aquatic relevance of phytotoxins, the overall exposure remains unknown. Therefore, we performed a detailed occurrence analysis covering 134 phytotoxins from 27 PSM classes. Water samples from seven small Swiss streams with catchment areas from 1.7 to 23 km² and varying land uses were gathered over several months to investigate seasonal impacts. They were complemented with samples from different biogeographical regions to cover variations in vegetation. A broad SPE-LC-HRMS/MS method was applied with limits of detection below 5 ng/L for over 80% of the 134 included phytotoxins. In total, we confirmed 39 phytotoxins belonging to 13 PSM classes, which corresponds to almost 30% of all included phytotoxins. Several alkaloids were regularly detected in the low ng/L-range, with average detection frequencies of 21%. This is consistent with the previously estimated persistence and mobility properties that indicated a high contamination potential. Coumarins were previously predicted to be unstable, however, detection frequencies were around 89%, and maximal concentrations up to 90 ng/L were measured for fraxetin produced by various trees. Overall, rainy weather conditions at full vegetation led to the highest total phytotoxin concentrations, which might potentially be most critical for aquatic organisms.

Pyrrolizidine alkaloids (PAs) are found to be toxic pollutants emitted into the environment by numerous plant species, resulting in contamination. In this article, we investigate the occurrence of PAs in the aquatic environment of small Swiss streams combining two different approaches. Pyrrolizidine alkaloids (PAs) are toxic secondary metabolites produced by numerous plant species. Although they were classified as persistent and mobile and found to be emitted into the environment, their occurrence in surface waters is largely unknown. Therefore, we performed a retrospective data analysis of two extensive HRMS campaigns each covering five small streams in Switzerland over the growing season. All sites were contaminated with up to 12 individual PAs and temporal detection frequencies between 36 and 87%. Individual PAs were in the low ng/L range, but rain-induced maximal total PA concentrations reached almost 100 ng/L in late spring and summer. Through PA patterns in water and plants, several species were tentatively identified as the source of contamination, with Senecio spp. and Echium vulgare being the most important. Additionally, two streams were monitored, and PAs were quantified with a newly developed, faster, and more sensitive LC−MS/MS method to distinguish different plant-based and indirect human PA sources. A distinctly different PA fingerprint in aqueous plant extracts pointed to invasive Senecio inaequidens as the main source of the surface water contamination at these sites. Results indicate that PA loads may increase if invasive species are sufficiently abundant.

Introduction and aim Exposure to some insecticides may cause airway obstruction, but existing evidence is limited by cross-sectional designs and inadequate confounder control. We investigated the relation between organophosphate and carbamate insecticides and pulmonary function in a prospective study accounting for important confounders.
Methods In a cohort of 364 smallholder farmers in Uganda (69% women), participants underwent pre-bronchodilator spirometry at baseline (September/October 2018) and at two follow-up visits (November/December 2018 and January/February 2019). Exposure to carbamate and organophosphate insecticides was assessed using haemoglobin-adjusted erythrocyte acetylcholinesterase (AChE/Hb). Less than 3% of participants were lost to follow-up. We calculated Z-scores for FEV₁, FVC and FEV₁/FVC using the Global Lung Function Initiative equations. Data were analysed in linear mixed and fixed effect models accounting for family relationships and repeated measures of exposure and outcome.
Results Low AChE/Hb was significantly associated with low FEV₁ Z-score in both unadjusted and adjusted analyses. Compared with individuals with AChE/Hb 25.90 U/g (50^th percentile, reference), those with lower AChE/Hb 24.50 U/g (35^th percentile) had mean FEV₁ Z-score 0.045 (0.003 to 0.087) lower, and persons with higher AChE/Hb 27.30 U/g (65^th percentile) had a mean FEV₁ Z-score 0.043 (-0.002 to 0.087) higher compared with the reference. Similar, but numerically smaller and statistically non-significant effects were seen for Z-scores of FVC and FEV₁/FVC.
Conclusion Exposure to organophosphate and carbamate insecticides may lead to lung function decline. Our results add to the growing evidence of health effects in relation to exposure to organophosphate and carbamate insecticides, underlining the importance of minimising exposure.

Objective
Evaluate the accuracy and precision of the copd-6 mini-spirometer for FEV₁ in a rural Ugandan population.

Methods
In a cross-sectional study, 171 smallholder farmers performed spirometry with copd-6, and a diagnostic-quality spirometer.

Results and discussion
The copd-6 underestimated FEV₁ at low flows and overestimated FEV₁ at high flows. Across all participants, the device slightly overestimated FEV₁ by 0.04 [0.02; 0.06] L. Calibration data showed similar patterns.

Conclusion
The copd-6 could be considered as an affordable tool for research on lung function impairment in resource-constrained settings. However, further validation in a study population with obstructive lung disease is needed.

Due to the low environmental concentrations and substance-specific behavior of trace organic chemicals (TOrCs), general mechanistic insights can hardly be gained from biotransformation studies with individual substances. In this study, we aimed to identify prevalent enzymatic reactions under different redox and substrate conditions in a well-defined laboratory-scale column system. Biotransformation of chemicals in the first column, which was characterized by a relatively high biomass and rapid depletion of dissolved oxygen, was initiated by frequent and fast reactions such as the oxidation of alcohols or amide hydrolysis. Slower reactions, e.g., the initial dealkylation of primary and secondary amines, were inefficient in the first column and benefited from oxic and carbon-limited conditions prevalent in the second column system after reaeration. Moreover, several compound-specific reactions such as the cleavage of C-Cl and C-O bonds through substitution with glutathione occurred only under carbon-limited conditions. The link between system-specific TOrC removal and suggested initial reactions from the literature can serve as a starting point for a suspect screening of relevant enzymes for the biotransformation of TOrCs based on metagenomic or metatranscriptomic data. Complementary batch experiments with media from the columns confirmed the observed removal under carbon-rich conditions but revealed a limited reproducibility of microbial degradation under oligotrophic conditions.

Urban streams receive increasing loads of organic micropollutants from treated wastewaters. A comprehensive understanding of the in-stream fate of micropollutants is thus of high interest for water quality management. Bedforms induce pumping effects considerably contributing to whole stream hyporheic exchange and are hotspots of biogeochemical turnover processes. However, little is known about the transformation of micropollutants in such structures. In the present study, we set up recirculating flumes to examine the transformation of a set of micropollutants along single flowpaths in two triangular bedforms. We sampled porewater from four locations in the bedforms over 78 days and analysed the resulting concentration curves using the results of a hydrodynamic model in combination with a reactive transport model accounting for advection, dispersion, first-order removal and retardation. The four porewater sampling locations were positioned on individual flowpaths with median solute travel times ranging from 11.5 to 43.3 h as shown in a hydrodynamic model previously. Highest stability was estimated for hydrochlorothiazide on all flowpaths. Lowest detectable half-lives were estimated for sotalol (0.7 h) and sitagliptin (0.2 h) along the shortest flowpath. Also, venlafaxine, acesulfame, bezafibrate, irbesartan, valsartan, ibuprofen and naproxen displayed lower half-lives at shorter flowpaths in the first bedform. However, the behavior of many compounds in the second bedform deviated from expectations, where particularly transformation products, e.g. valsartan acid, showed high concentrations. Flowpath-specific behavior as observed for metformin or flume-specific behavior as observed for metoprolol acid, for instance, was attributed to potential small-scale or flume-scale heterogeneity of microbial community compositions, respectively. The results of the study indicate that the shallow hyporheic flow field and the small-scale heterogeneity of the microbial community are major controlling factors for the transformation of relevant micropollutants in river sediments.

Harmful cyanobacterial blooms, which frequently contain toxic secondary metabolites, are reported in aquatic environments around the world. More than two thousand cyanobacterial secondary metabolites have been reported from diverse sources over the past fifty years. A comprehensive, publically-accessible database detailing these secondary metabolites would facilitate research into their occurrence, functions and toxicological risks. To address this need we created CyanoMetDB, a highly curated, flat-file, openly-accessible database of cyanobacterial secondary metabolites collated from 850 peer-reviewed articles published between 1967 and 2020. CyanoMetDB contains 2010 cyanobacterial metabolites and 99 structurally related compounds. This has nearly doubled the number of entries with complete literature metadata and structural composition information compared to previously available open access databases. The dataset includes microcytsins, cyanopeptolins, other depsipeptides, anabaenopeptins, microginins, aeruginosins, cyclamides, cryptophycins, saxitoxins, spumigins, microviridins, and anatoxins among other metabolite classes. A comprehensive database dedicated to cyanobacterial secondary metabolites facilitates: (1) the detection and dereplication of known cyanobacterial toxins and secondary metabolites; (2) the identification of novel natural products from cyanobacteria; (3) research on biosynthesis of cyanobacterial secondary metabolites, including substructure searches; and (4) the investigation of their abundance, persistence, and toxicity in natural environments.

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.

Groundwater is a major drinking water resource but its quality with regard to organic micropollutants (MPs) is insufficiently assessed. Therefore, we aimed to investigate Swiss groundwater more comprehensively using liquid chromatography high-resolution tandem mass spectrometry (LC-HRMS/MS). First, samples from 60 sites were classified as having high or low urban or agricultural influence based on 498 target compounds associated with either urban or agricultural sources. Second, all LC-HRMS signals were related to their potential origin (urban, urban and agricultural, agricultural, or not classifiable) based on their occurrence and intensity in the classified samples. A considerable fraction of estimated concentrations associated with urban and/or agricultural sources could not be explained by the 139 detected targets. The most intense nontarget signals were automatically annotated with structure proposals using MetFrag and SIRIUS4/CSI:FingerID with a list of >988,000 compounds. Additionally, suspect screening was performed for 1162 compounds with predicted high groundwater mobility from primarily urban sources. Finally, 12 nontargets and 11 suspects were identified unequivocally (Level 1), while 17 further compounds were tentatively identified (Level 2a/3). amongst these were 13 pollutants thus far not reported in groundwater, such as: the industrial chemicals 2,5-dichlorobenzenesulfonic acid (19 detections, up to 100 ng L^-1), phenylphosponic acid (10 detections, up to 50 ng L^-1), triisopropanolamine borate (2 detections, up to 40 ng L^-1), O-des[2-aminoethyl]-O-carboxymethyl dehydroamlodipine, a transformation product (TP) of the blood pressure regulator amlodipine (17 detections), and the TP SYN542490 of the herbicide metolachlor (Level 3, 33 detections, estimated concentrations up to 100–500 ng L^-1). One monitoring site was far more contaminated than other sites based on estimated total concentrations of potential MPs, which was supported by the elucidation of site-specific nontarget signals such as the carcinogen chlorendic acid, and various naphthalenedisulfonic acids. Many compounds remained unknown, but overall, source related prioritisation proved an effective approach to support identification of compounds in groundwater.

Groundwater is a major drinking water resource, providing on average 50% of domestic water supply worldwide; in several European countries, this can even be as high as 70-100%. Although groundwater is better protected from anthropogenic influences than surface waters, a broad variety of chemicals used in agriculture, households, and industry have been detected in groundwater in the ngL^-1 to μgL^-1 range. Chemicals may enter groundwater via seepage after pesticide application to agricultural soils, from leaky sewers, or from the infiltration of wastewater-impacted surface waters. Natural attenuation processes such as sorption and (bio)transformation do not sufficiently retain polar, mobile, and persistent micropollutants (MPs) and may even lead to the formation of polar transformation products (TPs). Compared to the large number of chemicals potentially released into the aquatic environment, previous research and monitoring programs focused only on a few compounds so that groundwater quality with regard to organic MPs is largely unknown. [...]

Grundwasser ist eine wichtige Trinkwasserressource, welche weltweit 50% des Wasserbedarfs von Haushalten bereitstellt und in mehreren europäischen Ländern sogar 70-100%. Obwohl Grundwasser besser vor anthropogenen Einflüssen geschützt ist als Oberflächengewässer, wurden im Grundwasser zahlreiche Chemikalien aus der Landwirtschaft, Haushalten und Industrie im ngL^-1 bis μgL^-1 Bereich nachgewiesen. Chemikalien können mit dem Sickerwasser ins Grundwasser eingetragen werden infolge der Anwendung von Pestiziden auf  landwirtschaftlichen Böden, aufgrund undichter Abwasserkanäle oder durch die Infiltration von mit Abwasser belasteten Oberflächengewässern. Natürliche Prozesse wie Sorption und (Bio-) Transformation halten polare, mobile und persistente Spurenstoffe nicht ausreichend zurück und können sogar zur Bildung polarer Transformationsprodukte (TPs) führen. Im Vergleich zur grossen Zahl an Chemikalien, die möglicherweise in den Wasserkreislauf freigesetzt werden, umfassten bisherige Untersuchungen und Monitoringprogramme nur wenige Verbindungen, so dass die Grundwasserqualität in Bezug auf organische Spurenstoffe weitgehend unbekannt ist. [...]

A wide range of micropollutants can be monitored with non-targeted screening; however, the quantification of the newly discovered compounds is challenging. Transformation products (TPs) are especially problematic because analytical standards are rarely available. Here, we compared three quantification approaches for non-target compounds that do not require the availability of analytical standards. The comparison is based on a unique set of concentration data for 341 compounds, mainly pesticides, pharmaceuticals, and their TPs in 31 groundwater samples from Switzerland. The best accuracy was observed with the predicted ionization efficiency-based quantification, the mean error of concentration prediction for the groundwater samples was a factor of 1.8, and all of the 74 micropollutants detected in the groundwater were quantified with an error less than a factor of 10. The quantification of TPs with the parent compounds had significantly lower accuracy (mean error of a factor of 3.8) and could only be applied to a fraction of the detected compounds, while the mean performance (mean error of a factor of 3.2) of the closest eluting standard approach was similar to the parent compound approach.

This study simulates carbon dioxide (CO₂) sequestration in 300 major world river basins (about 70% of global surface area) through carbonates dissolution and silicate hydrolysis. For each river basin, the daily timescale impacts under the RCP 2.6 and RCP 8.5 climate scenarios were assessed relative to a historical baseline (1969–1999) using a cascade of models accounting for the hydrological evolution under climate change scenarios. Here we show that the global temporal evolution of the CO₂ uptake presents a general increase in the annual amount of CO₂ consumed from 0.247 ± 0.045 Pg C year⁻¹ to 0.261 and 0.273 ± 0.054 Pg C year⁻¹, respectively for RCP 2.6 and RCP 8.5. Despite showing a general increase in the global daily carbon sequestration, both climate scenarios show a decrease between June and August. Such projected changes have been mapped and evaluated against changes in hydrology, identifying hot spots and moments for the annual and seasonal periods.

River networks are one of the main routes by which the public could be exposed to environmental sources of antibiotic resistance, that may be introduced e.g. via treated wastewater. In this study, we applied a comprehensive integrated analysis encompassing mass-flow concepts, chemistry, bacterial plate counts, resistance gene quantification and shotgun metagenomics to track the fate of the resistome (collective antibiotic resistance genes (ARGs) in a microbial community) of treated wastewater in two Swiss rivers at the kilometer scale. The levels of certain ARGs and the class 1 integron integrase gene (intI1) commonly associated with anthropogenic sources of ARGs decreased quickly over short distances (2-2.5 km) downstream of wastewater discharge points. Mass-flow analysis based on conservative tracers suggested this decrease was attributable mainly to dilution but ARG loadings frequently also decreased (e.g., 55.0-98.5 % for ermB and tetW) over the longest studied distances (6.8 and 13.7 km downstream). Metagenomic analysis confirmed that ARG of wastewater-origin did not persist in rivers after 5 ∼ 6.8 km downstream distance. sul1 and intI1 levels and loadings were more variable and even increased sharply at 5 ∼ 6.8 km downstream distance on one occasion. While input from agriculture and in-situ positive selection pressure for organisms carrying ARGs cannot be excluded, in-system growth of biomass is a more probable explanation. The potential for direct human exposure to the resistome of wastewater-origin thus appeared to typically abate rapidly in the studied rivers. However, the riverine aquatic resistome was also dynamic, as evidenced by the increase of certain gene markers downstream, without obvious sources of anthropogenic contamination. This study provides new insight into drivers of riverine resistomes and pinpoints key monitoring targets indicative of where human sources and exposures are likely to be most acute.

Synthetic water-soluble polymeric materials are widely employed in e.g. cleaning detergents, personal care products, paints or textiles. Accordingly, these compounds reach sewage treatment plants and may enter receiving waters and the aquatic environment. Characteristically, these molecules show a polydisperse molecular weight distribution, comprising multiple repeating units, i.e. a homologous series (HS). Their analysis in environmentally relevant samples has received some attention over the last two decades, however, the majority of previous studies focused on surfactants and a molecular weight range <1000 Da. To capture a wider range on the mass versus polarity plane and extend towards less polar contaminants, a workflow was established using three different ionization strategies, namely conventional electrospray ionization, atmospheric pressure photoionization and atmospheric pressure chemical ionization. The data evaluation consisted of suspect screening of ca. 1200 suspect entries and a non-target screening of HS with pre-defined accurate mass differences using ca. 400 molecular formulas of repeating units of HS as input and repeating retention time shifts as HS indicator.
To study the fate of these water-soluble polymeric substances in the wastewater treatment process, the different stages, i.e. after primary and secondary clarifier, and after ozonation followed by sand filtration, were sampled at a Swiss wastewater treatment plant. Remaining with two different ionization interfaces, ESI and APPI, in both polarities, a non-targeted screening approach led to a total number of 146 HS (each with a minimum number of 4 members), with a molecular mass of up to 1200 detected in the final effluent. Of the 146 HS, ca 15% could be associated with suspect hits and approximately 25% with transformation products of suspects. Tentative characterization or probable chemical structure could be assigned to almost half of the findings. In positive ionization mode various sugar derivatives with differing side chains, for negative mode structures with sulfonic acids, could be characterized. The number of detected HS decreased significantly over the three treatment stages. For HS detectable also in the biological and oxidative treatment stages, a change in HS distribution towards to lower mass range was often observed.

Toxicology has been an active research field for many decades, with academic, industrial and government involvement. Modern omics and computational approaches are changing the field, from merely disease-specific observational models into target-specific predictive models. Traditionally, toxicology has strong links with other fields such as biology, chemistry, pharmacology and medicine. With the rise of synthetic and new engineered materials, alongside ongoing prioritisation needs in chemical risk assessment for existing chemicals, early predictive evaluations are becoming of utmost importance to both scientific and regulatory purposes. ELIXIR is an intergovernmental organisation that brings together life science resources from across Europe. To coordinate the linkage of various life science efforts around modern predictive toxicology, the establishment of a new ELIXIR Community is seen as instrumental. In the past few years, joint efforts, building on incidental overlap, have been piloted in the context of ELIXIR. For example, the EU-ToxRisk, diXa, HeCaToS, transQST, and the nanotoxicology community have worked with the ELIXIR TeSS, Bioschemas, and Compute Platforms and activities. In 2018, a core group of interested parties wrote a proposal, outlining a sketch of what this new ELIXIR Toxicology Community would look like. A recent workshop (held September 30th to October 1st, 2020) extended this into an ELIXIR Toxicology roadmap and a shortlist of limited investment-high gain collaborations to give body to this new community. This Whitepaper outlines the results of these efforts and defines our vision of the ELIXIR Toxicology Community and how it complements other ELIXIR activities.

The discovery of a drug requires over a decade of intensive research and financial investments – and still has a high risk of failure. To reduce this burden, we developed the NICEdrug.ch resource, which incorporates 250,000 bioactive molecules, and studied their enzymatic metabolic targets, fate, and toxicity. NICEdrug.ch includes a unique fingerprint that identifies reactive similarities between drug–drug and drug–metabolite pairs. We validated the application, scope, and performance of NICEdrug.ch over similar methods in the field on golden standard datasets describing drugs and metabolites sharing reactivity, drug toxicities, and drug targets. We use NICEdrug.ch to evaluate inhibition and toxicity by the anticancer drug 5-fluorouracil, and suggest avenues to alleviate its side effects. We propose shikimate 3-phosphate for targeting liver-stage malaria with minimal impact on the human host cell. Finally, NICEdrug.ch suggests over 1300 candidate drugs and food molecules to target COVID-19 and explains their inhibitory mechanism for further experimental screening. The NICEdrug.ch database is accessible online to systematically identify the reactivity of small molecules and druggable enzymes with practical applications in lead discovery and drug repurposing.

To assess the risks associated with cyanobacterial blooms, the persistence and fate processes of cyanotoxins and other bioactive cyanobacterial metabolites need to be evaluated. Here, we investigated the reaction with photochemically produced singlet oxygen (¹O₂) for 30 cyanopeptides synthesized by Dolichospermum flos aquae, including 9 anabaenopeptins, 18 microcystins, 2 cyanopeptolins, and 1 cyclamide. All compounds were stable in UVA light alone but in the presence of a photosensitizer we observed compound-specific degradation. A strong pH effect on the decay was observed for 18 cyanopeptides that all contained tyrosine or structurally related moieties. We can attribute this effect to the reaction with ¹O₂ and triplet sensitizer that preferentially react with the deprotonated form of tyrosine moieties. The contribution of ¹O₂ to indirect phototransformation ranged from 12 to 39% and second-order rate constants for 9 tyrosine-containing cyanopeptides were assessed. Including the pH dependence of the reaction and system-independent second-order rate constants with ¹O₂ will improve the estimation of half-lives for multiclass cyanopeptide in surface waters. Our data further indicates that naturally occurring triplet sensitizers are likely to oxidize deprotonated tyrosine moieties of cyanopeptides and the specific reactivity and its pH dependence needs to be investigated in future studies.

Background: Cyanobacteria and their toxins occur in high concentrations during the so-called bloom events in surface waters. To be able to assess the risks associated with cyanobacterial blooms, we need to understand the persistence and fate processes of these toxins and other bioactive metabolites. In this study, we investigated the photochemical fate of 54 cyanopeptides extracted from two strains of Microcystis aeruginosa (PCC7806 and UV006), Planktothrix rubescens, and Dolichospermum flos aquae. We determined half-lives during sunlight exposure in lake water and inspected the effect of pH on transformation kinetics for 27 microcystins, 8 anabaenopeptins, 14 cyanopeptolins, 2 cyclamides, and 3 aeruginosins.
Results: For cyanopeptides from D. flos aquae and P. rubescens, we observed the highest removal of 28 and 26%, respectively, after 3-h sunlight exposure. Most cyanopeptides produced by the two M. aeruginosa strains were rather persistent with only up to 3% removal. The more reactive cyanopeptides contained amino acids known to undergo phototransformation, including methionine and tyrosine moieties or their derivatives. Photochemical half-lives of 14 tyrosine-containing cyanopeptides decreased by one order of magnitude from nearly persistent conditions at pH 7 (half-life > 70 h) to shorter half-lives at pH 10 (< 10 h).
Conclusions: More work is needed to distinguish the contribution of different photochemical reaction pathways including the contributions to the pH effect. To the best of our knowledge, this is the first assessment of transformation kinetics of such a wide range of cyanopeptides. The abundant and persistent cyanopeptides that have not been studied in detail yet should be prioritized for the evaluation of their ecosystem and human health risks and for their abatement during drinking water treatment.

Landwirtschaftliche Nutztiere emittieren in der Schweiz rund achtmal weniger Estrogene auf Böden als der Mensch in die Abwasserreinigungsanlagen (ARA). Für die weitere Verteilung dieser Steroidhormone in der Umwelt gilt: Böden wie auch ARA sind höchst effiziente Filter. Mit einer Reihe von Messkampagnen und Feldversuchen konnte gezeigt werden, dass nur ein geringer Teil der via Gülle in die Umwelt eingetragenen Estrogene in die Gewässer gelangte und dies kaum zu längerfristig kritischen Konzentrationen führte. Sofern ARA mit im Spiel waren, stammten Estrogene selbst bei einer hohen Nutztierdichte überwiegend aus der Anthroposphäre.

Natural estrogens are present in high concentrations in livestock slurries, which are often applied to agricultural fields in large quantities. As such, the export of slurry-derived natural estrogens from tile-drained fields is a potential source for estrogenic pollution in surface waters. Yet despite the abundance of tile-drained fields in Central Europe, export of natural estrogens from agricultural fields receiving livestock slurries is rarely studied in this region. In an effort to fill this knowledge gap, here we applied natural estrogens to Swiss experimental fields in the form of cattle slurry, pig slurry or dissolved in water, and quantified them in flow-proportionally collected drainage water over 18 months. After pig and cattle slurry applications, concentration maxima in drainage water of 73, 8, 37, and 60 ng L⁻¹ for 17α-estradiol (E2α), 17β-estradiol (E2β), estrone (E1), and estriol (E3), respectively, were observed shortly after rain events. The exported fractions of individual natural estrogens to tile drains were on average 0.26% (ranging from 0.08 to 0.41%) after cattle, and 0.18% (0.03–0.40%) after pig slurry applications. Such numbers were higher than expected from the leaching potential based on substance properties, and comparable to those of more mobile micropollutants previously studied on the same fields. Natural estrogens were mainly exported to tile drains through preferential flow. Exported fractions were lower when applied in aqueous solution than when applied in slurry, pointing to particle-facilitated transport to tile drains when applied in slurry. In Switzerland, the estimated 6.3, 0.6, 7.4, and 7.7 g of E2α, E2β, E1, and E3, respectively, exported through the tile-drained agricultural area per year contribute little to the total natural estrogen load received by surface waters. However, after slurry applications in highly drained catchments, natural estrogen emissions to tile drains can cause short term (i.e. less than 1 week) estrogenic exposure in nearby streams.

Advanced treatment is increasingly being applied to improve abatement of micropollutants in wastewater effluent and reduce their load to surface waters. In this study, non-target screening of high-resolution mass spectrometry (HRMS) data, collected at three Swiss wastewater treatment plants (WWTPs), was used to evaluate different advanced wastewater treatment setups, including (1) granular activated carbon (GAC) filtration alone, (2) pre-ozonation followed by GAC filtration, and (3) pre-ozonation followed by powdered activated carbon (PAC) dosed onto a sand filter. Samples were collected at each treatment step of the WWTP and analyzed with reverse-phase liquid chromatography coupled to HRMS. Each WWTP received a portion of industrial wastewater and a prioritization method was applied to select non-target features potentially resulting from industrial activities. Approximately 37,000 non-target features were found in the influents of the WWTPs. A number of non-target features (1207) were prioritized as likely of industrial origin and 54 were identified through database spectral matching. The fates of all detected non-target features were assessed through a novel automated trend assignment method. A trend was assigned to each non-target feature based on the normalized intensity profile for each sampling date. Results showed that 73±4% of influent non-target features and the majority of industrial features (89%) were well-removed (i.e., >80% intensity reduction) during biological treatment in all three WWTPs. Advanced treatment removed, on average, an additional 11% of influent non-target features, with no significant differences observed among the different advanced treatment settings. In contrast, when considering a subset of 66 known micropollutants, advanced treatment was necessary to adequately abate these compounds and higher abatement was observed in fresh GAC (7,000-8,000 bed volumes (BVs)) compared to older GAC (18,000-48,000 BVs) (80% vs 56% of micropollutants were well-removed, respectively). Approximately half of the features detected in the WWTP effluents were features newly formed during the various treatment steps. In ozonation, between 1108-3579 features were classified as potential non-target ozonation transformation products (OTPs). No difference could be observed for their removal in GAC filters at the BVs investigated (70% of OTPs were well-removed on average). Similar amounts (67%) was observed with PAC (7.7-13.6 mg/L) dosed onto a sand filter, demonstrating that a post-treatment with activated carbon is efficient for the removal of OTPs.

Pesticide contamination of agricultural streams has widely been analysed in regions of high intensity agriculture such as in Western Europe or North America. The situation of streams subject to low intensity agriculture relying on human and animal labour, as in parts of Romania, remains unknown. To close this gap, we determined concentrations of 244 pesticides and metabolites at 19 low-order streams, covering sites from low to high intensity agriculture in a region of Romania. Pesticides were sampled with two passive sampling methods (styrene-divinylbenzene (SDB) disks and polydimethylsiloxane (PDMS) sheets) during three rainfall events and at base flow. Using the toxic unit approach, we assessed the toxicity towards algae and invertebrates. Up to 50 pesticides were detected simultaneously, resulting in sum concentrations between 0.02 and 37 µg L^-1. Both, the sum concentration as well as the toxicities were in a similar range as in high intensity agricultural streams of Western Europe. Different proxies of agricultural intensity did not relate to in-stream pesticide toxicity, contradicting the assumption of previous studies. The toxicity towards invertebrates was positively related to large scale variables such as the catchment size and the agricultural land use in the upstream catchment and small scale variables including riparian plant height, whereas the toxicity to algae showed no relationship to any of the variables. Our results suggest that streams in low intensity agriculture, despite a minor reported use of agrochemicals, exhibit similar levels of pesticide pollution as in regions of high intensity agriculture.

Non-target analysis (NTA) employing high-resolution mass spectrometry is a commonly applied approach for the detection of novel chemicals of emerging concern in complex environmental samples. NTA typically results in large and information-rich datasets that require computer aided (ideally automated) strategies for their processing and interpretation. Such strategies do however raise the challenge of reproducibility between and within different processing workflows. An effective strategy to mitigate such problems is the implementation of inter-laboratory studies (ILS) with the aim to evaluate different workflows and agree on harmonized/standardized quality control procedures. Here we present the data generated during such an ILS. This study was organized through the Norman Network and included 21 participants from 11 countries. A set of samples based on the passive sampling of drinking water pre and post treatment was shipped to all the participating laboratories for analysis, using one pre-defined method and one locally (i.e. in-house) developed method. The data generated represents a valuable resource (i.e. benchmark) for future developments of algorithms and workflows for NTA experiments.

Surface runoff represents a major pathway for pesticide transport from agricultural areas to surface waters. The influence of artificial structures (e.g. roads, hedges, and ditches) on surface runoff connectivity has been shown in various studies. In Switzerland, so-called hydraulic shortcuts (e.g. inlet and maintenance shafts of road or field storm drainage systems) have been shown to influence surface runoff connectivity and related pesticide transport. Their occurrence and their influence on surface runoff and pesticide connectivity have, however, not been studied systematically.
To address that deficit, we randomly selected 20 study areas (average size of 3.5 km²) throughout the Swiss plateau, representing arable cropping systems. We assessed shortcut occurrence in these study areas using three mapping methods, namely field mapping, drainage plans, and high-resolution aerial images. Surface runoff connectivity in the study areas was analysed using a 2×2 m digital elevation model and a multiple-flow algorithm. Parameter uncertainty affecting this analysis was addressed by a Monte Carlo simulation. With our approach, agricultural areas were divided into areas that are either directly, indirectly (i.e. via hydraulic shortcuts), or not at all connected to surface waters. Finally, the results of this connectivity analysis were scaled up to the national level, using a regression model based on topographic descriptors, and were then compared to an existing national connectivity model.
Inlet shafts of the road storm drainage system were identified as the main shortcuts. On average, we found 0.84 inlet shafts and a total of 2.0 shafts per hectare of agricultural land. In the study catchments, between 43 % and 74 % of the agricultural area is connected to surface waters via hydraulic shortcuts. On the national level, this fraction is similar and lies between 47 % and 60 %. Considering our empirical observations led to shifts in estimated fractions of connected areas compared to the previous connectivity model. The differences were most pronounced in flat areas of river valleys.
These numbers suggest that transport through hydraulic shortcuts is an important pesticide flow path in a landscape where many engineered structures exist to drain excess water from fields and roads. However, this transport process is currently not considered in Swiss pesticide legislation and authorization. Therefore, current regulations may fall short in addressing the full extent of the pesticide problem. However, independent measurements of water flow and pesticide transport to quantify the contribution of shortcuts and validating the model results are lacking. Overall, the findings highlight the relevance of better understanding the connectivity between fields and receiving waters and the underlying factors and physical structures in the landscape.

Studying aquatic biotransformation of chemicals in laboratory experiments, i.e., OECD 308 and OECD 309 studies, is required by international regulatory frameworks to prevent the release of persistent chemicals into natural water bodies. Here, we aimed to address several previously described shortcomings of OECD 308/309 studies regarding their variable outcomes and questionable environmental relevance by broadly testing and characterizing a modified biotransformation test system in which an aerated water column covers a thin sediment layer. Compared to standard OECD 308/309 studies, the modified system showed little inter-replicate variability, improved observability of biotransformation, and consistency with first-order biotransformation kinetics for the majority of 43 test compounds, including pharmaceuticals, pesticides, and artificial sweeteners. To elucidate the factors underlying the decreased inter-replicate variability compared to OECD 309 outcomes, we used multidimensional flow cytometry data and a machine learning-based cell type assignment pipeline to study cell densities and cell type diversities in the sediment and water compartments. Our here presented data on cell type composition in both water and sediment allows, for the first time, to study the behavior of microbial test communities throughout different biotransformation simulation studies. We found that sediment-associated microbial communities were generally more stable throughout the experiments and exhibited higher cell type diversity than the water column-associated communities. Consistently, our data indicate that aquatic biotransformation of chemicals can be most robustly studied in test systems providing a sufficient amount of sediment-borne biomass. While these findings favor OECD 308-type systems over OECD 309-type systems to study biotransformation at the water–sediment interface, our results suggest that the former should be modified toward lower sediment–water ratios to improve observability and interpretability of biotransformation.

Pesticides are used globally in agriculture, and can have negative effects on human health and ecosystems, especially when not handled as intended. Still, an increasing number of smallholder farmers in low- and middle-income countries are using expensive pesticide products to increase their yield. Due to their low socio-economic status and educational level, smallholder farmers are particularly vulnerable to the negative impacts of yield losses, but also of those from pesticides. When confronted with pests, smallholder farmers develop a need for information and seek out appropriate sources. However, little is known about how smallholders go about this process, whether this process differs for organic and conventional pest management strategies and if farmers also focus on risks of pest management practices. Agro-input dealer are supposed to provide information on pesticide risks, but they often focus more on selling products than services. There is a knowledge gap in literature on how agro-input dealers give advice, what products they sell, whether they follow laws and recommendations on best practice, and whether their practices and shop organization prevent pesticide risk situations. This in turn leads to farmers not knowing about risks, or not considering them to be relevant. While farmers have been assessed regarding their pesticide knowledge, attitudes and practices, rarely these insights are compared across contexts, identifying differences and commonalities. This, together with a lack in pesticide training, results in farmers not always following good agricultural practices, thereby affecting their own health, their communities’ and ecosystems. These issues are not resolved in disciplinary silos, but only through cross-sectoral and participatory research and interventions. This idea of a transdisciplinary, border-crossing research project named 'Comparative appraisal of pesticide use in tropical settings: exposure pathways, health effects and institutional determinants' laid the foundation for this dissertation.

Background: Pesticides can have negative effects on human and environmental health, especially when not handled as intended. In many countries, agro-input dealers sell pesticides to smallholder farmers and are supposed to provide recommendations on application and handling. This study investigates the role of agro-input dealers in transmitting safety information from chemical manufacturers to smallholder farmers, assesses the safety of their shops, what products they sell, and how agro-input dealers abide by laws and recommendations on best practices for preventing pesticide risk situations.
Methods: Applying a mixed-methods approach, we studied agro-input dealers in Central and Western Uganda. Structured questionnaires were applied to understand agro-input dealers' knowledge, attitude and practices on pesticides (n = 402). Shop layout (n = 392) and sales interaction (n = 236) were assessed through observations. Actual behavior of agro-input dealers when selling pesticides was revealed through mystery shopping with local farmers buying pesticides (n = 94).
Results: While 97.0% of agro-input dealers considered advising customers their responsibility, only 26.6% of mystery shoppers received any advice from agro-input dealers when buying pesticides. 53.2% of products purchased were officially recommended. Sales interactions focused mainly on product choice and price. Agro-input dealers showed limited understanding of labels and active ingredients. Moreover, 25.0% of shops were selling repackaged products, while 10.5% sold unmarked or unlabeled products. 90.1% of shops were lacking safety equipment. Pesticides of World Health Organization toxicity class I and II were sold most frequently. Awareness of health effects seemed to be high, although agro-input dealers showed incomplete hygiene practices and were lacking infrastructure. One reason for these findings might be that only 55.7% of agro-input dealers held a certificate of competency on safe handling of pesticides and even fewer (5.7%) were able to provide a government-approved up-to-date license.
Conclusion: The combination of interviews, mystery shopping and observations proved to be useful, allowing the comparison of stated and actual behavior. While agro-input dealers want to sell pesticides and provide the corresponding risk advice, their customers might receive neither the appropriate product nor sufficient advice on proper handling. In light of the expected increase in pesticide use, affordable, accessible and repeated pesticide training and shop inspections are indispensable.
 

Tracking the occurrence of a broad variety of chemicals in the aquatic environment at a high temporal resolution over extended time periods is challenging. Here, we present a transportable platform integrating sample preparation, liquid chromatography-high-resolution mass spectrometry measurement, automated data processing, and remote control and surveillance. The system is capable of quantifying pollutants at 20 min intervals with high sensitivity (limit of quantification down to 10 ng/L) and operated stably on-site over prolonged periods (<3 ppm mass drift over two weeks, <0.3 min retention time drift over six weeks). Field campaigns under different conditions demonstrated unprecedented insights into pollutant dynamics. Time-resolved measurements in a small creek revealed concentration peaks with up to 10-fold exceedances of environmental quality standards. The quantification of illicit drugs and their metabolites in raw wastewater distinguished fractions of consumed and discarded cocaine. Frequency analyses and data mining showed clusters of diurnal, episodic, and possibly industry-related pollutants, including newly identified substances. With this enabling technology the potential for researching and managing chemicals in natural and technical environments can be significantly expanded beyond our current capabilities, for example, real-time control in process engineering and sewer operation (water management and environmental toxicology), industrial surveillance, and wastewater-based epidemiology.

Subsurface contamination with the explosive hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) at ordnance production and testing sites is a problem because of the persistence, mobility, and toxicity of RDX and the formation of toxic products under anoxic conditions. While the utility of compound-specific isotope analysis for inferring natural attenuation pathways from stable isotope ratios has been demonstrated, the stable isotope fractionation for RDX reduction by iron-bearing minerals remains unknown. Here, we evaluated N and C isotope fractionation of RDX during reduction by Fe(II) associated with Fe minerals and natural sediments and applied N isotope ratios to the assessment of mineral-catalyzed RDX reduction in a contaminant plume and in sediment columns treated by in situ chemical reduction. Laboratory studies revealed that RDX was reduced to nitroso compounds without denitration and the concomitant ring cleavage. Fe(II)/iron oxide mineral-catalyzed reactions exhibited N isotope enrichment factors, ^ϵ_N, between -6.3±0.3‰ and -8.2±0.2‰, corresponding to an apparent ¹⁵N kinetic isotope effect of 1.04-1.05. The observed variations of the δ¹⁵N of ∼15‰ in RDX from groundwater samples suggested an extent of reductive transformation of 85% at an ammunition plant. Conversely, we observed masking of N isotope fractionation after RDX reduction in laboratory flow-through systems, which was presumably due to limited accessibility to reactive Fe(II).

Multielement isotope fractionation studies to assess pollutant transformation are well-established for point-source pollution but are only emerging for diffuse pollution by micropollutants like pesticides. Specifically, chlorine isotope fractionation is hardly explored but promising, because many pesticides contain only few chlorine atoms so that "undiluted" position-specific Cl isotope effects can be expected in compound-average data. This study explored combined Cl, N, and C isotope fractionation to sensitively detect biotic and abiotic transformation of the widespread herbicides and groundwater contaminants acetochlor, metolachlor, and atrazine. For chloroacetanilides, abiotic hydrolysis pathways studied under acidic, neutral, and alkaline conditions as well as biodegradation in two soils resulted in pronounced Cl isotope fractionation (ε_Cl from -5.0 ± 2.3 to -6.5 ± 0.7‰). The characteristic dual C-Cl isotope fractionation patterns (Λ_C–Cl from 0.39 ± 0.15 to 0.67 ± 0.08) reveal that Cl isotope analysis provides a robust indicator of chloroacetanilide degradation. For atrazine, distinct Λ_C–Cl values were observed for abiotic hydrolysis (7.4 ± 1.9) compared to previous reports for biotic hydrolysis and oxidative dealkylation (1.7 ± 0.9 and 0.6 ± 0.1, respectively). The 3D isotope approach allowed differentiating transformations that would not be distinguishable based on C and N isotope data alone. This first data set on Cl isotope fractionation in chloroacetanilides, together with new data in atrazine degradation, highlights the potential of using compound-specific chlorine isotope analysis for studying in situ pesticide degradation.

Pesticides are applied at high rates around the globe to protect crops from pest infestation. As a consequence, a broad spectrum of pesticides is found in surface waters. Together with their transformation products (TP), they can elicit adverse effects on aquatic organisms. Threats due to pesticide contamination to aquatic organisms is an especially concerning issue in tropical regions in Low- and Middle-Income Countries. These countries represent the most intensively used agricultural areas of the world and heavy rainfalls are expected to favor the transport of pesticides from the fields into surface water. Additionally, owing to the low level of economic development, environmental monitoring, risk assessment and implementation of risk mitigation strategies receive little attention. One country illustrating this situation is Costa Rica. This thesis set out to 1) monitor agricultural-driven pesticide pollution in tropical Costa Rican streams in order to describe the occurrence, concentrations and distribution of pesticides and pesticide transformation products (PPTP); 2) assess the risks by these PPTP to aquatic biota; and 3) identify relevant pathways of PPTP-transport from the field into the streams as a basis to propose mitigation options to reduce pesticide inputs. [...]

Pestizide werden weltweit in hohen Mengen ausgebracht, um Nutzpflanzen vor Schädlingsbefall zu schützen. Infolgedessen gelangt ein breites Spektrum an Pestiziden in Oberflächengewässer, wo sie zusammen mit ihren Transformationsprodukten (TP) schädliche Auswirkungen auf Wasserorganismen ausüben können. Die Bedrohung aquatischer Organismen durch Pestizide ist besonders in tropischen Gebieten in Schwellen- und Entwicklungsländern ein besorgniserregendes Thema. Diese Länder stellen die am intensivsten genutzten landwirtschaftlichen Gebiete der Welt dar und es ist zu erwarten, dass dort starke Regenfälle den Transport von Pestiziden von den Feldern in die Oberflächengewässer begünstigen. Aufgrund des geringen wirtschaftlichen Entwicklungsniveaus wird zudem der Umweltüberwachung, Risikobewertung und Umsetzung von Risikominderungsstrategien wenig Aufmerksamkeit geschenkt. Ein Land, das diese Situation veranschaulicht, ist Costa Rica. Ziele dieser Arbeit waren daher, 1) die landwirtschaftlich bedingte Pestizid-verschmutzung in tropischen Bächen Costa Ricas zu erfassen, um das Vorkommen, die Konzentrationen und die Verteilung von Pestiziden und Pestizidtransformationsprodukten (PPTP) zu beschreiben; 2) die Risiken durch diese PPTP für aquatische Organismen zu bewerten; und 3) relevante Pfade des PPTP-Transports vom Feld in die Flüsse zu identifizieren, welche als Grundlage zur Risikominderung herangezogen werden können. [...]

The emergence and spread of antibiotic resistance is a major societal challenge and new antibiotics are needed to successfully fight bacterial infections. Because the release of antibiotics into wastewater and downstream environments is expected to contribute to the problem of antibiotic resistance, it would be beneficial to consider the environmental fate of antibiotics in the development of novel antibiotics. In this article, we discuss the possibility of designing peptide-based antibiotics that are stable during treatment (e.g. in human blood), but rapidly inactivated through hydrolysis by peptidases after their secretion into wastewater. In the first part, we review studies on the biotransformation of peptide-based antibiotics during biological wastewater treatment and on the specificity of dissolved extracellular peptidases derived from wastewater. In the second part, we present first results of our endeavour to identify peptide bonds that are stable in human blood plasma and susceptible to hydrolysis by the industrially produced peptidase Subtilisin A.

Synthetic Plant Protection Products (PPPs) are a key element for a large part of today's global food systems. However, the transport of PPPs and their transformation products (TPs) to water bodies has serious negative effects on aquatic ecosystems. Small streams in agricultural catchments may experience pronounced concentration peaks given the proximity to fields and poor dilution capacity. Traditional sampling approaches often prevent a comprehensive understanding of PPPs and TPs concentration patterns being limited by trade-offs between temporal resolution and duration of the observation period. These limitations result in a knowledge gap for accurate ecotoxicological risk assessment and the achievement of optimal monitoring strategies for risk mitigation. We present here high-frequency PPPs and TPs concentration time-series measured with the autonomous MS²Field platform that combines continuous sampling and on-site measurements with a high-resolution mass spectrometer, which allows for overcoming temporal trade-offs. In a small agricultural catchment, we continuously measured 60 compounds at 20 minutes resolution for 41 days during the growing season. This observation period included 8 large and 15 small rain events and provided 2560 concentration values per compound.
To identify similarities and differences among the compound-specific concentration time-series, we analysed the entire dataset with positive matrix factorisation. Six factors sufficiently captured the overall complexity in concentration dynamics. While one factor reflected dilution during rainfall, five factors identified PPPs groups that seemed to share a common history of recent applications. The investigation per event of the concentration time-series revealed a surprising complexity of dynamic patterns; physico-chemical properties of the compounds did not influence the (dis)similarity of chemographs. Some PPPs concentration peaks led while others lagged by several hours the water level peaks during large events. During small events, water level peaks always preceded concentration peaks, which were generally only observed when the water levels had almost receded to pre-event levels. Thus, monitoring schemes relying on rainfall or water level as proxies for triggering sampling may lead to systematic biases.
The high temporal resolution revealed that the Swiss national monitoring integrating over 3.5 days underestimated critical concentration peaks by a factor of eight to more than 32, captured 3 out of 11 exceedances of legal acute quality standards (the relevant values in the Swiss Water Protection Law) and recorded 1 out of 9 exceedances of regulatory acceptable concentrations (the relevant values for the PPPs registration process).
MS²Field allowed for observing unexpected and overlooked pesticide dynamics with consequences for further research but also for monitoring. The large variability in timing of concentration peaks relative to water level calls for more in-depth analyses regarding the respective transport mechanisms. To perform these analyses, spatially distributed sampling and time-series of geo-referenced PPPs application data are needed.

The role of diffuse phosphorus (P) losses from agricultural land gained importance since technical and constructional measures in urban areas and industry have reached their potential in temperate regions. Phytoremediation strategies are a new suggestion to effectively reduce soil legacy P but until now, there is a lack of estimates on the time such strategies should take. With a rainfall-runoff model, spatial information on the hydrological risk of the catchment of Lake Baldegg (Switzerland) was generated and combined with a soil test P (CO₂-saturated water extraction) map. Based on these results, two potential soil target test P values (2.0 mg P (kg soil)^-1 target-P 1 and 1.6 mg P (kg soil)^-1 target-P 2) were set. A simple nonlinear mixed effects model was used to compare different balance and cropping scenarios to decrease soil test P. The relationship between P-balance (input - output) and soil test P was found to be exponential. The confidence interval for the predicted time necessary to reach target-P 2 after a cessation of P-fertilization on intensively managed grassland was 2 to 9 years depending on the initial soil test P. If fertilization is completely ceased, the predicted P-decline times were longer (8 to 32 years). The decline-time for the balance that is recommended for farmers in the catchment of a P-fertilization that covers 80% of the demand was predicted to be 11 to 47 years. The study emphasizes that P phytoextraction can be an effective and time and resource efficient mitigation strategy for catchments with high legacy P.

Antibiotics in the environment cause widespread concern as a result of their potent inhibitory action on microbial growth and their role in potentially creating selective conditions for proliferation of antibiotic resistant bacteria. Comprising a carbon skeleton, antibiotics should be amenable to microbial biodegradation, but this is still largely uncharted territory because of their simultaneous strong toxicity. In this study, we estimated potential antibiotics degradation by and effects on mixed microbial communities at concentrations sufficiently high to allow sensitive detection of biomass growth, but simultaneously, low enough to mitigate their toxic action. We used three different mixed inoculum sources freshly derived from freshwater, activated sludge or soil, and tested a series of 15 antibiotics from different classes at 1 mg C-carbon l⁻¹ dosage. Consistent community growth was observed for freshwater and activated sludge with ampicillin, erythromycin and chloramphenicol, and with sulfomethoxazole for activated sludge, which was accompanied by parent compound disappearance. Community growth could be attributed to a few subclasses of recognized cell types by using supervised machine-learning-based classifiers. Most other tested antibiotics resulted in inhibition of community growth on background assimilable organic carbon, concomitant with altered composition of the resulting communities. We conclude that growth-linked biodegradation of antibiotics at low concentrations may be present among typical environmental microbiota, but for a selected subset only, whereas for the majority of antibiotics negative effects prevail without any sign of productive growth.

Urine has great potential to be an effective fertilizer due to its high nutrient content, however, it can contain potentially worrying pharmaceuticals. Our objective was to study whether urine storage and aerobic biological treatment, i.e. nitrification, was sufficient to remove pharmaceuticals or an additional treatment with activated carbon was necessary to produce a fertilizer from urine. We investigated the abatement of twelve pharmaceuticals, including antibiotics and antivirals, in laboratory experiments representing the treatment steps of anaerobic storage of source-separated human urine, stabilization using partial and full nitrification under acclimatized and non-acclimatized conditions, and treatment of nitrified urine using powdered activated carbon (PAC). Two-month-long-term storage of urine was insufficient to substantially degrade the pharmaceuticals, except for hydrochlorothiazide (>90%). In the partial and full nitrification fed-batch reactors, atazanavir, ritonavir, and clarithromycin were rapidly removed, with biotransformation rate constants greater than 10 L g_SS⁻¹d⁻¹. Darunavir, emtricitabine, trimethoprim, N4-acetylsulfamethoxazole, sulfamethoxazole, atenolol, diclofenac, and hydrochlorothiazide were degraded slowly, with biotransformation rate constants of < 1 L g_SS⁻¹d⁻¹. With 200 mg PAC L⁻¹, at least 90% of each investigated pharmaceutical was removed. Yeast estrogen screen tests and bioluminescence inhibition tests revealed efficient removal of estrogenicity (99%) and toxicity (56%) using nitrification, and a reduction of 89% and 64%, respectively, using 200 mg PAC L⁻¹. With our study, we provide biotransformation rate constants of compounds never previously investigated. We also show that a combination of nitrification and PAC adsorption enables the production of a safe fertilizer with sufficiently low pharmaceutical concentrations and no removal of beneficial nutrients.

Biotransformation of chemical contaminants is of importance in various natural and engineered systems. However, in complex microbial communities and with chemical contaminants at low concentrations, our current understanding of biotransformation at the level of enzyme-chemical interactions is limited. Here, we explored an approach to identify associations between micropollutant biotransformation and specific gene products in complex microbial communities, using association mining between chemical and metatranscriptomic data obtained from experiments with activated sludge grown at different solid retention times. We successfully demonstrate proportional relationships between the measured rate constants and associated gene transcripts for nitrification as a major community function, but also for the biotransformation of two nitrile-containing micropollutants (bromoxynil and acetamiprid) and transcripts of nitrile hydratases, a class of enzymes that we experimentally confirmed to produce the detected amide transformation products. As these results suggest that metatranscriptomic information can indeed be quantitatively correlated with low abundant community functions such as micropollutant biotransformation in complex microbial communities, we proceeded to explore the potential of association mining to highlight enzymes likely involved in catalyzing less well-understood micropollutant biotransformation reactions. Specifically, we use the cases of nitrile hydration and oxidative biotransformation reactions to show that the consideration of additional experimental evidence (such as information on biotransformation pathways) increases the likelihood of detecting plausible novel enzyme-chemical relationships. Finally, we identify a cluster of mono- and dioxygenase fourth-level enzyme classes that most strongly correlate with oxidative micropollutant biotransformation reactions in activated sludge.

Stand-alone reverse osmosis (RO) has been proposed to produce high-quality drinking water from raw riverbank filtrate impacted by anthropogenic activities. To evaluate RO efficacy in removing organic micropollutants, biological analyses were combined with non-target screening using high-resolution mass spectrometry and open cheminformatics tools. The bank filtrate induced xenobiotic metabolism mediated by the aryl hydrocarbon receptor AhR, adaptive stress response mediated by the transcription factor Nrf2 and genotoxicity in the Ames-fluctuation test. These effects were absent in the RO permeate (product water), indicating the removal of bioactive micropollutants by RO membranes. In the water samples, 49 potentially toxic compounds were tentatively identified with the in silico fragmentation tool MetFrag using the US Environmental Protection Agency CompTox Chemicals Dashboard database. 5 compounds were confirmed with reference standards and 16 were tentatively identified with high confidence based on similarities to accurate mass spectra in open libraries. The bioactivity data of the confirmed chemicals indicated that 2,6-dichlorobenzamide and bentazone in water samples can contribute to the activation of AhR and oxidative stress response, respectively. The bioactivity data of 7 compounds tentatively identified with high confidence indicated that these structures can contribute to the induction of such effects. This study showed that riverbank filtration followed by RO could produce drinking water free of the investigated toxic effects.

Herbicide pollution in headwater streams due to agricultural practices is a major environmental concern and is characterized by episodic peak concentrations from fast transport paths. We rely on previous experimental studies in a small (1.2km²) agricultural catchment in the Swiss Plateau, to model dynamic diffuse herbicide pollution with emphasis on fast transport paths in a conceptual modelling framework at the catchment scale. We show how experimentalists’ understanding of the fate of herbicides (perceptual model) can be translated into conceptual models considering sorption, degradation, and fast transport of water and chemicals facilitated by impervious surfaces, tile drains and artificial shortcuts. Different types of experimental data (streamflow, high-frequency concentration measurements, and soil–water distribution coefficients) are used in a joint Bayesian inference of model parameters. We assess the ability of different spatial configurations of hydrological response units in explaining observed heterogeneity in transport behaviour of two corn herbicides. Thereby, we find that (1) relatively simple conceptual models can provide a realistic description of herbicide fate in small agricultural catchments, (2) accounting for spray drift onto hard surfaces is necessary to avoid a severe model bias during the first rainfall event after application, and (3) including catchment-specific experimentalist knowledge about important elements like artificial shortcuts and tile drains leads to a reduction in uncertainty of 30% compared to the more conventional conception of the proximity to the stream as the dominant risk factor.

This study presents a nontarget approach to detect discharges from pharmaceutical production in municipal wastewater treatment plant (WWTP) effluents and to estimate their relevance on the total emissions. Daily composite samples were collected for 3 months at two WWTPs in Switzerland, measured using liquid chromatography high-resolution mass spectrometry, and time series were generated for all features detected. The extent of intensity variation in the time series was used to differentiate relatively constant domestic inputs from highly fluctuating industrial emissions. We show that an intensity variation threshold of 10 correctly classifies compounds of known origin and reveals clear differences between the two WWTPs. At the WWTP receiving wastewater from a pharmaceutical manufacturing site, (i) 10 times as many potential industrial emissions were detected as compared to the WWTP receiving purely domestic wastewater; (ii) for 11 pharmaceuticals peak concentrations, >10 μg/L and up to 214 μg/L were quantified, which are clearly above typical municipal wastewater concentrations; and (iii) a pharmaceutical not authorized in Switzerland was identified. Signatures of potential industrial emissions were even traceable at the downstream Rhine monitoring station at a >4000-fold dilution. Several of them occurred repeatedly, suggesting that they were linked to regular production, not to accidents. Our results demonstrate that small wastewater volumes from a single industry not only left a clear signature in the effluents of the respective WWTP but also influenced the water quality of one of Europe's most important river systems. Overall, these findings indicate that pharmaceutical production is a relevant emission source even in highly developed countries with a strong focus on water quality, such as Switzerland.

In this work, emissions of active pharmaceutical ingredients (APIs) from formulating pharmaceutical industries (FPIs) were investigated for the first time based on detailed production information and compared to overall API emissions in wastewater treatment plant (WWTP) effluents. At two municipal WWTPs, both receiving wastewater from several FPIs, two months' daily effluent samples were collected and measured using liquid chromatography high-resolution mass spectrometry (LC-HRMS). Thirty-three APIs formulated during the sampling period as well as >120 organic contaminants commonly present in WWTP effluents were quantified. On the basis of their time patterns and manufacturing data, industrial contributions were found for 22 of 26 APIs (85%) detected in the samples and processed by the FPIs. API emissions from FPIs led to daily concentration increases of up to 300-fold, despite pretreatment of the industrial wastewater. However, emissions from FPIs seemed to depend on the type of formulating activity, with granulation and mixing being most prone to API losses. Losses from FPIs were responsible for the highest concentrations and for up to 60% of the daily total API emissions measured. Furthermore, screening for suspects in LC-HRMS data resulted in the detection of unexpected emissions from FPIs, demonstrating the value of these data to comprehensively assess industrial API losses. Overall, this study showed that FPIs were relevant contributors of APIs emitted in the WWTP effluents, although only a minor fraction (<1%) of the total processed API quantity was lost to the wastewater, and despite the small percentage (<5%) of FPI wastewater compared to the total wastewater flow.

Although the exposure assessment of wastewater-derived micropollutants via chemical, bioanalytical, and modeling methods in environmental compartments is becoming more frequent, the whole-body burden (i.e., internal concentrations) in nontarget organisms is rarely assessed. An understanding of the internal concentration fluctuation is especially important when exploring the mechanistic linkage between exposure and effects. In this study, we coupled a simple river model with a first-order toxicokinetic (TK) model to predict the concentrations of wastewater-derived micropollutants in freshwater invertebrates (Gammarus spp.). We applied Monte Carlo simulations and conducted laboratory experiments to account for the uncertain input data and the lack of uptake/depuration rate constants required for the TK model. The internal concentrations in field gammarids were predicted well, and the estimates varied only by a factor of 0.1-1.9. Fast equilibrium may also be assumed such that bioconcentration factors (BCFs) are used together with the daily river dilution patterns to predict internal concentrations. While this assumption is suitable for compounds observed in our experiment to reach the steady state within 48 h in gammarids, the model overpredicted the concentrations of substances that reach this condition after longer periods. Nevertheless, this approach provides conservative estimates and simplifies the coupling of models as BCFs are slightly more accessible than the rate constants. However, if one is interested in a more detailed exposure information (e.g., peak concentration and the whole-body burden recovery after a spill), then the nonsteady-state formulation should be employed.

Exposure assessment of pesticides has substantially improved over time, with methods that now include a combination of advanced analytical techniques and fate/transport models to evaluate their spatiotemporal distribution. However, the current regulatory environmental risk assessment considers thresholds from laboratory studies completed under standardized conditions that do not reflect environmental dynamics. Using the General Unified Threshold model for Survival (GUTS) model framework, we predicted the impact of time-varying pesticide exposures on the survival of gammarids in a small agricultural stream. The LP₅₀ values were used as an additional metric for assessing risks (defined in GUTS as a multiplication factor applied to the concentration time series to induce 50% mortality by the end of exposure). Although real-case exposures to individual pesticides were predicted to produce little to no impact on survival, the LP₅₀ values indicate acute (LP₅₀ ≤ 100) and/or chronic (LP₅₀ ≤ 10) toxicities for azoxystrobin, chlorpyrifos, diazinon, and imidacloprid, while risk to propiconazole exposure was considered very low (LP₅₀ ≫ 100). Finally, the model was extended to reflect mixture toxicity via concentration addition. It predicted risks under acute and chronic exposures to organophosphates and neonicotinoids. Given that gammarids are simultaneously exposed to multiple chemicals and other stressors throughout their lifetime, a decline in survival probabilities due to chemical stress can likely influence their overall fitness. We recognize that some assumptions require validation, but our work included a level of realism that can assist risk managers when evaluating the cumulative consequences of chemical exposure.

Ferrous iron-bearing minerals are important reductants in the contaminated subsurface, but their availability for the reduction of anthropogenic pollutants is often limited by competition with other electron acceptors including microorganisms and poor accessibility to Fe(II) in complex hydrogeologic settings. The supply of external electron donors through in situ chemical reduction (ISCR) has been proposed as one remediation approach, but the quantification of pollutant transformation is complicated by the perturbations introduced to the subsurface by ISCR. Here, we evaluate the application of compound specific isotope analysis (CSIA) for monitoring the reduction of 2,4-dinitroanisole (DNAN), a component of insensitive munitions formulations, by mineral-bound Fe(II) generated through ISCR of subsurface material from two field sites. Electron balances from laboratory experiments in batch and column reactors showed that 3.6% to 11% of the total Fe in the sediments was available for the reduction of DNAN and its partially reduced intermediates after dithionite treatment. The extent of DNAN reduction was successfully quantified from its N isotope fractionation measured in the column effluent based on the derivation of a N isotope enrichment factor, ϵ_N, derived from a comprehensive series of isotope fractionation experiments with numerous Fe(II)-bearing minerals as well as dithionite-reduced subsurface materials. Our observations illustrate the utility of CSIA as a robust approach to evaluate the success of in situ remediation through abiotic contaminant reduction.

Enzymatic oxygenations initiate biodegradation processes of many organic soil and water contaminants. Even though many biochemical aspects of oxygenation reactions are well-known, quantifying rates of oxidative contaminant removal as well as the extent of oxygenation remains a major challenge. Because enzymes use different strategies to activate O₂, reactions leading to substrate oxygenation are not necessarily limiting the rate of contaminant removal. Moreover, oxygenases react along unproductive pathways without substrate metabolism leading to O₂ uncoupling. Here, we identify the critical features of the catalytic cycles of selected oxygenases that determine rates and extents of biodegradation. We focus most specifically on Rieske dioxygenases, a subfamily of mononuclear non-heme ferrous iron oxygenases, because of their ability to hydroxylate unactivated aromatic structures and thus initiate the transformation of the most persistent organic contaminants. We illustrate that the rate-determining steps in their catalytic cycles range from O₂ activation to substrate hydroxylation, depending on the extent of O-O cleavage that is required for generating the reactive Fe-oxygen species. The extent of O₂ uncoupling, on the other hand, is highly substrate-specific and potentially modulated by adaptive responses to oxidative stress. Understanding the kinetic mechanisms of oxygenases will be key to assess organic contaminant biotransformation quantitatively.

Multiple anthropogenic drivers are changing ecosystems globally, with a disproportionate and intensifying impact on freshwater habitats. A major impact of urbanization are inputs from wastewater treatment plants (WWTPs). Initially designed to reduce eutrophication and improve water quality, WWTPs increasingly release a multitude of micropollutants (MPs; i.e., synthetic chemicals) and microbes (including antibiotic‐resistant bacteria) to receiving environments. This pollution may have pervasive impacts on biodiversity and ecosystem services. Viewed through multiple lenses of macroecological and ecotoxicological theory, we combined field, flume, and laboratory experiments to determine the effects of wastewater (WW) on microbial communities and organic‐matter processing using a standardized decomposition assay. First, we conducted a mensurative experiment sampling 60 locations above and below WWTP discharges in 20 Swiss streams. Microbial respiration and decomposition rates were positively influenced by WW inputs via warming and nutrient enrichment, but with a notable exception: WW decreased the activation energy of decomposition, indicating a "slowing" of this fundamental ecosystem process in response to temperature. Second, next‐generation sequencing indicated that microbial community structure below WWTPs was altered, with significant compositional turnover, reduced richness, and evidence of negative MP influences. Third, a series of flume experiments confirmed that although diluted WW generally has positive influences on microbial‐mediated processes, the negative effects of MPs are "masked" by nutrient enrichment. Finally, transplant experiments suggested that WW‐borne microbes enhance decomposition rates. Taken together, our results affirm the multiple stressor paradigm by showing that different aspects of WW (warming, nutrients, microbes, and MPs) jointly influence ecosystem functioning in complex ways. Increased respiration rates below WWTPs potentially generate ecosystem "disservices" via greater carbon evasion from streams and rivers. However, toxic MP effects may fundamentally alter ecological scaling relationships, indicating the need for a rapprochement between ecotoxicological and macroecological perspectives.

L'objectif du système modulaire gradué (SMG) est le dévelopment de méthodes standardisées permettant d'analyser et d'apprécier l' état des cours d'eau en Suisse. Des modules consacrés aux domaines de la biologie, de l’écomorphologie, de l’hydrologie, de la chimie et de l’écotoxicologie des eaux existent ou sont en cours de développement. Il manque ainsi un module qui porte sur l’évaluation de la qualité des sédiments. Le projet « Module Sédiment » a démarré en janvier 2015 au Centre Ecotox en partenariat avec l'OFEV et l'Eawag. Il avait pour objectif d'harmoniser les approches d'évaluation de la qualité des sédiments pour l'appréciation de l'état des cours d'eau en proposant une méthode d’échantillonnage des sédiments et des critères de qualité écotoxicologique utilisable par tous les cantons. Dans cet article, nous voulons présenter les principes et les éléments de la stratégie proposée qui sera publiée en tant que rapport d’expert au cours de l’année 2020.

The reduction of environmental pollutants by Fe²⁺ bound to iron oxides is an important process that determines pollutant toxicities and mobilities. Recently, we showed that pollutant reduction rates depend on the thermodynamic driving force of the reaction in a linear free energy relationship that was a function of the solution pH value and the reduction potential, E_H, of the interfacial Fe³⁺/Fe²⁺ redox couple. In this work, we studied how carbonate affected the free energy relationship by examining the effect that carbonate has on nitrobenzene reduction rates by Fe²⁺ bound to goethite (α-FeOOH). Carbonate slowed nitrobenzene reduction rates by inducing goethite particle aggregation, as evidenced by surface charge and particle size measurements. We observed no evidence for carbonate affecting Fe³⁺/Fe²⁺ reduction potentials or the mechanism of nitrobenzene reduction. The linear free energy relationship accurately described the data collected in the presence of carbonate when we accounted for the effect it had on the reactive surface area of goethite. The findings from this work provide a framework for determining why common groundwater constituents affect the E_H-dependence of reaction rates involving oxide-bound Fe²⁺ as a reductant.

An approach to identifying persistent organic contaminants in the environment was developed and executed for Switzerland as an example of an industrialized country. First, samples were screened with an in-house list using liquid chromatography high-resolution mass spectrometry (LC-HRMS/MS) and gas chromatography tandem mass spectrometry (GC-MS/MS) in 13 samples from the Swiss National Soil Monitoring Network and three sediment cores of an urban and agricultural contaminated lake. To capture a broader range of organic contaminants, the analysis was extended with a suspect screening analysis by LC-HRMS/MS of >500 halogenated compounds obtained from a Swiss database that includes industrial and household chemicals identified, by means of fugacity modeling, as persistent substances in the selected matrices. In total, the confirmation of 96 compounds with an overlap of 34 in soil and sediment was achieved. The identified compounds consist generally of esters, tertiary amines, trifluoromethyls, organophosphates, azoles and aromatic azines, with azoles and triazines being the most common groups. Newly identified compounds include transformation products, pharmaceuticals such as the flukicide niclofolan, the antimicrobial cloflucarban, and the fungicide mandipropamid. The results indicate that agricultural and urban soils as well as sediments impacted by agriculture and wastewater treatment plants (WWTPs) are the most contaminated sites. The plausibility of this outcome confirms the combination of chemical inventory, modeling of partitioning and persistence, and HRMS-based screening as a successful approach to shed light on less frequently or not yet investigated environmental contaminants and emphasizes the need for more soil and sediment monitoring in the future.

Aquatic pesticide pollution from both agricultural and urban pest control is a concern in many parts of the world. Making an accurate assessment of pesticide exposure is the starting point to protecting aquatic ecosystems. This in turn requires the design of an effective monitoring program. Monitoring is also essential to evaluate the efficacy of mitigation measures aimed to curb pesticide pollution. However, empirical evidence for their efficacy can be confounded by additional influencing factors, most prominently variable weather conditions. This review summarizes the experiences gained from long-term (>5 years) pesticide monitoring studies for detecting trends and provides recommendations for their improvement. We reviewed articles published in the scientific literature, with a few complements from selected grey literature, for a total of 20 studies which fulfill our search criteria. Overall, temporal trends of pesticide use and hydrological conditions were the two most common factors influencing aquatic pesticide pollution. Eighteen studies demonstrated observable effects to surface water concentrations from changes in pesticide application rates (e.g., use restriction) and sixteen studies from interannual variability in hydrological conditions during the application period. Accounting for seasonal- and streamflow-related variability in trend analysis is important because the two factors can obscure trends caused by changes in pesticide use or management practices. Other mitigation measures (e.g., buffer strips) were only detectable in four studies where concentrations or loads were reduced by > 45%. Collecting additional agricultural (e.g., pesticide use, mitigation measures) and environmental (e.g., precipitation, stream flow) data, as well as establishing a baseline before the implementation of mitigation measures have been consistently reported as prerequisites to interpret water quality trends from long-term monitoring studies, but have rarely been implemented in the past.

Assessment of micropollutant biodegradation is essential to determine the persistence of potentially hazardous chemicals in aquatic ecosystems. We studied the dissipation half-lives of 10 micropollutants in sediment–water incubations (based on the OECD 308 standard) with sediment from two European rivers sampled upstream and downstream of wastewater treatment plant (WWTP) discharge. Dissipation half-lives (DT50s) were highly variable between the tested compounds, ranging from 1.5 to 772 days. Sediment from one river sampled downstream from the WWTP showed the fastest dissipation of all micropollutants after sediment RNA normalization. By characterizing sediment bacteria using 16S rRNA sequences, bacterial community composition of a sediment was associated with its capacity for dissipating micropollutants. Bacterial amplicon sequence variants of the genera Ralstonia, Pseudomonas, Hyphomicrobium, and Novosphingobium, which are known degraders of contaminants, were significantly more abundant in the sediment incubations where fast dissipation was observed. Our study illuminates the limitations of the OECD 308 standard to account for variation of dissipation rates of micropollutants due to differences in bacterial community composition. This limitation is problematic particularly for those compounds with DT50s close to regulatory persistence criteria. Thus, it is essential to consider bacterial community composition as a source of variability in regulatory biodegradation and persistence assessments.

Lipids play a significant role in regulation of health and disease. To enhance our understanding of the role of lipids in regulation of lifespan and healthspan additional studies are required. Here, UHPLC-MS/MS lipidomics was used to measure dynamic changes in lipid composition as a function of age and gender in genetically identical male and female Daphnia magna with different average lifespans. We demonstrate statistically significant age-related changes in triglycerides (TG), diglycerides (DG), phosphatidylcholine, phosphatidylethanolamine, ceramide and sphingomyelin lipid groups, for example, in males, 17.04% of TG lipid species decline with age whilst 37.86% increase in relative intensity with age. In females, 23.16% decrease and 25.31% increase in relative intensity with age. Most interestingly, the rate and direction of change can differ between genetically identical female and male Daphnia magna, which could be the cause and/or the consequence of the different average lifespans between the two genetically identical genders. This study provides a benchmark dataset to understand how lipids alter as a function of age in genetically identical female and male species with different average lifespan and ageing rate.

Environmental risk assessment associated with aquatic and terrestrial contamination is mostly based on predicted or measured environmental concentrations of a limited list of chemicals in a restricted number of environmental compartments. High resolution mass spectrometry (HRMS) can provide a more comprehensive picture of exposure to harmful chemicals, particularly through the retrospective analysis of digitally stored HRMS data. Using this methodology, our study characterized the contamination of various environmental compartments including 154 surface water, 46 urban effluent, 67 sediment, 15 soil, 34 groundwater, 24 biofilm, 41 gammarid and 49 fish samples at 95 sites widely distributed over the Swiss Plateau. As a proof-of-concept, we focused our investigation on antifungal azoles, a class of chemicals of emerging concern due to their endocrine disrupting effects on aquatic organisms and humans. Our results demonstrated the occurrence of antifungal azoles and some of their (bio)transformation products in all the analyzed compartments (0.1-100 ng/L or ng/g d.w.). Comparison of actual and predicted concentrations showed the partial suitability of level 1 fugacity modelling in predicting the exposure to azoles. Risk quotient calculations additionally revealed risk of exposure especially if some of the investigated rivers and streams are used for drinking water production. The case study clearly shows that the retrospective analysis of HRMS/MS data can improve the current knowledge on exposure and the related risks to chemicals of emerging concern and can be effectively employed in the future for such purposes.

South Africa is the leading pesticide user in Sub-Saharan Africa. However, little is known about the occurrence of pesticide mixtures in surface water and potential environmental risks in Africa.
This study investigated the occurrence of pesticides mixtures in three watersheds during a drought year in South Africa. The study was conducted in the Krom River, Berg River and Hex River watersheds within larger agriculture systems in the Western Cape. Pesticide spray records were collected from 38 farms. A total of 21 passive water samplers (styrenedivinylbenzene disks (SDB)) were deployed, each for two weeks per month, over seven sampling rounds during the main pesticide application period between July 2017 and January 2018. Samples were analyzed for 248 pesticide compounds using LC-HR-MS/MS. Pesticide occurrence was analyzed for temporal agreement with pesticide spraying events (Cohen’s κ) and correlation with rainfall patterns and river discharge (Pearson correlation (r_p)). Pesticide time-weighted average concentrations were estimated and compared to environmental quality standards (EQS). According to the farm spray records, 96 different pesticides were sprayed during the sampling period and differed considerably between the three study areas, seasons and crops grown. In total, 53 compounds were detected in river water. We detected 39% of compounds from the spraying records and demonstrated close temporal correlations of seasonal patterns for 11 pesticide compounds between reported on spraying records and observations in the streams (κ = 0.90). However, 23 detected pesticides were not found on spray records, many of them being herbicides. Most of the estimated two-week average pesticide concentrations were below 40 ng/L. The insecticides imidacloprid, thiacloprid, chlorpyrifos and acetamiprid and the herbicide terbuthylazine exceeded at least once their EQS 58-fold (EQS 13 ng/L), 12-fold (EQS 10 ng/L), 9-fold (EQS 0.46 ng/L), 5-fold (EQS 24 ng/L) and 3-fold (EQS 220 ng/L), respectively. Our study substantially widens the view on pesticide pollution in surface water compared to previous studies in Sub-Saharan Africa by targeting more than 200 pesticides using passive sampling systems. This broad assessment revealed the presence of 53 compounds, some of them in high concentrations, indicating possible adverse effects on biota and the quality of the ecosystem. Whether the observed concentration levels in the year 2017 were exceptional due to the lowest ever recorded rainfall and river discharge needs to be tested with additional data to better understand how pesticide pollution levels manifest under average rainfall and river discharge conditions.

Antimicrobial peptides (AMPs) are increasingly important as a last resort against multi-drug resistant bacteria due to resistance formation towards conventional antibiotics. However, many AMPs were introduced to the market before environmental risk assessment was required, e.g., by the European Medicines Agency (EMA) since 1998. While AMPs have been administered as antibiotics and growth promotors in feedstock since the 1960s and were reconsidered for human medicine by the EMA in 2013, details about their mobility and persistence in the environment remain unknown. This study investigated the environmental fate of three commonly used AMPs: bacitracins, daptomycin, and polymyxins B and E (Colistin). We observed moderate sorption affinity of daptomycin to standard European soils (K_d = 20.6–48.6), while polymyxins adsorbed irreversibly. Bacitracin variants sorbed slightly to sandy soil (K_d = 5.8–8) and significantly to clayey soil (K_d = 169–250). We further investigated photochemical and microbial transformation processes relevant in surface waters. We demonstrated that phototransformation of all AMPs was enhanced in the presence of dissolved organic matter and fast bimolecular reaction rate constant with singlet oxygen contributed largely to indirect phototransformation (15–41%). Phototransformation product analysis for daptomycin was consistent with expected modifications of the tryptophan and kynurenine moieties. Moreover, riverine biofilm communities demonstrated biotransformation potential for all AMPs. Our findings of sorption behaviour, photo- and biotransformation suggest that these processes play a critical role in the fate of bacitracins, daptomycin, and polymyxins in environmental systems.

Konzentrationsspitzen von Pestiziden können von grosser biologischer und regulatorischer Bedeutung sein, werden aber selten und nur mit hohem Aufwand erfasst. Die neue MS²field-Plattform ist eine der ersten mobilen Messinfrastrukturen, die kontinuierliche und zeitlich hochaufgelöste Messungen von Mikroverunreinigungen in umweltrelevanten Konzentrationen im Feld erlaubt. Wie vielfältig Konzentrationsdynamiken von Pestiziden sein können und in welchem Ausmass Spitzenkonzentrationen mit konventionellen Probenahme- und Messtechniken bisher unterschätzt wurden, macht eine Messkampagne in einem landwirtschaftlich stark geprägten Bach sichtbar.

Conventional pesticides are associated with numerous human and environmental health risks. Nevertheless, an increasing number of smallholder farmers in low- and middle-income countries are using conventional pesticides. Adopting safer pest management requires farmers to obtain new information. However, little is known how farmers develop an information need, seek, and use pest management related information, and whether this process differs for organic and conventional pest management strategies. In this qualitative study, we investigated pest-related information behavior in depth, from farmers' own perspective. Using an ethnographic approach, we conducted 46 semi-structured interviews, 15 on-farm observations and 302 structured questionnaire interviews with farmers in Wakiso District, Uganda, in 2017. Our results indicated that farmers develop information needs when adopting new farming practices, or when presented with disruptive information (e.g. when new pests emerged). This prompted farmers to seek information actively, or they received passive information. Whether farmers used the new information depended on successful trial of the new pest management strategy, and on the credibility of the source. Most revealing, our results suggested important differences in information behavior between conventional and organic pest management strategies. Sources of information for conventional pesticides were well-integrated into farmers' daily lives and comprised pesticide dealers and fellow farmers. Conversely, information on organic strategies was provided through external sources (e.g. NGOs), and was not available at times when farmers developed an information need. Our results imply that farmers are most likely receptive to organic pest management information at times when they develop an information need (e.g. when encountering a new pest). To promote safer pest management, information about organic and integrated pest management should be made continuously available in farmers' lives. Furthermore, we recommend leveraging established information channels (e.g. dealers) among pesticide users to promote safer use practices.

Gemäss Verfassungsauftrag ist die Schweizer Landwirtschaft stark multifunktional ausgerichtet. Diese Multifunktionalität kann Zielkonflikte auslösen zwischen der sicheren Versorgung der Bevölkerung, der Erhaltung der natürlichen Lebensgrundlagen, der Pflege der Kulturlandschaft und der dezentralen Besiedlung des Landes. Um derartige Zielkonflikte systematisch zu analysieren, künftige Forschungsvorhaben zu priorisieren und Massnahmen zur Ressourcenschonung zu formulieren, haben die Eawag, das Wasserforschungsinstitut des ETH-Bereichs, und das BLW unter Einbezug von Agroscope 2013 bis 2015 das Projekt Zukunftsfähige, gewässerschonende landwirtschaftliche Produktion in der Schweiz (AProWa) durchgeführt. Methodisch wurde die umfassende Auslegeordnung in eine multikriterielle Entscheidungsanalyse (MCDA) eingebettet. Folgende Resultate wurden durch das Projekt erarbeitet:
1. Eine systematische Aufstellung und Gliederung von Produktionszielen (z. B. hohe Ertragsmenge) und Erhaltungszielen (z. B. geringe Schadstoffkonzentrationen im Grundwasser) in Form einer Zielhierarchie.
2. Ein Vorschlag an messbaren Indikatoren («Attribute» genannt) zur Quantifizierung der Zielerreichung.
3. Eine Sammlung von relevanten Massnahmen im Bereich einer gewässerschonenden Landwirtschaft.
4. Testen des MCDA-Ansatzes zur quantitativen Bewertung verschiedener Handlungsoptionen hinsichtlich ihrer Wirkungen im Gesamtsystem.
5. Vorschläge für zukünftige Forschungsfragen basierend auf dem aufgezeigten Forschungsbedarf.
6. Vorschläge für den weiteren Einsatz der MCDA-Methode in der Praxis. [ ...]

The Partnership for Chemicals Risk Assessment (PARC) is currently under development as a joint research and innovation programme to strengthen the scientific basis for chemical risk assessment in the EU. The plan is to bring chemical risk assessors and managers together with scientists to accelerate method development and the production of necessary data and knowledge, and to facilitate the transition to next-generation evidence-based risk assessment, a non-toxic environment and the European Green Deal. The NORMAN Network is an independent, well-established and competent network of more than 80 organisations in the field of emerging substances and has enormous potential to contribute to the implementation of the PARC partnership. NORMAN stands ready to provide expert advice to PARC, drawing on its long experience in the development, harmonisation and testing of advanced tools in relation to chemicals of emerging concern and in support of a European Early Warning System to unravel the risks of contaminants of emerging concern (CECs) and close the gap between research and innovation and regulatory processes. In this commentary we highlight the tools developed by NORMAN that we consider most relevant to supporting the PARC initiative: (i) joint data space and cutting-edge research tools for risk assessment of contaminants of emerging concern; (ii) collaborative European framework to improve data quality and comparability; (iii) advanced data analysis tools for a European early warning system and (iv) support to national and European chemical risk assessment thanks to harnessing, combining and sharing evidence and expertise on CECs. By combining the extensive knowledge and experience of the NORMAN network with the financial and policy-related strengths of the PARC initiative, a large step towards the goal of a non-toxic environment can be taken.

Extracellular enzymes are master recyclers of organic matter, and to predict their functional lifetime, we need to understand their environmental transformation processes. In surface waters, direct and indirect photochemical transformation is a known driver of inactivation. We investigated molecular changes that occur along with inactivation in aminopeptidase, an abundant class of extracellular enzymes. We studied the inactivation kinetics and localized oxidation caused by singlet oxygen, ¹O₂, a major photochemically derived oxidant toward amino acids. Aminopeptidase showed second-order inactivation rate constants with ¹O₂ comparable to those of free amino acids. We then visualized site-specific oxidation kinetics within the three-dimensional protein and demonstrated that fastest oxidation occurred around the active site and at other reactive amino acids. However, second-order oxidation rate constants did not correlate strictly with the ¹O₂-accessible surface areas of those amino acids. We inspected site-specific processes by a comprehensive suspect screening for 723,288 possible transformation products. We concluded that histidine involved in zinc coordination at the active site reacted slower than what was expected by its accessibility, and we differentiated between two competing reaction pathways of ¹O₂ with tryptophan residues. This systematic analysis can be directly applied to other proteins and transformation reactions.

Harmful cyanobacterial blooms in freshwater ecosystems produce bioactive secondary metabolites including cyanopeptides that pose ecological and human health risks. Only adverse effects of one class of cyanopeptides, microcystins, have been studied extensively and have consequently been included in water quality assessments. Inhibition is a commonly observed effect for enzymes exposed to cyanopeptides and has mostly been investigated for human biologically relevant model enzymes. Here, we investigated the inhibition of ubiquitous aquatic enzymes by cyanobacterial metabolites. Hydrolytic enzymes are utilized in the metabolism of aquatic organisms and extracellularly by heterotrophic bacteria to obtain assimilable substrates. The ubiquitous occurrence of hydrolytic enzymes leads to the co-occurrence with cyanopeptides especially during cyanobacterial blooms. Bacterial leucine aminopeptidase and alkaline phosphatase were exposed to cyanopeptide extracts of different cyanobacterial strains (Microcystis aeruginosa wild type and microcystin-free mutant, Planktothrix rubescens) and purified cyanopeptides. We observed inhibition of aminopeptidase and phosphatase upon exposure, especially to the apolar fractions of the cyanobacterial extracts. Exposure to the dominant cyanopeptides in these extracts confirmed that purified microcystins, aerucyclamide A and cyanopeptolin A inhibit the aminopeptidase in the low mg L^-1 range while the phosphatase was less affected. Inhibition of aquatic enzymes can reduce the turnover of nutrients and carbon substrates and may also impair metabolic functions of grazing organisms.

Compartment-specific degradation half-lives are essential pieces of information in the regulatory risk assessment of synthetic chemicals. However, their measurement according to regulatory testing guidelines is laborious and costly. Despite the obvious ecological and economic benefits of knowing environmental degradability as early as possible, its consideration in the early phases of rational chemical design is therefore challenging. Here, we explore the possibility to use half-lives determined in highly time- and work-efficient biotransformation experiments with activated sludge and mixtures of chemicals to predict soil half-lives from regulatory simulation studies. We experimentally determined half-lives for 52 structurally diverse agrochemical active ingredients in batch reactors with three concentrations of the same activated sludge. We then developed bi- and multivariate models for predicting half-lives in soil by regressing the experimentally determined half-lives in activated sludge against average soil half-lives of the same chemicals extracted from regulatory data. The models differed in how we accounted for sorption-related bioavailability differences in soil and activated sludge. The best-performing models exhibited good coefficients of determination (R² of around 0.8) and low average errors (

Biotransformation plays a crucial role in regulating the bioaccumulation potential and toxicity of organic compounds in organisms but is, in general, poorly understood for emerging contaminants. Here we have used diclofenac as a model compound to study the impact of biotransformation on the bioaccumulation potential and toxicity in two keystone aquatic invertebrates: Gammarus pulex and Hyalella azteca. In both species, diclofenac was transformed into several oxidation products and conjugates, including two novel products, i.e. diclofenac taurine conjugate (DCF-M403) and unexpected diclofenac methyl ester (DCF-M310.03). The ratios of biotransformation products to parent compound were 12-17 for DCF-M403 and 0.01-0.7 for DCF-M310.03 after 24 h exposure. Bioconcentration factors (BCFs) of diclofenac were 0.5 and 3.2 L kg ww^-1 in H. azteca and G. pulex, respectively, whereas BCFs of DCF-M310.03 was 164.5 L kg ww^-1 and 104.7 L kg ww^-1, respectively, representing a 25 to 110-fold increase. Acute toxicity of DCF-M310.03 was also higher than the parent compound in both species, which correlated well with the increased bioconcentration potential. The LC50 of diclofenac in H. azteca was 216 mg L^-1, while that of metabolite DCF-M310.03 was reduced to only 0.53 mg L^-1, representing a 430-fold increase in acute toxicity compared to diclofenac. DCF-M403 is less toxic than its parent compound towards H. azteca, which may be linked to its slightly lower hydrophobicity. Furthermore, the transformation of diclofenac to its methyl ester derivative was explored in crude invertebrate extracts spiked with an S-adenosylmethionine cofactor, revealing possible catalysis by an S-adenosylmethionine-dependent -carboxylic acid methyltransferase. Methylation of diclofenac was further detected in fish hepatocytes and human urine, indicating a broader relevance. Therefore, potentially methylated metabolites of polar contaminants should be considered for a comprehensive risk assessment in the future.

Objective. Estimates of pesticide exposure among applicators from low- and middle-income countries (LMICs) are scarce, and exposure assessment methods are sometimes costly or logistically unfeasible. We examined the variability in weeklong pesticide exposure among applicators in Costa Rica and its predictors.
Methods. We conducted a cross-sectional survey among 221 pesticide applicators from organic, sustainable and conventional farms in 2016. We administered structured questionnaires to assess pesticide application practices at two time points (4–6 weeks apart). We adapted an existing algorithm to fit the context of smallholders and derive weekly pesticide exposure scores. We used linear mixed-effect models to examine within-worker and between-worker score variability. We then identified sociodemographic and occupational predictors of weekly pesticide exposure scores.
Results. We observed high within-worker and between-worker variability in weekly pesticide exposures (eg, up to 180-fold and 70-fold differences in average weekly exposures within and between workers, respectively; intraclass correlation coefficient=0.4). Applicators working on conventional and sustainable farms had exposure scores twofold and 1.5-fold higher than those working in organic farms, respectively. Farm workers who received training on pesticide use had weekly pesticide exposure scores of 33% (95% CI 1% to 55%) lower than those who did not receive any training.
Conclusions. In this study of applicators from smallholder farms in Costa Rica, we determined the importance of collecting questionnaire data on self-reported pesticide use repeatedly due to its high variability within workers and absence of application records. Our questionnaire-based exposure algorithm could allow the calculation of semiquantitative estimates of average pesticide exposure for applicators from other LMICs.

This study presents the development and validation of a comprehensive quantitative target methodology for the analysis of 2316 emerging pollutants in water based on Ultra-Performance Liquid Chromatography Quadrupole-Time-Of-Flight Mass Spectrometry (UPLC-Q-ToF-HRMS/MS). Target compounds include pesticides, pharmaceuticals, drugs of abuse, industrial chemicals, doping compounds, surfactants and transformation products, among others. The method was validated for 195 analytes, chosen to be representative of the chemical space of the target list, enabling the assessment of the performance of the method. The method involves a generic sample preparation based on mixed mode solid phase extraction, a UPLC-QTOF-MS/MS screening method using Data Independent Acquisition (DIA) mode, which provides MS and MS/MS spectra simultaneously and an elaborate strong post-acquisition evaluation of the data. The processing method was optimized to provide a successful identification rate >95 % and to minimize the number of false positive results (< 5 %). Decision limit (CCα) and detection capability (CCβ) were also introduced in the validation scheme to provide more realistic metrics on the performance of a HRMS-based wide-scope screening method. A new system of identification points (IPs) based on the one described in the Commission Decision 2002/657/EC was applied to communicate the confidence level in the identification of the analytes. This system considers retention time, mass accuracy, isotopic fit and fragmentation; taking full advantage of the capacities of the HRMS instruments. Finally, 398 contaminants were detected and quantified in real wastewater.

Sertraline is one of the most commonly prescribed antidepressants in the last few years. Therefore, it is not surprising that it is regularly detected in wastewaters, surface waters, sediments, biosolids and biota. Effluents from wastewater treatment plants are the main contributors to its presence in the environment. The presented study aims to elucidate the processes involved in its removal, concentrating mainly on sorption and biodegradation during wastewater treatment.
We performed our laboratory scale experiments in two sets of experiments: 1) batch biodegradation and sorption experiments and 2) flow-through laboratory scale pilot wastewater treatment bioreactors. The batch experiments revealed that sorption to activated sludge was the leading removal process, eliminating up to 90% of sertraline present in the batches. Biodegradation was however the secondary removal process, influenced by the presence of alternative easily biodegradable carbon sources. We postulated chemical structures of ten detected biotransformation products. Among these, we propose the previously recognized metabolite norsertraline, sertraline ketone and hydroxy-sertraline. All the remaining biotransformation products are herein reported for the first time.
The removal efficiency of approximately 94% was determined after the treatment in the flow-through bioreactors. To support our findings, we sampled influents and effluents from two wastewater treatment plants and untreated wastewater from a psychiatric hospital. Removal efficiencies of 81% and 77% were determined, and along with the parent compound sertraline, the presence of eight transformation products was confirmed in the actual wastewaters.
 

In der Schweiz gelangt nach wie vor zu viel Stickstoff und Phosphor in die Umwelt. Kritische Belastungsgrenzen für Stickstoffeinträge in die Umwelt sind vielerorts deutlich überschritten. Vor allem die Stickstoff- und Phosphorüberschüsse der Landwirtschaft und die Stickstoffemissionen des Verkehrs belasten die Umwelt und beeinträchtigen die Biodiversität, die Luft, die Gewässer- und Trinkwasserqualität sowie die Waldfunktionen stark. Zudem verstärken sie den Klimawandel und beeinträchtigen die menschliche Gesundheit. Ursachen wie auch Auswirkungen der übermässigen Einträge sind wissenschaftlich seit Jahrzehnten gut dokumentiert. Allerdings ist kaum eines der diesbezüglichen vom Bundesrat verabschiedeten Reduktionsziele erreicht. Sollen die negativen Effekte auf die Biodiversität und Ökosystemleistungen verringert werden, sind die Ursachen für die übermässigen Einträge dringend anzugehen. Dieses Faktenblatt zeigt hierzu Handlungsansätze auf.

To protect themselves, plants can produce toxic secondary metabolites (phytotoxins) that appear with widely varying structures and negative effects. These phytotoxins often show similar properties as known aquatic micropollutants in terms of mobility, persistence, toxicity, and possibly also ecotoxicity. However, their occurrence in surface waters remains largely unknown, which is also due to unknown ability of available screening approaches to detect them. Therefore, we performed a target and suspect screening based on a persistence-mobility prioritization for phytotoxins in small Swiss creeks using high resolution mass spectrometry. In total, three of 26 targets were detected, three of 78 suspects tentatively identified, and six suspects fully confirmed by reference standards. To the best of our knowledge, it is the first time that three different plant secondary metabolite classes are detected in the same surface water sample. Estrogenic isoflavones were detected at 73% of the sites with formononetin as main toxin, which is in agreement with previous studies. Furthermore, pyrrolizidine alkaloids and the indole alkaloid gramine were detected. Especially pyrrolizidine alkaloids might be critical due to their production by various plants including the invasive Senecio inaequidens, and their known importance in food and feed safety. Based on these first screening results, different phytotoxin classes should be assessed for their ecotoxicological effects and considered in future water monitoring.

Implementing advanced wastewater treatment (WWT) to eliminate organic micropollutants (OMPs) is a necessary step to protect vulnerable freshwater ecosystems and water resources. To this end, sorption of OMP by activated carbon (AC) is one viable technology among others. However, conventional AC production based on fossil precursor materials causes environmental pollution, including considerable emissions of greenhouse gases. In this study, we produced activated biochar (AB) from wood and woody residues by physical activation and evaluated their capability to eliminate OMPs in treated wastewater. Activated biochar produced under optimized conditions sorbed 15 model OMPs, of which most were dissociated at circumneutral pH, to the same or higher extent than commercial AC used as a reference. While wood quality played a minor role, the dosage of the activation agent was the main parameter controlling the capacity of ABs to eliminate OMP. Our results highlight the possibility for local production of AB from local wood or woody residues as a strategy to improve WWT avoiding negative side effects of conventional AC production.

Objectives The risk of diabetes mellitus may be elevated among persons exposed to some pesticides, including cholinesterase-inhibiting insecticides (organophosphates and carbamates). The objective of this study was to investigate how acetylcholinesterase activity was associated with mean blood glucose levels among smallholder farmers in Uganda.
Methods We conducted a short-term follow-up study among 364 smallholder farmers in Uganda. Participants were examined three times from September 2018 to February 2019. At each visit, we measured glycosylated haemoglobin A (HbA_1c) as a measure of long-term average blood glucose levels. Exposure to organophosphate and carbamate insecticides was quantified using erythrocyte acetylcholinesterase normalised by haemoglobin (AChE/Hb). For a subgroup of participants, fasting plasma glucose (FPG) was also available. We analysed HbA_1c and FPG versus AChE/Hb in linear mixed and fixed effect models adjusting for age, sex, physical activity level, and consumption of fruits and vegetables, alcohol and tobacco.
Results Contrary to our hypothesis, our mixed effect models showed significant correlation between low AChE/Hb and low HbA_1c. Adjusted mean HbA_1c was 0.74 (95% CI 0.17 to 1.31) mmol/mol lower for subjects with AChE/Hb=24.3 U/g (35th percentile) compared with subjects with AChE/Hb=25.8 U/g (50th percentile). Similar results were demonstrated for FPG. Fixed effect models showed less clear correlations for between-phase changes in AChE/Hb and HbA_1c.
Conclusions Our results do not clearly support a causal link between exposure to cholinesterase-inhibiting insecticides and elevated blood glucose levels (expressed as HbA_1c and FPG), but results should be interpreted with caution due to the risk of reverse causality.

Chlorothalonil, a fungicide applied for decades worldwide, has recently been banned in the European Union (EU) and Switzerland due to its carcinogenicity and the presence of potentially toxic transformation products (TPs) in groundwater. The spread and concentration range of chlorothalonil TPs in different drinking water resources was examined (73 groundwater and four surface water samples mainly from Switzerland). The chlorothalonil sulfonic acid TPs (R471811, R419492, R417888) occurred more frequently and at higher concentrations (detected in 65-100% of the samples, ≤2200 ngL^-1) than the phenolic TPs (SYN507900, SYN548580, R611968; detected in 10-30% of the samples, ≤130 ngL^-1). The TP R471811 was found in all samples and even in 52% of the samples above 100 ngL^-1, the drinking water standard in Switzerland and other European countries. Therefore, the abatement of chlorothalonil TPs was investigated in laboratory and pilot-scale experiments and along the treatment train of various water works, comprising aquifer recharge, UV disinfection, ozonation, advanced oxidation processes (AOPs), activated carbon treatment, and reverse osmosis. The phenolic TPs can be abated during ozonation (second order rate constant k_O3 ∼10⁴ M^-1s^-1) and by reaction with hydroxyl radicals (OH) in AOPs (k_OH ∼10⁹ M^-1s^-1). In contrast, the sulfonic acid TPs, which occurred in higher concentrations in drinking water resources, react only very slowly with ozone (k_O3 <0.04 M^-1s^-1) and OH (k_OH <5.0 × 10⁷ M^-1s^-1) and therefore persist in ozonation and OH-based AOPs. Activated carbon retained the very polar TP R471811 only up to a specific throughput of 25 m³kg^-1 (20% breakthrough), similarly to the X-ray contrast agent diatrizoic acid. Reverse osmosis was capable of removing all chlorothalonil TPs by ≥98%.

Drains can be installed to reduce saturated soil conditions and improve crop growth. This article evaluates the extent to which these systems may cause losses of plant protection products (PPP). To that end, we estimated the drained fraction of Swiss arable land using both existing drainage maps and machine learning. Our drainage map suggests that 25% of Swiss agricultural land has a moderate to high potential of being drained. We further evaluated the risk for PPP losses via drainage and the potential of selected mitigation measures based on a conceptual model summarising the relevant scientific literature. Although drainage losses are highly variable, they are strongly influenced by the extent of preferential flow (PF) in soils and are an important transport pathway to surface waters. Most agricultural soils in Switzerland are prone to PF, given their loamy texture, suggesting that PPP losses via tile drains are an important phenomenon in Switzerland's drained arable fields. The most common mitigation measures for drainage are those recommended for runoff, erosion and leaching. However, given the uncertainty of the local PPP losses and driving factors, no site- or PPP-specific measures can currently be proposed to reduce PPP drainage losses. This study also examines the effectiveness of the regulatory model EXPOSIT in predicting total PPP losses and peak concentrations in watercourses. When compared with experimental data from Agroscope at Zurich-Affoltern, our findings suggest that EXPOSIT does not yield the worst-case estimates for Swiss conditions.

Bioaccumulation assessment predominantly relies on the bioconcentration factor (BCF) as sole decisive metric. The OECD Test Guideline number 305 (TG 305) provides the standard procedure for deriving this in vivo fish BCF, which requires a lot of animals, is expensive, and labor intensive. Accordingly, there is a lot of need and interest for alternative methods that can help to reduce, replace, and refine vertebrate tests as described in the 3R principles. Two alternative approaches have been developed in the past years: The bioconcentration test with the freshwater amphipod Hyalella azteca (HYBIT), and the OECD TG 319, provides a method to determine experimentally derived in vitro metabolism rates that can then be incorporated into in silico prediction models for rainbow trout BCF calculation. In the present study both alternative methods were applied to five substances of different physicochemical characteristics. The obtained results were compared with literature values of fish in vivo BCFs and additional BCFs obtained with the alternative methods, if available. Potential differences between the results of the test methods are discussed utilizing information such as in vivo metabolism rates. The currently available dataset suggests that both alternative methods pose promising alternatives to predict bioaccumulation in fish, although, defined applicability domains have yet to be determined.

Nitrification and distillation of urine allow for the recovery of all nutrients in a highly concentrated fertilizer solution. However, pharmaceuticals excreted with urine are only partially removed during these two process steps. For a sustainable and safe application, more extensive removal of pharmaceuticals is necessary. To enhance the pharmaceutical removal, which is already occurring during urine storage, nitrification and distillation, an adsorption column with granular activated carbon (GAC) can be included in the treatment train. We executed a pilot-scale study to investigate the adsorption of eleven indicator pharmaceuticals on GAC. During 74 days, we treated roughly 1000 L of pre-filtered and nitrified urine spiked with pharmaceuticals in two flow-through GAC columns filled with different grain sizes. We compared the performance of these columns by calculating the number of treated bed volumes until breakthrough and carbon usage rates. The eleven spiked pharmaceuticals were candesartan, carbamazepine, clarithromycin, diclofenac, emtricitabine, hydrochlorothiazide, irbesartan, metoprolol, N₄-acetylsulfamethoxazole, sulfamethoxazole and trimethoprim. At the shortest empty bed contact time (EBCT) of 25 min, immediate breakthrough was observed in both columns shortly after the start of the experiments. Strong competition by natural organic material (NOM) could have caused the low pharmaceutical removal at the EBCT of 25 min. At EBCTs of 70, 92 and 115 min, more than 660 bed volumes could be treated until breakthrough in the column with fine GAC. The earliest breakthrough was observed for candesartan and clarithromycin. On coarse GAC, only half the number of bed volumes could be treated until breakthrough compared to fine GAC. The probable reason for the later breakthrough with fine GAC is the smaller intraparticle diffusive path length. DOC and UV absorbance measurements at 265 nm indicated that both parameters can be used as indicators for the breakthrough of pharmaceuticals. In contrast to pharmaceuticals and DOC, the nutrient compounds ammonium, nitrate, phosphate, potassium and sulfate were not removed significantly. A comparison with literature values suggests that the amount of GAC needed to remove pharmaceuticals from human excreta could be reduced by nearly two orders of magnitude, if urine were treated on site instead of being discharged and treated in a centralized wastewater treatment plant.

The occurrence of legacy and novel per- and polyfluoroalkyl substances (PFASs) in multiple matrices from a farmland environment was investigated in the Beijing-Tianjin-Hebei core area of northern China. PFASs were ubiquitously detected in farmland soils, and the detection frequency of 6:2 chlorinated polyfluoroalkyl ether sulfonic acid (6:2 Cl-PFESA) was higher than that of perfluorooctane sulfonic acid (98% vs. 83%). Long-chain PFASs, including 6:2 Cl-PFESA, showed a centered distribution pattern around the metropolis of Tianjin, probably due to the local intensive industrial activity, while trifluoroacetic acid (TFA) showed a decreasing trend from the coast to the inland area. Other than soil, TFA was also found at higher levels than other longer-chain PFASs in dust, maize (Zea mays), poplar (Populus alba) leaf and locust (Locusta migratoria manilens) samples. Both poplar leaves and locusts can be used as promising biomonitoring targets for PFASs in farmland environments, and their accumulation potential corresponds with protein and lipid contents. Apart from being exposed to PFASs via food intake, locusts were likely exposed via uptake from soil and precipitated dust in farmland environments. The biomonitoring of locusts may be more relevant to insectivores, which is important to conducting a comprehensive ecological risk assessment of farmland environments.

Oceans are the ultimate sink for many of the over 100 million man-made substances. Until now, monitoring was limited to a reduced number of targeted persistent organic pollutants, reaching open waters mainly via atmospheric deposition. However, the composition and fate of the thousands of pollutants reaching the marine environment though wastewater discharges from coastal sources remain largely unexplored. By combining a newly developed nontarget screening (NTS) workflow and high-resolution mass spectrometry (HRMS), we have identified over 500 sewage-derived contaminants occurring in the ocean. Samples from the NE Atlantic contained this anthropogenic imprint at distances over 50 km from the coastline and >500 m depth, beyond the continental margin. The range of identified compounds spans from pharmaceuticals and personal care products to food additives and industrial chemicals, including several that have never been reported in the environment, as they escaped conventional targeted analytical methods. Predicting the effects of the continuous input of this chemical "cocktail" on marine ecosystems is a formidable challenge, since 40% of the detected compounds lack information regarding their use and ecotoxicity.

Hypochlorous acid (HOCl) is typically assumed to be the primary reactive species in free available chlorine (FAC) solutions. Lately, it has been shown that less abundant chlorine species such as chlorine monoxide (Cl₂O) and chlorine (Cl₂) can also influence the kinetics of the abatement of certain organic compounds during chlorination. In this study, the chlorination as well as bromination kinetics and mechanisms of 12 olefins (including 3 aliphatic and 9 aromatic olefins) with different structures were explored. HOCl shows a low reactivity towards the selected olefins with species-specific second-order rate constants <1.0 M⁻¹s⁻¹, about 4-6 orders of magnitude lower than those of Cl₂O and Cl₂. HOCl is the dominant chlorine species during chlorination of olefins under typical drinking water conditions, while Cl₂O and Cl₂ are likely to play important roles at high FAC concentration near circum-neutral pH (for Cl₂O) or at high Cl⁻ concentration under acidic conditions (for Cl₂). Bromination of the 12 olefins suggests that HOBr and Br₂O are the major reactive species at pH 7.5 with species-specific second-order rate constants of Br₂O nearly 3-4 orders of magnitude higher than of HOBr (ranging from <0.01 to >10³ M⁻¹s⁻¹). The reactivities of chlorine and bromine species towards olefins follow the order of HOCl < HOBr < Br₂O < Cl₂O ≈ Cl₂. Generally, electron-donating groups (e.g., CH₂OH- and CH₃-) enhances the reactivities of olefins towards chlorine and bromine species by a factor of 3-10², while electron-withdrawing groups (e.g., Cl-, Br-, NO₂-, COOH-, CHO-, -COOR, and CN-) reduce the reactivities by a factor of 3-10⁴. A reasonable linear free energy relationship (LFER) between the species-specific second-order rate constants of Br₂O or Cl₂O reactions with aromatic olefins and their Hammett σ⁺ was established with a more negative ρ value for Br₂O than for Cl₂O, indicating that Br₂O is more sensitive to substitution effects. Chlorinated products including HOCl-adducts and decarboxylated Cl-adduct were identified during chlorination of cinnamic acid by high-performance liquid chromatography/high resolution mass spectrometry (HPLC/HRMS).

The accurate and precise analysis of isotopologue and tandem mass isotopologue ratios in heavy stable isotope labeling experiments is a critical part of assessing absolute intracellular metabolic fluxes. Resulting from feeding the organism of interest with a specifically isotope-labeled substrate, the principal characteristics of these labeling experiments are the metabolites’ non-naturally distributed isotopologue patterns. For the purpose of inferring metabolic rates by maximizing the fit between a priori simulated and experimentally obtained labeling patterns, ¹³C is the preferred stable isotope of use.
The analysis of the obtained labeling patterns can be approached by different mass spectrometric approaches. Gas chromatography (GC) features broad metabolite coverage and excellent separation efficiency of biologically relevant isomers. These advantages compensate for laborious derivatization steps and the resulting need for interference correction for natural abundant isotopes.
Here, we describe a workflow based on GC-high resolution mass spectrometry with chemical ionization for the analysis of carbon-isotopologue distributions and some positional labeling information of primary metabolites. To study the associated measurement uncertainty of the resulting ¹³C labeling patterns, guidance to uncertainty estimation according to the EURACHEM guidelines with Monte-Carlo simulation is provided.

Wastewater treatment plant (WWTP) effluents release not only chemical constituents in watersheds, but also contain microorganisms. Thus, an understanding of what microorganisms are released and how they change microbial communities within natural streams is needed. To characterize the community shifts in streams receiving WWTP effluent, we sampled water-column microorganisms from upstream, downstream, and the effluent of WWTPs located on 23 headwater streams in which no other effluent was released upstream. We characterized the bacterial community by sequencing the V3-V4 region of the 16S rRNA gene. We hypothesized that the downstream community profile would be a hydraulic mixture between the two sources (i.e., upstream and effluent). In ordination analyses, the downstream bacterial community profile was a mixture between the upstream and effluent. For 14 of the sites, the main contribution (>50%) to the downstream community originated from bacteria in the WWTP effluent and significant shifts in relative abundance of specific sequence variants were detected. These shifts in sequence variants may serve as general bioindicators of wastewater-effluent influenced streams, with a human-gut related Ruminococcus genus displaying the highest shift (30-fold higher abundances downstream). However, not all taxa composition changes were predicted based on hydraulic mixing alone. Specifically, the decrease of Cyanobacteria/Chloroplast reads was not adequately described by hydraulic mixing. The potential alteration of stream microbial communities via a high inflow of human-gut related bacteria and a decrease in autotrophic functional groups resulting from WWTP effluent creates the potential for general shifts in stream ecosystem function.

Auf der Kläranlage Glarnerland wurde im Rahmen des Forschungsprojektes «Pilotversuche zur erweiterten Abwasserbehandlung mit granulierter Aktivkohle (GAK) und kombiniert mit Teilozonung (O3/GAK)» die Machbarkeit und Effizienz der genannten Verfahrenstechniken zur Spurenstoffelimination untersucht. Im weiteren Verlauf der Untersuchungen wurde zudem auch die Option der Dosierung von Pulveraktivkohle (PAK) in die biologische Stufe, welche mit dem S::Select®-Verfahren betrieben wird, als Vorstufe zur GAK erprobt.

Ziel der Pilotierung war die Erarbeitung von Entscheidungsgrundlagen zur Auswahl eines optimalen Verfahrens oder einer Verfahrenskombination für die angestrebte Realisierung der Spurenstoffelimination beim Abwasserverband Glarnerland. Zudem sollten Langzeiterfahrungen zum Betrieb von GAK-Filtrationen bezüglich Effizienz, Betrieb und vor allem Standzeiten (erreichte Bettvolumina) von verschiedenen GAK-Typen bei unterschiedlichen Kontaktzeiten bei einem Abwasser mit hohem gelöstem organischen Kohlenstoff (DOC) gesammelt werden.

Die Pilotanlage Vorozonung/GAK-Filtration ging Ende Januar 2017 auf der ARA Glarnerland in Betrieb und wurde im August 2019 ausser Betrieb gesetzt. Es wurden vier verschiedene GAK-Produkte untersucht (CSC «HC HK 1000», CarboTech «Pool W1-3», Jacobi «Aquasorb 5010», CabotNorit «GAC 1020 EN»). Parallel wurden alle GAK-Kolonnen (ausser Jacobi «Aquasorb 5010») während der gesamten Betriebsdauer mit einer Vorozonung bei einer spezifischen Ozondosis von rund 0.2 gO3/gDOC betrieben. [...]
 

Many organic contaminants entering the aquatic environment feature stereogenic structural elements that give rise to enantiomerism. While abiotic processes usually act identical on enantiomers, biotic processes, such as biodegradation often result in enantiomeric fractionation (EFr), i.e. the change of the relative abundance of enantiomers. Therefore, EFr offers the opportunity to differentiate biodegradation in complex environmental systems from abiotic processes. In this study, an achiral-chiral two-dimensional liquid chromatographic method for the enantioseparation of selected pharmaceuticals was developed. This method was then applied to determine the enantiomeric compositions of 8 chiral pharmaceuticals in 20 water-sediment test flumes and test EFr as an indicator of biodegradation. While all 8 substances were attenuated by at least 60%, 5 (atenolol, metoprolol, celiprolol, propranolol, flecainide) displayed EFr. No EFr was observed for citalopram, fluoxetine and venlafaxine despite almost complete attenuation (80 to 100%). Celiprolol, a barely studied β-blocker, revealed the most distinct EFr among all investigated substances, however, EFr varied considerably with biodiversity. Celiprolol-H2 was identified as a biological transformation product, possibly formed by reduction of the celiprolol keto group through a highly regio- and enantioselective carbonyl reductase. While celiprolol-H2 was observed across all flumes, as expected, its formation was faster in flumes with high bacterial diversity where also EFr was highest. Overall, EFr and transformation product formation together served as good indicators of biological processes; however, the strong dependence of EFr on biodiversity limits its usefulness in complex environmental systems.

Desphenylchloridazon (DPC), the main metabolite of the herbicide chloridazon (CLZ), is more water soluble and persistent than CLZ and frequently detected in water bodies. When assessing DPC transformation in the environment, results can be nonconclusive if based on concentration analysis alone because estimates may be confounded by simultaneous DPC formation from CLZ. This study investigated the fate of DPC by combining concentration-based methods with compound-specific C and N stable isotope analysis (CSIA). Additionally, DPC formation and transformation processes were experimentally deconvolved in a dedicated lysimeter study considering three scenarios. First, surface application of DPC enabled studying its degradation in the absence of CLZ. Here, CSIA provided evidence of two distinct DPC transformation processes: one shows significant changes only in ¹³C/¹²C, whereas the other involves changes in both ¹³C/¹²C and ¹⁵N/¹⁴N isotope ratios. Second, surface application of CLZ mimicked a realistic field scenario, showing that during DPC formation, ¹³C/¹²C ratios of DPC were depleted in ¹³C relative to CLZ, while ¹⁵N/¹⁴N ratios remained constant. Finally, CLZ depth injection simulated preferential flow and demonstrated the importance of the topsoil for retaining DPC. The combination of the lysimeter study with CSIA enabled insights into DPC transformation in the field that are superior to those of studies of concentration trends.

This study aimed to compare three approaches for predicting the service life of full-scale GAC adsorbers for the removal of micropollutants. The approaches included (i) rapid small-scale column tests (RSSCTs), (ii) two pilot-scale sampling approaches, and (iii) predictive correlations that consider micropollutant properties and background water matrix characteristics. The RSSCT could predict full-scale performance only if a micropollutant-specific fouling index was applied. At the pilot-scale, water samples were collected (1) over time at the top sampling point only (empty bed contact time (EBCT) of 1 minute) to minimize time to breakthrough (method 1) and (2) at different column depths at a single time point (method 2). Breakthrough curves obtained with method 2 more closely matched those obtained at the full-scale. In addition, method 2 is more convenient since it requires only one sampling campaign. Method 2 was used as a prognostic tool to predict breakthrough curves for micropollutants without full-scale data and a comparison with an existing prediction model gave satisfactory results for 6 out of 13 compounds.

Temperature is a key factor that influences chemical biotransformation potential and rates, on which exposure and fate models rely to predict the environmental (micro)pollutant fate. Arrhenius-based models are currently implemented in environmental exposure assessment to adapt biotransformation rates to actual temperatures, assuming validity in the 0-30 °C range. However, evidence on how temperature shifts affect the physicochemical and microbial features in biological systems is scarce, questioning the validity of the existing modeling approaches. In this work, laboratory-scale batch assays were designed to investigate how a mixed microbial community responds to short-term temperature shifts, and how this impacts its ability to biotransform a range of structurally diverse micropollutants. Our results revealed three distinct kinetic responses at temperatures above 20 °C, mostly deviating from the classic Arrhenius-type behavior. Micropollutants with similar temperature responses appeared to undergo mostly similar initial biotransformation reactions, with substitution-type reactions maintaining Arrhenius-type behavior up to higher temperatures than oxidation-type reactions. Above 20 °C, the microbial community also showed marked shifts in both composition and activity, which mostly correlated with the observed deviations from Arrhenius-type behavior, with compositional changes becoming a more relevant factor in biotransformations catalyzed by more specific enzymes (e.g., oxidation reactions). Our findings underline the need to re-examine and further develop current environmental fate models by integrating biological aspects, to improve accuracy in predicting the environmental fate of micropollutants.

Untreated sewer overflows can contaminate receiving waters with micropollutants. Although concentrations of discharged micropollutants can be ecotoxicologically relevant, only limited data is available to assess occurrence and spatial differences among sewer overflow catchments. Therefore, we present an innovative type of data obtained with passive samplers at 20 combined sewer overflow sites (2-7 events per site; 95 events in total). The data sheds light on concentration ranges for 13 representative polar organic micropollutants and shows that micropollutants in both municipal wastewater and stormwater can be relevant sources of contaminants. We identify indicator micropollutants for further studies: benzotriazole (80% interquantile of time-weighted average concentration: 250–4800 ng/L), carbamazepine (33-910 ng/L), diclofenac (78-1000 ng/L), carbendazim (21-900 ng/L), diazinon (2.1-53 ng/L), diuron (22-1100 ng/L), mecoprop (98–5300 ng/L), metolachlor (6-230 ng/L), and terbutryn (29-810 ng/L). These concentration estimates are assumed to be on the safe side for comparison with environmental quality standards (EQS). A majority of sewer overflow sites (13 of 20) show discharge concentrations above acute EQS for at least one micropollutant and thus would have to rely on dilution by receiving waters to not exceed any EQS. The intersite variability among sewer overflows exceed the within-site variability. Hence, future monitoring studies should cover more sewer overflow sites. No correlation could be found with event durations, specific storage volume or land use data, thus showing the complexity of micropollutant occurrence and indicating that other factors led to the observed high spatial variability. In conclusion, our results clearly show the potential relevance of micropollutants in sewer overflows and the need to assess site-specific measures.

Intensified cyanobacterial bloom events are of increasing global concern because of adverse effects associated with the release of bioactive compounds, including toxic cyanopeptides. Cyanobacteria can produce a variety of cyanopeptides, yet our knowledge about their abundance and co-production remains limited. We applied a suspect-screening approach, including 700 structurally known cyanopeptides, and identified 11 cyanopeptides in Microcystis aeruginosa and 17 in Dolichospermum flos-aquae. Total cyanopeptide concentrations ranged from high nmol to μmol g_dry^-1 with slightly higher cell quotas in the mid-exponential growth phase. Relative cyanopeptide profiles were unchanged throughout the growth cycle. We demonstrate that quantification based on microcystin-LR equivalents can introduce an error of up to 6-fold and recommend a class-equivalent approach instead. In M. aeruginosa, rarely studied cyclamides dominated (>80%) over cyanopeptolins and microcystins. While all nutrient reductions caused less growth, only lowering phosphorous and micronutrients reduced cyanopeptide production by M. aeruginosa. Similar trends were observed for D. flos-aquae and only lowering nitrogen decreased cyanopeptide production while the relative abundance of individual cyanopeptides remained stable. The synchronized production of other cyanopeptides along with microcystins emphasizes the need to make them available as reference standards to encourage more studies on their occurrence in blooms, persistence, and potential toxicity.

Hyporheic zones are the water-saturated flow-through subsurfaces of rivers which are characterized by the simultaneous occurrence of multiple physical, biological, and chemical processes. Two factors playing a role in the hyporheic attenuation of organic contaminants are sediment bedforms (a major driver of hyporheic exchange) and the composition of the sediment microbial community. How these factors act on the diverse range of organic contaminants encountered downstream from wastewater treatment plants is not well understood. To address this knowledge gap, we investigated dissipation half-lives (DT50s) of 31 substances (mainly pharmaceuticals) under different combinations of bacterial diversity and bedform-induced hyporheic flow using 20 recirculating flumes in a central composite face factorial design. By combining small-volume pore water sampling, targeted analysis, and suspect screening, along with quantitative real-time PCR and time-resolved amplicon Illumina MiSeq sequencing, we determined a comprehensive set of DT50s, associated bacterial communities, and microbial transformation products. The resulting DT50s of parent compounds ranged from 0.5 (fluoxetine) to 306 days (carbamazepine), with 20 substances responding significantly to bacterial diversity and four to both diversity and hyporheic flow. Bacterial taxa that were associated with biodegradation included Acidobacteria (groups 6, 17, and 22), Actinobacteria (Nocardioides and Illumatobacter), Bacteroidetes (Terrimonas and Flavobacterium) and diverse Proteobacteria (Pseudomonadaceae, Sphingomonadaceae, and Xanthomonadaceae). Notable were the formation of valsartan acid from irbesartan and valsartan, the persistence of N-desmethylvenlafaxine across all treatments, and the identification of biuret as a novel transformation product of metformin. Twelve additional target transformation products were identified, which were persistent in either pore or surface water of at least one treatment, indicating their environmental relevance.

Natural estrogens such as 17α-estradiol (E2α), 17β-estradiol (E2β), estrone (E1), and estriol (E3), released to surface waters from both urban and agricultural sources, are endocrine disrupting for fish. Here, we assess the prevalence of livestock farming derived natural estrogens in tributaries and ponds in the agriculturally dominated catchment of Lake Baldegg, Switzerland. Passive samplers were deployed in the main tributary and daily time-proportional water samples were collected in five tributaries for 30 days at the beginning of the vegetation period. Furthermore, we took grab samples of 12 ponds in the catchment. Aqueous samples were liquid–liquid extracted, derivatized, and analysed with LC-MS/MS and stream water samples additionally with ERα-CALUX, a bioassay for assessing total estrogenic activity. Natural estrogens were regularly detected, with mean concentrations ranging from below the limit of detection to 0.55 ng L⁻¹ for E2β and E1, respectively, and passive sampling and bioassay results largely confirmed these findings. Monte Carlo simulated mean natural estrogen concentrations underestimated measured ones by a factor of three to 11. An agricultural area's hydrological contribution and connectivity to surface waters seemed to be more important for the development of estrogen concentrations in streams than livestock densities in a catchment or the actual loads of slurry applied. Pond water occasionally contained natural estrogens in concentrations up to 8.6 ng L⁻¹ for E2α. The environmental quality standards of the European Union (0.4 ng L⁻¹ for E2β and 3.6 ng L⁻¹ for E1) were never exceeded for longer than a day in tributaries, but E1 reached critical concentrations for aquatic organisms in ponds.

Natural estrogens act as endocrine disruptors. However, the fate of livestock farming derived natural estrogens (17α-estradiol, 17β-estradiol, estrone, and estriol) in slurry is not well understood. In this study, we assessed the effects of on farm-storage on natural estrogen concentrations in slurry. Furthermore, we monitored pig and cattle slurry pits from major agricultural areas in Switzerland and determined natural estrogen concentrations therein. They were relatively stable over time, and mean concentrations ranged from 138 to 861 and 54 to 244 ng/L for cattle and pig slurries, respectively. 17α-Estradiol and estriol were the most prevalent estrogens in cattle and pig slurries, respectively. Based on livestock numbers, agricultural area, and estrogen concentrations in slurry, the estimated annual load of total natural estrogens applied on agricultural area amounted to 36 mg/ha. Our results indicate that slurry application is a relevant source of natural estrogens in the environment.

Conventional urban wastewater treatment plants (UWTPs) are poorly effective in the removal of most contaminants of emerging concern (CECs), including antibiotics, antibiotic resistant bacteria and antibiotic resistance genes (ARB&ARGs). These contaminants result in some concern for the environment and human health, in particular if UWTPs effluents are reused for crop irrigation. Recently, stakeholders' interest further increased in Europe, because the European Commission is currently developing a regulation on water reuse. Likely, conventional UWTPs will require additional advanced treatment steps to meet water quality limits yet to be officially established for wastewater reuse. Even though it seems that CECs will not be included in the proposed regulation, the aim of this paper is to provide a technical contribution to this discussion as well as to support stakeholders by recommending possible advanced treatment options, in particular with regard to the removal of CECs and ARB&ARGs. Taking into account the current knowledge and the precautionary principle, any new or revised water-related Directive should address such contaminants. Hence, this review paper gathers the efforts of a group of international experts, members of the NEREUS COST Action ES1403, who for three years have been constructively discussing the efficiency of the best available technologies (BATs) for urban wastewater treatment to abate CECs and ARB&ARGs. In particular, ozonation, activated carbon adsorption, chemical disinfectants, UV radiation, advanced oxidation processes (AOPs) and membrane filtration are discussed with regard to their capability to effectively remove CECs and ARB&ARGs, as well as their advantages and drawbacks. Moreover, a comparison among the above-mentioned processes is performed for CECs relevant for crop uptake. Finally, possible treatment trains including the above-discussed BATs are discussed, issuing end-use specific recommendations which will be useful to UWTPs managers to select the most suitable options to be implemented at their own facilities to successfully address wastewater reuse challenges.

Die schweizerische Agrarlandschaft weist ein dichtes Weg- und Strassennetz auf, dessen Entwässerung oft direkt in Oberflächengewässer führt. Es wirkt damit als hydraulischer Kurzschluss und als Transportpfad für Pflanzenschutzmittel (PSM). Felduntersuchungen und Modellrechnungen zeigen, dass gleich viel Ackerfläche über Kurzschlüsse mit einem Gewässer verbunden ist wie direkt angeschlossen ist. Zusätzlich wurden bedeutende PSM-Konzentrationen in Kurzschlüssen gemessen. Das weist darauf hin, dass Kurzschlüsse erheblich zur Belastung der Schweizer Gewässer mit PSM beitragen.

Chemicals' half-lives derived from biotransformation simulation studies are central metrics for persistence assessment in international regulatory frameworks. To determine the persistence of chemicals released to the aquatic environment, paradigm shifts in recent and ongoing revisions of chemical legislation assign increasing importance to OECD 309 simulation studies. OECD 309 studies were designed to target biotransformation in natural water (pelagic test) or in water amended with sediment (suspension test). Suspension tests bear several advantages over the pelagic test, most importantly, employing higher bacterial cell densities, which promote biotransformation of various chemicals at observable rates. However, experience with suspension tests is limited. In this study, we followed the fate of 43 pharmaceuticals, pesticides, and industrial chemicals in various suspension test setups and elucidated parameters influencing biotransformation kinetics and half-lives derived thereof. Besides striking intrastudy variability between replicates, we found that differences in sediment origin and bacterial cell density resulted in chemical half-lives that were different by up to 2 orders of magnitude, making persistence classification rather uncertain. However, data suggested that test systems employing bacterial cell densities close to the upper limit of what is commonly observed in natural surface waters (i.e., 107 cells mL-1) yielded increased and more uniform biotransformation of chemicals.

Wie synthetische Pestizide in die Gewässer gelangen und welche Auswirkungen das hat.

Pesticides are used globally in agriculture and pose a threat to the health of farmers, communities, and the environment. Smallholder farmers in low- and middle-income countries have generally a low socio-economic status and educational level. Consequently, they are particularly vulnerable to negative impacts of pesticides on their health, yields, or land. In a Knowledge, Attitude, and Practices study, we compared the pest management practices between a market-oriented farming system in Zarcero County, Costa Rica, and a subsistence-based farming system in Wakiso District, Uganda. We conducted a cross-sectional survey among smallholder farmers from Costa Rica (n = 300) in 2016 and from Uganda (n = 302) in 2017. We enrolled conventional and organic farmers, but also farmers with mixed practices and non-applicators of any pest management strategy. We found that the majority of pesticides used in both case studies are classified as highly hazardous by the World Health Organization. While more than 90% of smallholder farmers from both countries were aware of the negative health effects of pesticide exposure, <11% in Costa Rica and <2% in Uganda reported using personal protective equipment every time they handled or applied pesticides. Hygiene and other safe use practices were not adopted by all farmers (<61%), especially among farmers applying more hazardous pesticides. Conventional farmers from Costa Rica (14%) and Uganda (19%) reported disposing pesticide residuals into rivers. Using a logistic regression we found that organic farmers were more likely to having been trained on safe pesticide use practices. Using a robust regression, we observed that smallholder household income was primarily driven by education and not directly by the use of synthetic pesticides. Our results suggest that negative effects of pesticides can be managed over the whole life cycle, from purchase, via storage and application to residual and waste management by fostering professionalization of farmers. We advise future safe use and handling interventions to consider the pesticide use-related socioeconomic and demographic findings highlighted in this paper.

Five-membered nitrogen-containing heterocyclic compounds (azoles) belong to potential moieties in complex structures where transformations during ozonation can occur. This study focused on the azole-ozone chemistry of pyrrole, imidazole, and pyrazole as model compounds. Reaction kinetics and ozonation products were determined by kinetic and analytical methods including NMR, LC-HRMS/MS, HPLC-UV, and IC-MS. Analyses of reactive oxygen species (¹O₂, ^•OH, H₂O₂), quantum chemical computations (Gibbs energies), and kinetic simulations were used to further support the proposed reaction mechanisms. The species-specific second-order rate constants for the reactions of ozone with pyrrole and imidazole were (1.4 ± 1.1) × 10⁶ M^-1 s^-1 and (2.3 ± 0.1) × 10⁵ M^-1 s^-1, respectively. Pyrazole reacted more slowly with ozone at pH 7 (k_app = (5.6 ± 0.9) × 10¹ M^-1 s^-1). Maleimide was an identified product of pyrrole with a 34% yield. Together with other products, formate, formamide, and glyoxal, C and N mass balances of ~50% were achieved. Imidazole reacted with ozone to cyanate, formamide, and formate (~100% yields per transformed imidazole, respectively) with a closed mass balance. For pyrazole, a high ozone:pyrazole molar stoichiometry of 4.6 was found, suggesting the transformation products contributed to the over-stoichiometric consumption of ozone (e.g., hydroxypyrazoles). Glyoxal and formate were the only identified transformation products (C mass balance: 65%). Overall, the identified major products are suspected to hydrolyze and/or be biodegraded and thereby abated by a biological post-treatment typically following ozonation. However, as sub-structures of more complex compounds (e.g., micropollutants), they might be more persistent during the biological post-treatment.

Preventing and remedying fresh waters from chemical pollution is a fundamental societal and scientific challenge. With other nonchemical stressors potentially co-occurring, assessing the ecological consequences of reducing chemical loads in the environment is arduous. In this case study, we comparatively assessed the community structure, functions, and tolerance of stream biofilms to micropollutant mixtures extracted from deployed passive samplers at wastewater treatment plant effluents. These biofilms were growing up- and downstream of one upgraded and two nonupgraded wastewater treatment plants before being sampled for analyses. Our results showed a substantial decrease in micropollutant concentrations by 85%, as the result of upgrading the wastewater treatment plant at one of the sampling sites with activated carbon filtration. This decrease was positively correlated with a loss of community tolerance to micropollutants and the recovery of the community structure downstream of the effluent. On the other hand, downstream biofilms at the nonupgraded sites displayed higher tolerance to the extracts than the upstream biofilms. The observed higher tolerance was positively linked to micropollutant levels both in stream water and in biofilm samples, and to shifts in the community structure. Although more investigations of upgraded sites are needed, our findings point toward the suitability of using community tolerance for the retrospective assessment of the risks posed by micropollutants, to assess community recovery, and to relate effects to causes in complex environmental conditions.

The lack of fundamental insights on the fate of micropollutants during conventional activated sludge treatment in wastewater treatment plants (WWTPs) presents one of the biggest challenges in optimizing their removal. To address this challenge, we designed a study to identify the drivers of micropollutant removal in WWTPs. We calculated the removal efficiency of micropollutants across the activated sludge process of full-scale wastewater treatment plants from literature-reported data. Our final dataset consisted of 529 independent observations for 84 micropollutants along with a set of associated activated sludge process parameters. We used the Eawag pathway prediction system to predict initial biotransformations for each of the 84 micropollutants and hierarchical clustering to group chemicals based on similarities in their predicted initial sets of biotransformations. We then applied stability selection to generate well-performing models that can be interpreted to uncover the key factors contributing to the range of removal efficiencies calculated for each cluster of micropollutants. The key factors considered in stability selection included six physicochemical properties of the micropollutants and six activated sludge process parameters. The sludge-water partitioning coefficient, molecular weight, solids retention time, influent micropollutant concentration, and dissolved oxygen levels were consistently identified as key factors of micropollutant removal for clusters of micropollutants that undergo certain types of initial biotransformations. Our findings highlight the importance of considering initial biotransformations when evaluating micropollutant removal and identify important process parameters that determine the fate of micropollutants during activated sludge treatment.

Understanding the dynamics of groundwater resources subject to the pressures of human activity has become a central topic in 21st century hydrogeology. Groundwater availability is defined both in terms of quantity - how much water is stored an aquifer - and in terms of quality - how much groundwater of sufficient quality is available for consumption. Changes in land use have directly  impacted both of these facets. Due to its hidden nature, accurately determining the amount of available groundwater is notoriously difficult, and the added complexity of human stressors only augment these difficulties. In addition, the human landscape itself is undergoing rapid change, with a consistent increase in the human population, most of who now live in cities. This population increase has led to increasing land surface dedicated to anthropogenic use, and requires ever greater amounts of water resources for drinking water, agricultural applications, and industrial processes. Hence why a rising number of efforts have emerged in scientific literature on the topic of groundwater in the human environment. [...]

Das Verständnis der Dynamik von Grundwasserressourcen, die dem Druck menschlicher Aktivitäten ausgesetzt sind, ist zu einem zentralen Thema der Hydrogeologie des 21. Jahrhunderts geworden. Die Verfügbarkeit von Grundwasser wird sowohl quantitativ definiert - wie viel Wasser in einem Grundwasserleiter gespeichert ist - als auch qualitativ - wie viel Grundwasser in ausreichender Qualität für den Verbrauch zur Verfügung steht. Veränderungen in der Landnutzung haben sich direkt auf diese beiden Facetten ausgewirkt. Aufgrund seiner verborgenen Natur ist die genaue Bestimmung der Menge des verfügbaren Grundwassers bekanntermassen schwierig, und die zusätzliche Komplexität aus menschlichen Stressoren verstärkt diese Schwierigkeiten nur noch. Hinzu kommt, dass die menschliche Gesellschaft selbst einem raschen Wandel unterworfen ist, mit einem stetigen Anstieg der menschlichen Bevölkerung, von der grössere Teil heute in Städten lebt. Diese Bevölkerungszunahme hatte zu einer Vergrösserung der für die anthropogene Nutzung bestimmten Landfläche geführt und erfordert immer grössere Mengen an Wasserressourcen für Trinkwasser, landwirtschaftliche Anwendungen und industrielle Prozesse. Folglich sind in der wissenschaftlichen Literatur immer mehr Bemühungen zum Thema Grundwasser in der menschlichen Umwelt zu verzeichnen. [...]

Oxidative processes frequently contribute to organic pollutant degradation in natural and engineered systems, such as during the remediation of contaminated sites and in water treatment processes. Because a systematic characterization of abiotic reactions of organic pollutants with oxidants such as ozone or hydroxyl radicals by compound-specific stable isotope analysis (CSIA) is lacking, stable isotope-based approaches have rarely been applied for the elucidation of mechanisms of such transformations. Here, we investigated the carbon isotope fractionation associated with the oxidation of benzene and several methylated and methoxylated analogs, namely, toluene, three xylene isomers, mesitylene, and anisole, and determined their carbon isotope enrichments factors (ε_C) for reactions with ozone (ε_C = -3.6 to -4.6 ‰) and hydroxyl radicals (ε_C = 0.0 to -1.2‰). The differences in isotope fractionation can be used to elucidate the contribution of the reactions with ozone or hydroxyl radicals to overall transformation. Derivation of apparent kinetic isotope effects (AKIEs) for the reaction with ozone, however, was nontrivial due to challenges in assigning reactive positions in the probe compounds for the monodentate attack leading to an ozone adduct. We present several options for this step and compare the outcome to quantum chemical characterizations of ozone adducts. Our data show that a general assignment of reactive positions for reactions of ozone with aromatic carbons in ortho-, meta-, or para-positions is not feasible and that AKIEs of this reaction should be derived on a compound-by-compound basis.

New antimicrobial peptides are emerging as promising alternatives to conventional antibiotics because of their specificity for target pathogens and their potential to be rapidly hydrolyzed (i.e., inactivated) by extracellular peptidases during biological wastewater treatment, thereby limiting the emergence and propagation of antibiotic resistance in the environment. However, little is known about the specificity of extracellular peptidases derived from wastewater microbial communities, which is a major impediment for the design of sustainable peptide-based antibiotics that can be hydrolyzed by wastewater peptidases. We used a set of natural peptides to explore the specificity of dissolved extracellular wastewater peptidases. We found that enzyme-catalyzed hydrolysis occurred at specific sites and that a subset of the these hydrolyses were conserved across enzyme pools derived from three independent wastewater microbial communities. An analysis of the amino-acid residues flanking the hydrolyzed bonds revealed a set of residue motifs that were linked to enzyme-catalyzed hydrolysis and are therefore candidates for incorporation into new and sustainable peptide-based antibiotics.

Ferrous iron formed during microbial ferric iron reduction induces phase transformations of poorly crystalline into more crystalline and thermodynamically more stable iron (oxyhydr)oxides. Yet, characterizing the resulting decreases in the reactivity of the remaining oxide ferric iron toward reduction (i.e., its reducibility) has been challenging. Here, we used the reduction of six-line ferrihydrite by Shewanella oneidensis MR-1 as a model system to demonstrate that mediated electrochemical reduction (MER) allows directly following decreases in oxide ferric iron reducibility during the transformation of ferrihydrite into goethite and magnetite which we characterized by X-ray diffraction analysis and transmission electron microscopy imaging. Ferrihydrite was fully reducible in MER at both pH_MER of 5.0 and 7.5. Decreases in iron oxide reducibility associated with ferrihydrite transformation into magnetite were accessible at both pH_MER because the formed magnetite was not reducible under either of these conditions. Conversely, decreases in iron oxide reducibility associated with goethite formation were apparent only at the highest tested pH_MER of 7.5 and thus the thermodynamically least favorable conditions for iron oxide reductive dissolution. The unique capability to adjust the thermodynamic boundary conditions in MER to the specific reducibilities of individual iron (oxyhydr)oxides makes this electrochemical approach broadly applicable for studying changes in iron oxide reducibility in heterogeneous environmental samples such as soils and sediments.

Electron transfer to ferric iron in (oxyhydr-)oxides (hereafter iron oxides) is a critical step in many processes that are central to the biogeochemical cycling of elements and to pollutant dynamics. Understanding these processes requires analytical approaches that allow for characterizing the reactivity of iron oxides toward reduction under controlled thermodynamic boundary conditions. Here, we used mediated electrochemical reduction (MER) to follow changes in iron oxide reduction extents and rates during abiotic ferrous iron-induced transformation of six-line ferrihydrite. Transformation experiments (10 mM ferrihydrite-Fe^III) were conducted over a range of solution conditions (pH_trans = 6.50 to 7.50 at 5 mM Fe²⁺ and for pH_trans = 7.00 also at 1 mM Fe²⁺) that resulted in the transformation of ferrihydrite into thermodynamically more stable goethite or magnetite. The changes in iron oxide mineralogy during the transformations were quantified using X-ray diffraction analysis. MER measurements on iron oxide suspension aliquots collected during the transformations were performed over a range of pH_MER at constant applied reduction potential. The extents and rates of iron oxide reduction in MER decreased with decreasing reaction driving force resulting from both increasing pH_MER and increasing transformation of ferrihydrite into thermodynamically more stable iron oxides. We show that the decreases in iron oxide reduction extents and rates during ferrihydrite transformations can be linked to the concurrent changes in iron oxide mineralogy.

The historic emissions of polar micropollutants in a natural drinking water source were investigated by non-target screening with high-resolution mass spectrometry and open cheminformatics tools. The study area consisted of a riverbank filtration transect fed by the river Lek, a branch of the lower Rhine, and exhibiting up to 60-year travel time. More than 18,000 profiles were detected. Hierarchical clustering revealed that 43% of the 15 most populated clusters were characterised by intensity trends with maxima in the 1990s, reflecting intensified human activities, wastewater treatment plant upgrades and regulation in the Rhine riparian countries. Tentative structure annotation was performed using automated in silico fragmentation. Candidate structures retrieved from ChemSpider were scored based on the fit of the in silico fragments to the experimental tandem mass spectra, similarity to openly accessible accurate mass spectra, associated metadata and presence in a suspect list. 67 unique structures (72 over both ionisation modes) were tentatively identified, 25 of which were confirmed and included contaminants so far unknown to occur in bank filtrate or in natural waters at all, such as tetramethylsulfamide. This study demonstrates that many classes of hydrophilic organics enter riverbank filtration systems, persisting and migrating for decades if biogeochemical conditions are stable.

Environmental water quality monitoring aims to provide the data required for safeguarding the environment against adverse biological effects from multiple chemical contamination arising from anthropogenic diffuse emissions and point sources. Here, we integrate the experience of the international EU-funded project SOLUTIONS to shift the focus of water monitoring from a few legacy chemicals to complex chemical mixtures, and to identify relevant drivers of toxic effects. Monitoring serves a range of purposes, from control of chemical and ecological status compliance to safeguarding specific water uses, such as drinking water abstraction. Various water sampling techniques, chemical target, suspect and non-target analyses as well as an array of in vitro, in vivo and in situ bioanalytical methods were advanced to improve monitoring of water contamination. Major improvements for broader applicability include tailored sampling techniques, screening and identification techniques for a broader and more diverse set of chemicals, higher detection sensitivity, standardized protocols for chemical, toxicological, and ecological assessments combined with systematic evidence evaluation techniques. No single method or combination of methods is able to meet all divergent monitoring purposes. Current monitoring approaches tend to emphasize either targeted exposure or effect detection. Here, we argue that, irrespective of the specific purpose, assessment of monitoring results would benefit substantially from obtaining and linking information on the occurrence of both chemicals and potentially adverse biological effects. In this paper, we specify the information required to: (1) identify relevant contaminants, (2) assess the impact of contamination in aquatic ecosystems, or (3) quantify cause–effect relationships between contaminants and adverse effects. Specific strategies to link chemical and bioanalytical information are outlined for each of these distinct goals. These strategies have been developed and explored using case studies in the Danube and Rhine river basins as well as for rivers of the Iberian Peninsula. Current water quality assessment suffers from biases resulting from differences in approaches and associated uncertainty analyses. While exposure approaches tend to ignore data gaps (i.e., missing contaminants), effect-based approaches penalize data gaps with increased uncertainty factors. This integrated work suggests systematic ways to deal with mixture exposures and combined effects in a more balanced way, and thus provides guidance for future tailored environmental monitoring.

A platform for archiving liquid chromatography high-resolution mass spectrometry (LC-HRMS) data was developed for the retrospective suspect screening of thousands of environmental pollutants with the ambition of becoming a European and possibly global standard. It was termed Digital Sample Freezing Platform (DSFP) and incorporates all the recent developments in the HRMS screening methods within the NORMAN Network. In the workflow, raw mass spectral data are converted into mzML, then mass spectral and chromatographic information on thousands of peaks of each sample is extracted into Data Collection Templates. The 'digitally frozen' samples can be retrospectively screened for the presence of virtually any compound amenable to LC–MS using a combination of information on its (i) exact mass, (ii) predicted retention time window in the chromatogram, (iii) isotopic fit and (iv) qualifier fragment ions. Its potential was demonstrated on monitoring of 670 antibiotics and 777 REACH chemicals from the Joint Black Sea Surveys (JBSS).

Peak prioritization plays a key role in non-target analysis of complex samples in order to focus the elucidation efforts on potentially relevant substances. The present work shows the development of a computational workflow capable of detecting compounds that exhibit large variation in intensity over time. The developed approach is based on three open-source R packages (xcms, CAMERA and TIMECOURSE) and includes the use of the statistical test Multivariate Empirical Bayes Approach to rank the compounds based on the Hotelling T2 coefficient, which is an indicator of large concentration variations of unknown components. The approach was applied to replicate series of 24 h composite flow-proportional influent wastewater samples collected during 8 consecutive days. 60 events involving unknown substances with high fluctuation over time were successfully prioritized. 14 of those compounds were tentatively identified using HRMS/MS libraries, chemical databases, in-silico fragmentation tools, and retention time prediction models. Four compounds were confirmed with standards from which two never reported before in wastewater.

Diatoms are among the most productive and ecologically important groups of microalgae in contemporary oceans. Due to their distinctive metabolic and physiological features, they offer exciting opportunities for a broad range of commercial and industrial applications. One such feature is their ability to synthesize a wide diversity of isoprenoid compounds. However, limited understanding of how these molecules are synthesized have until recently hindered their exploitation. Following comprehensive genomic and transcriptomic analysis of various diatom species, the biosynthetic mechanisms and regulation of the different branches of the pathway are now beginning to be elucidated. In this review, we provide a summary of the recent advances in understanding diatom isoprenoid synthesis and discuss the exploitation potential of diatoms as chassis for high-value isoprenoid synthesis.

Climate change is one of the largest and possibly most impactful ongoing but also future environmental drivers. Understanding effects of climate change on aquatic ecosystems and their function is thus of high importance. Within the framework of the Hydro-CH2018 project, the Swiss Federal Office for the Environment (FOEN) initiated a synthesis of climate change effects on water quality and the ecological status of freshwater ecosystems. The present report is the result of these synthesis efforts.

Our goal was to summarize available knowledge on how climatic change affects freshwater ecosystems in Switzerland. The work was performed between fall 2017 and fall 2018. We aimed to provide a synthesis of existing knowledge that is as consistent and complete as possible, but also to identify possible knowledge gaps. The work is based on extensive literature surveys and expert interviews. Whenever possible, we included quantitative conclusions. At the same time, we also wanted to be specific and relevant for natural Swiss water systems, so we also included qualitative effects and case studies. While many of the major ongoing and expected effects of climate change on aquatic systems are already well known, we also faced several challenges. For example: (i) some effects of climate change are still only partly understood, (ii) many of the focal endpoints, both with respect to water quality and ecological status, are not only affected by climate change, but also by other anthropogenic drivers, and disentangling these drivers is challenging if not infeasible, (iii) specific effects can be dependent on specific local factors, and (iv) the research field is so vast that the available resources for this one-year synthesis did not allow us to go into as much detail and data analysis as the topic would deserve. As a result, our report is a snapshot of scientific knowledge, at the period of realization, of the effects of climate change on aquatic ecosystems in Switzerland, set into the context of other environmental drivers.

While we see that climate change will have major effects on Swiss water systems, we also identify possible ways of mitigating these effects. We thereby hope that our synthesis is not only an overview of current and future changes, but will also help in policy- and decisionmaking about how to best deal with these changes and challenges.

The recent development of insensitive munitions, such as 2,4-dinitroanisole (DNAN), as components of military explosives has generated concern for potential subsurface contamination and created a need to fully characterize their transformation processes. Compound specific isotope analysis (CSIA) has proven to be a useful means of assessing transformation pathways according to characteristic stable isotope fractionation patterns. The C and N isotope fractionation of DNAN associated with abiotic and enzymatic hydrolysis was recently assessed. The extent to which DNAN isotope fractionation will be affected by other potentially competing transformation pathways known for nitroaromatic compounds (e.g., reduction) and if previous knowledge can be extrapolated to other environmental matrices remains to be understood. Here, we investigated the C and N isotope fractionation and reaction rate constants of DNAN during abiotic reduction mediated by mineral-associated Fe(II) species as a function of mineral type, natural organic matter presence, and repeated exposures to DNAN. Though rate constants varied, N and C apparent kinetic isotope effects (AKIEs) remained consistent across all experiments (averaged values of ¹⁵N-AKIE = 1.0317 ± 0.0064 and ¹³C-AKIE = 1.0008 ± 0.0005) and revealed significant ¹⁵N- and minimal ¹³C-enrichment in agreement with previous work on nitroaromatic compounds. Moreover, the observed fractionation was clearly distinct from trends for abiotic and enzymatic hydrolysis. This study provides a strengthened basis for the use of CSIA as a robust tool for monitoring DNAN degradation in complex environmental matrices as a component of future remediation efforts.

The present monitoring and assessment of the chemical status of water bodies fail to characterize the likelihood that complex mixtures of chemicals affect water quality. The European Collaborative Project SOLUTIONS suggests that this likelihood can be estimated with effect-based methods (EBMs) complemented by chemical screening and/or impact modeling. These methods should be used to identify the causes of impacted water quality and to develop programs of measures to improve water quality. Along this line of reasoning, effect-based methods are recommended for Water Framework Directive (WFD) monitoring to cover the major modes of action in the universe of environmentally relevant chemicals so as to evaluate improvements of water quality upon implementing the measures. To this end, a minimum battery of bioassays has been recommended including short-term toxicity to algae, Daphnia and fish embryos complemented with in vitro and short-term in vivo tests on mode-of-action specific effects as proxies for long-term toxicity. The likelihood of adverse impacts can be established with effect-based trigger values, which differentiate good from poor water quality in close alignment with Environmental Quality Standards for individual chemicals, while taking into account mixture toxicity. The use of EBMs is suggested in the WFD as one avenue to establish the likelihood of adverse effects due to chemical pollution in European water systems. The present paper has been written as one component of a series of policy briefs to support decisions on water quality monitoring and management under the WFD.

Currently, chemical monitoring based on priority substances fails to consider the majority of known environmental micropollutants not to mention the unexpected and unknown chemicals that may contribute to the toxic risk of complex mixtures present in the environment. Complementing component- and effect-based monitoring with wide-scope target, suspect, and non-target screening (NTS) based on high-resolution mass spectrometry (HRMS) data is recommended to support environmental impact and risk assessment. This will allow for detection of newly emerging compounds and transformation products, retrospective monitoring efforts, and the identification of possible drivers of toxicity by correlation with effects or modelling of expected effects for future and abatement scenarios. HRMS is becoming increasingly available in many laboratories. Thus, the time is right to establish and harmonize screening methods, train staff, and record HRMS data for samples from regular monitoring events and surveys. This will strongly enhance the value of chemical monitoring data for evaluating complex chemical pollution problems, at limited additional costs. Collaboration and data exchange on a European-to-global scale is essential to maximize the benefit of chemical screening. Freely accessible data platforms, inter-laboratory trials, and the involvement of international partners and networks are recommended.

To meet the United Nations (UN) sustainable development goals and the European Union (EU) strategy for a non-toxic environment, water resources and ecosystems management require cost-efficient solutions for prevailing complex contamination and multiple stressor exposures. For the protection of water resources under global change conditions, specific research needs for prediction, monitoring, assessment and abatement of multiple stressors emerge with respect to maintaining human needs, biodiversity, and ecosystem services. Collaborative European research seems an ideal instrument to mobilize the required transdisciplinary scientific support and tackle the large-scale dimension and develop options required for implementation of European policies. Calls for research on minimizing society’s chemical footprints in the water–food–energy–security nexus are required. European research should be complemented with targeted national scientific funding to address specific transformation pathways and support the evaluation, demonstration and implementation of novel approaches on regional scales. The foreseeable pressure developments due to demographic, economic and climate changes require solution-oriented thinking, focusing on the assessment of sustainable abatement options and transformation pathways rather than on status evaluation. Stakeholder involvement is a key success factor in collaborative projects as it allows capturing added value, to address other levels of complexity, and find smarter solutions by synthesizing scientific evidence, integrating governance issues, and addressing transition pathways. This increases the chances of closing the value chain by implementing novel solutions. For the water quality topic, the interacting European collaborative projects SOLUTIONS, MARS and GLOBAQUA and the NORMAN network provide best practice examples for successful applied collaborative research including multi-stakeholder involvement. They provided innovative conceptual, modelling and instrumental options for future monitoring and management of chemical mixtures and multiple stressors in European water resources. Advancement of EU water framework directive-related policies has therefore become an option.

Aim: The complications that arise when performing meta-analysis of datasets from multiple metabolomics studies are addressed with computational methods that ensure data quality, completeness of metadata and accurate interpretation across studies. Results & methodology: This paper presents an integrated system of quality control (QC) methods to assess metabolomics results by evaluating the data acquisition strategies and metabolite identification process when integrating datasets for meta-analysis. An ontology knowledge base and a rule-based system representing the experiment and chemical background information direct the processes involved in data integration and QC verification. A diabetes meta-analysis study using these QC methods finds putative biomarkers that differ between cohorts. Conclusion: The methods presented here ensure the validity of meta-analysis when integrating data from different metabolic profiling studies.

Water pollution is ubiquitous globally, yet how the effects of pollutants propagate through natural ecosystems remains poorly understood. This is because the interactive effects of multiple stressors are generally hard to predict. Agriculture and municipal wastewater treatment plants (WWTPs) are often major sources of contaminants for streams, but their relative importance and the role of different pollutants (e.g. nutrients or pesticides) are largely unknown. Using a 'real world experiment' with sampling locations up- and downstream of WWTPs, we studied how effluent discharges affected water quality and macroinvertebrate communities in 23 Swiss streams across a broad land-use gradient.
Variation partitioning of community composition revealed that overall water quality explained approximately 30% of community variability, whereby nutrients and pesticides each independently explained 10% and 2%, respectively. Excluding oligochaetes (which were highly abundant downstream of the WWTPs) from the analyses, resulted in a relatively stronger influence (3%) of pesticides on the macroinvertebrate community composition, whereas nutrients had no influence. Generally, the macroinvertebrate community composition downstream of the WWTPs strongly reflected the upstream conditions, likely due to a combination of efficient treatment processes, environmental filtering and organismal dispersal. Wastewater impacts were most prominently by the Saprobic index, whereas the SPEAR index (a trait-based macroinvertebrate metrics reflecting sensitivity to pesticides) revealed a strong impact of arable cropping but only a weak impact of wastewater.
Overall, our results indicate that agriculture can have a stronger impact on headwater stream macroinvertebrate communities than discharges from WWTP. Yet, effects of wastewater-born micropollutants were clearly quantifiable among all other influence factors. Improving our ability to further quantify the impacts of micropollutants requires highly-resolved water quality and taxonomic data with adequate spatial and temporal sampling. These improvements would help to better account for the underlying causal pathways that drive observed biological responses, such as episodic contaminant peaks and dispersal-related processes.

Polybrominated diphenyl ethers (PBDEs) are ubiquitous and toxic contaminants found in high concentrations in watercourses, and are not well removed by conventional wastewater treatment facilities. This study aimed to evaluate the removal and transformation of BDE-47, one of the environmentally predominant PBDE congener, by a green alga (Chlorella vulgaris) and a cyanobacterium (Microcystis flos-aquae) under different light conditions. Living and autoclaved cultures were exposed to BDE-47 at a concentration of 10 μg L⁻¹ for 7 days. Both species removed >90% of BDE-47 very shortly after spiking. Light intensity affected the transformation of BDE-47 in living cultures of both species, since 5 to 11 times more debromination products were measured at a light intensity of 100 μmol photons m⁻² s⁻¹ than at 20 μmol photons m⁻² s⁻¹. Living cultures of M. flos-aquae transformed BDE-47 at a rate of 0.22 day⁻¹ while no transformation was observed in the respective autoclaved cultures. On the contrary, both living and autoclaved cultures of C. vulgaris had similar BDE-47 transformation rates of 0.05–0.06 day⁻¹. Debromination of BDE-47 was a predominant transformation pathway in cultures of C. vulgaris, with two times higher BDE-28 concentrations measured than in M. flos-aquae, while hydroxylation was more dominant with the cyanobacterium. Most BDE-47 and its debromination product BDE-28 were found on the cell surface of both species. These results reveal that different transformation mechanisms were involved in C. vulgaris and M. flos-aquae cultures and confirm the importance of species selection for the removal of PBDEs from contaminated environments.

Cette synthèse de deux projets 62a illustre à l’aide d’exemples les effets des mesures de réduction des risques liés aux produits phytosanitaires (PPh) sur les données du monitoring. Au vu des multiples facteurs d’influence externes aux projets, ce lien entre les mesures prises par les agriculteurs et les données chimiques, biologiques et écotoxicologiques est parfois difficile à caractériser. Néanmoins, l’analyse des données de ces deux projets pilotes en Suisse permet de tirer de nombreuses leçons pour les projets en cours ou futurs, ainsi que pour le plan d’action PPh.

The clock is ticking and just a little more than 10 years remain to meet the 17 Sustainable Development Goals (SDGs). Enhanced political and public awareness, inter- and transdisciplinary engagement, new partnerships, and multisectoral collaborations are required to foster knowledge-and-action societies in order to tackle the complex issues that are inherent to sustainable development. A one-day symposium held in Basel, Switzerland, in November 2018 offered a venue for open exchange on how to stimulate dialogue for co-creating innovative ideas and scalable action to address some of the most pressing challenges of the 2030 Agenda for Sustainable Development.

Poor efficiency of TiO₂ by visible light and the fast recombination rate of electron/hole pairs remain significant challenges in photocatalytic applications of TiO₂ in water treatment. In this study, graphene was used to enhance TiO₂ photocatalytic activity by reducing electron-hole pair recombination. Nanocomposites were formed by combining TiO₂ nanoparticles (P25) with graphene oxide (GO) though simultaneous hydrothermal synthesis and GO reduction. Nanocomposite characterization confirmed that the GO was successfully reduced and P25 nanoparticles evenly dispersed in the graphene surface. The band gap of the nanocomposite was determined to be 2.74 eV, which is a promising shift to the visible spectrum in graphene-TiO₂ photocatalysts. The photocatalytic performance of the TiO₂/G nanocomposites was then evaluated by quantifying the formation of 2-hydroxyterephthalic acid (HTPA) (probe molecule) under UV-LED illumination. To further optimize the photocatalytic efficiency of the TiO₂/G nanocomposites, the response surface methodology (RSM) with central composite design (CCD) was used. Out of the 6 variables including stirring time, stirring speed, the amount of TiO₂, the amount of GO, hydrothermal reaction time, and ethanol/water ratio, it was determined that the last three are substantially affected the HTPA formation rate. The optimum conditions were found to be GO 0.48 wt%, ethanol/water 51.49 v/V%, and a reaction time 19 h. Predicted values for HTPA formation were found to be in good agreement with experimental values (R² = 0.93 and adj-R² = 0.87). The optimized nanocomposite showed 125% enhancement in photocatalytic efficiency over pure P25.

With the growth in production and use of chemicals and the fact that many end up in the aquatic environment, there is an increasing need for advanced water treatment technologies that can remove chemicals of emerging concern (CECs) from water. The current lack of a homogenous approach for testing advanced water treatment technologies hampers the interpretation and evaluation of CEC removal efficiency data, and hinders informed decision making by stakeholders with regard to which treatment technology could satisfy their specific needs.
Here a data evaluation framework is proposed to improve the use of current knowledge in the field of advanced water treatment technologies for drinking water and wastewater, consisting of a set of 9 relevance criteria and 51 reliability criteria. The two criteria sets underpin a thorough, unbiased and standardised method to select studies to evaluate and compare CEC removal efficiency of advanced water treatment technologies in a scientifically sound way.
The relevance criteria set was applied to 244 papers on removal efficiency, of which only 20% fulfilled the criteria. The reliability criteria were applied to the remaining papers. In general these criteria were fulfilled with regards to information on the target compound, the water matrix and the treatment process conditions. However, there was a lack of information on data interpretation and statistics.
In conclusion, a minority of the evaluated papers are suited for comparison across techniques, compounds and water matrixes. There is a clear need for more uniform reporting of water treatment studies for CEC removal. In the future this will benefit the selection of appropriate technologies.

Irrigation with treate wastewater (TWW) an application of biosolis introuce numerous pharmaceutical an personal care proucts (PPCPs) into agro-foo systems. While the use of TWW an biosolis has many societal benefits, introuction of PPCPs in prouction agriculture poses potential foo safety an human health risks. A comprehensive risk assessment an management scheme of PPCPs in agro-foo systems is limite by multiple factors, not least the sheer number of investigate compouns an their iverse structures. Here we follow the fate of PPCPs in the water-soil-prouce continuum by consiering processes an variables that influence PPCP transfer an accumulation. By analyzing the steps in the soil-plant-human iet nexus, we propose a tiere framework as a path forwar to prioritize PPCPs that coul have a high potential for plant accumulation an thus pose greatest risk. This article examines research progress to ate an current research challenges, highlighting the potential value of leveraging existing knowlege from ecaes of research on other chemicals such as pesticies. A process-riven scheme is outline to erive a short list that may be use to refocus our future research efforts on PPCPs an other analogous emerging contaminants in agro-foo systems.

Background: Pesticide use is increasing in low- and middle-income countries (LMICs) including Costa Rica. This increase poses health risks to farm owners, farm workers, and communities living near agricultural farms.

Objective: We aimed to examine the health effects associated with occupational pesticide exposure in farm owners and workers from conventional and organic smallholder farms in Costa Rica.

Methods: We conducted a cross-sectional study involving 300 owners and workers from organic and conventional horticultural smallholder farms in Zarcero County, Costa Rica. During the baseline study visit, we administered a structured, tablet-based questionnaire to collect data on sociodemographic characteristics, pesticide exposure, and health conditions (eg, respiratory and allergic outcomes and acute pesticide intoxication symptoms) and administered a neurobehavioral test battery (eg, Finger Tapping Test and Purdue Pegboard); we measured blood pressure, anthropometry (height, weight, and waist circumference), and erythrocytic acetylcholinesterase activity and also collected urine samples. In addition, a functional neuroimaging assessment using near-infrared spectroscopy was conducted with a subset of 50 study participants. During the follow-up study visit (~2-4 weeks after the baseline), we administered participants a short questionnaire on recent pesticide exposure and farming practices and collected hair, toenail, and urine samples. Urine samples will be analyzed for various pesticide metabolites, whereas toenails and hair will be analyzed for manganese (Mn), a biomarker of exposure to Mn-containing fungicides. Self-reported pesticide exposure data will be used to develop exposure intensity scores using an exposure algorithm. Furthermore, exposure-outcome associations will be examined using linear and logistic mixed-effects regression models.

Results: Fieldwork for our study was conducted between May 2016 and August 2016. In total, 113 farm owners and 187 workers from 9 organic and 83 conventional horticultural smallholder farms were enrolled. Data analyses are ongoing and expected to be published between 2019 and 2020.

Conclusions: This study is one of the first to examine differences in health effects due to pesticide exposure between farm owners and workers from organic and conventional smallholder farms in an LMIC. We expect that this study will provide critical data on farming practices, exposure pathways, and how occupational exposure to pesticides may affect farm owners and workers’ health. Finally, we hope that this study will allow us to identify strategies to reduce pesticide exposure in farm owners and workers and will potentially lay the groundwork for a future longitudinal study of health outcomes in farm owners and workers exposed to pesticides.

Wastewater treatment plant (WWTP) effluents release complex mixtures of organic and inorganic micropollutants, including endocrine disrupting compounds, into receiving water bodies. These substances may cause adverse effects in aquatic communities as well as in ecosystem functions they provide. The aim of this study was to determine the potential impact of secondary treated wastewater released into a small Swiss stream on leaf litter decomposition based on feeding rates of the amphipod shredder Gammarus fossarum measured in situ. Additionally, endocrine disrupting effects downstream of the WWTP were investigated by measuring vitellogenin (vg) induction in male gammarids exposed in situ, as well as estrogen receptor activation using the Yeast Estrogen Screen (YES) involving passive sampler and grab water sample extracts. Extracts were also analysed for 424 organic micropollutants and selected transformation products. Gammarid feeding rate was significantly reduced 100, 200 and 400 m downstream of the WWTP effluent relative to the upstream site. While YES results showed significantly elevated estrogenicity at downstream sites, vg production in male gammarids was not induced. A laboratory experiment, in which gammarids were exposed to WWTP effluent, supported this observation. These results, hence, suggest that treated wastewater released into aquatic ecosystems impairs the ecosystem function of leaf litter decomposition. Vg levels in male gammarids measured by UPLC-MS/MS did, however, not alter.

Aqueous film-forming foams (AFFFs) are proprietary mixtures containing hydrocarbon surfactants and per- and polyfluoroalkyl substances (PFASs) that are used to extinguish hydrocarbon-based fuel fires. There is limited information on hydrocarbon surfactants in AFFFs and AFFF-contaminated groundwater even though hydrocarbon surfactants are more abundant (5–10% w/w) than PFASs (0.9–1.5% w/w) in AFFFs. Eight commercial AFFFs manufactured between 1988 and 2012 and 10 AFFF-contaminated groundwaters collected from near source zones of fire-fighter training areas were analyzed for suspect hydrocarbon surfactants by liquid chromatography quadrupole time-of-flight mass spectrometry. A suspect list and a homologous series detection computational tool, enviMass, were combined to screen for hydrocarbon surfactants. Nine classes of hydrocarbon surfactants were detected in AFFFs including octylphenol polyethoxylates, linear alcohol ethoxylates, ethoxylated cocoamines, alkyl ether sulfates, alkyl amido dipropionates, linear alkyl benzenesulfonates, alkyl sulfates, and polyethylene glycols. Of those, six were also found in groundwater along with diethanolamines and alkyl amido betaines, which were not found in the eight archived AFFFs. This indicates that although aerobically biodegradable, hydrocarbon surfactants likely persist in groundwater due to anaerobic aquifer conditions. To the best of our knowledge, this is the first screening for hydrocarbon surfactants in AFFFs and in AFFF-contaminated groundwater.

Die Elimination von Mikroverunreinigungen in der Abwasserreinigung mit Aktivkohle oder Ozon ist eine wichtige Massnahme zum vorbeugenden Schutz der Gewässer und der Trinkwasserressourcen der Schweiz. Während der Einsatz von Ozon mit einem hohen Energieverbrauch einhergeht, führt der Einsatz von Aktivkohle andernorts zu negativen Umweltauswirkungen, da die Aktivkohle zum Teil aus fossilen Ausgangsstoffen wie Stein- oder Braunkohle hergestellt und zudem über weite Strecken transportiert wird. Aktivkohlen mit sehr guten Eliminationsleistungen könnten aber auch in der Schweiz aus einheimischen erneuerbaren Roh- und Reststoffen hergestellt werden.

RNA-based biomarkers have been successfully detected at field sites undergoing in situ bioremediation, but the detection of expressed enzymes is a more direct way to prove activity for a particular biocatalytic process of interest since they provide evidence of potential in situ activity rather than simply confirming presence and abundance of genes in a given population by measurement of DNA copies using qPCR. Here we successfully applied shotgun proteomics to field samples from a trichloroethene (TCE)-contaminated industrial site in southern Ontario, Canada that had been bio-augmented with the commercially available KB-1^TM microbial culture. The KB-1^TM culture contains multiple strains of Dehalococcoides mccartyi (D. mccartyi) as well as an organohalide respiring Geobacter species. The relative abundances of specific enzymatic proteins were subsequently compared to corresponding qPCR-derived levels of DNA and RNA biomarkers in the same samples. Samples were obtained from two wells with high hydraulic connectivity to the KB-1^TM-bioaugemented enhanced in situ bioremediation system, and two control wells that showed evidence of low levels of native organohalide respiring bacteria (OHRB), Dehalococcoides and Geobacter. Enzymes involved in organohalide respiration were detected in the metaproteomes of all four field samples, as were chaperonins of D. mccartyi, chemotaxis proteins, and ATPases. The most highly expressed RDase in the bioaugmentation culture (VcrA) was the most highly detected enzyme overall in the bioaugmented groundwater samples. In one background groundwater well, we found high expression of the Geobacter pceA RDase. The DNA and RNA biomarkers detected using qPCR-based assays were a set of orthologs of Dehalococcoides reductive dehalogenases (VcrA, TceA, BvcA, dehalogenase “DET1545”), and the Ni-Fe uptake hydrogenase, HupL. Within a sample, RNA levels for key enzymes correlated with relative protein abundance. These results indicate that laboratory observations of TCE-bioremediation biomarker protein expression are recapitulated in field environmental systems and that both RNA and protein biomarker monitoring hold promise for activity monitoring of in situ populations of OHRB.

Exposure science, in its broadest sense, studies the interactions between stressors (chemical, biological, and physical agents) and receptors (e.g. humans and other living organisms, and non-living items like buildings), together with the associated pathways and processes potentially leading to negative effects on human health and the environment. The aquatic environment may contain thousands of compounds, many of them still unknown, that can pose a risk to ecosystems and human health. Due to the unquestionable importance of the aquatic environment, one of the main challenges in the field of exposure science is the comprehensive characterization and evaluation of complex environmental mixtures beyond the classical/priority contaminants to new emerging contaminants.
The role of advanced analytical chemistry to identify and quantify potential chemical risks, that might cause adverse effects to the aquatic environment, is essential. In this paper, we present the strategies and tools that analytical chemistry has nowadays, focused on chromatography hyphenated to (high-resolution) mass spectrometry because of its relevance in this field. Key issues, such as the application of effect direct analysis to reduce the complexity of the sample, the investigation of the huge number of transformation/degradation products that may be present in the aquatic environment, the analysis of urban wastewater as a source of valuable information on our lifestyle and substances we consumed and/or are exposed to, or the monitoring of drinking water, are discussed in this article. The trends and perspectives for the next few years are also highlighted, when it is expected that new developments and tools will allow a better knowledge of chemical composition in the aquatic environment. This will help regulatory authorities to protect water bodies and to advance towards improved regulations that enable practical and efficient abatements for environmental and public health protection.

Non-target screening (NTS) including suspect screening with high resolution mass spectrometry has already shown its feasibility in detecting and identifying emerging contaminants, which subsequently triggered exposure mitigating measures. NTS has a large potential for tasks such as effective evaluation of regulations for safe marketing of substances and products, prioritization of substances for monitoring programmes and assessment of environmental quality. To achieve this, a further development of NTS methodology is required, including: (i) harmonized protocols and quality requirements, (ii) infrastructures for efficient data management, data evaluation and data sharing and (iii) sufficient resources and appropriately trained personnel in the research and regulatory communities in Europe. Recommendations for achieving these three requirements are outlined in the following discussion paper. In particular, in order to facilitate compound identification it is recommended that the relevant information for interpretation of mass spectra, as well as about the compounds usage and production tonnages, should be made accessible to the scientific community (via open-access databases). For many purposes, NTS should be implemented in combination with effect-based methods to focus on toxic chemicals.

Enhancing the understanding of the fate of wastewater-derived organic micropollutants in rivers is crucial to improve risk assessment, regulatory decision making and river management. Hyporheic exchange and sediment bacterial diversity are two factors gaining increasing importance as drivers for micropollutant degradation, but are complex to study in field experiments and usually ignored in laboratory tests aimed to estimate environmental half-lives. Flume mesocosms are useful to investigate micropollutant degradation processes, bridging the gap between the field and batch experiments. However, few studies have used flumes in this context. We present a novel experimental setup using 20 recirculating flumes and a response surface model to study the influence of hyporheic exchange and sediment bacterial diversity on half-lives of the anti-epileptic drug carbamazepine (CBZ) and the artificial sweetener acesulfame (ACS). The effect of bedform-induced hyporheic exchange was tested by three treatment levels differing in number of bedforms (0, 3 and 6). Three levels of sediment bacterial diversity were obtained by diluting sediment from the River Erpe in Berlin, Germany, with sand (1 : 10, 1 : 1000 and 1 : 100 000). Our results show that ACS half-lives were significantly influenced by sediment dilution and number of bedforms. Half-lives of CBZ were higher than ACS, and were significantly affected only by the sediment dilution variable, and thus by bacterial diversity. Our results show that (1) the flume-setup is a useful tool to study the fate of micropollutants in rivers, and that (2) higher hyporheic exchange and bacterial diversity in the sediment can increase the degradation of micropollutants in rivers.

Cyanobacterial bloom events that produce natural toxins occur in freshwaters across the globe, yet the potential risk of many cyanobacterial metabolites remains mostly unknown. Only microcystins, one class of cyanopeptides, have been studied intensively and the wealth of evidence regarding exposure concentrations and toxicity led to their inclusion in risk management frameworks for water quality. However, cyanobacteria produce an incredible diversity of hundreds of cyanopeptides beyond the class of microcystins. The question arises, whether the other cyanopeptides are in fact of no human and ecological concern or whether these compounds merely received (too) little attention thus far. Current observations suggest that an assessment of their (eco)toxicological risk is indeed relevant: First, other cyanopeptides, including cyanopeptolins and anabaenopeptins, can occur just as frequently and at similar nanomolar concentrations as microcystins in surface waters. Second, cyanopeptolins, anabaenopeptins, aeruginosins and microginins inhibit proteases in the nanomolar range, in contrast to protein phosphatase inhibition by microcystins. Cyanopeptolins, aeruginosins, and aerucyclamide also show toxicity against grazers in the micromolar range comparable to microcystins. The key challenge for a comprehensive risk assessment of cyanopeptides remains their large structural diversity, lack of reference standards, and high analytical requirements for identification and quantification. One way forward would be a prevalence study to identify the priority candidates of tentatively abundant, persistent, and toxic cyanopeptides to make comprehensive risk assessments more manageable.

The aim of the present study was to develop (i) a technique for identifying metabolites of organic contaminants by using an in vitro system of trout S9 and liquid chromatography-high-resolution mass spectrometry-based identification method and (ii) to apply this technique to identify the interactive potential of carbamazepine on the formation rate of other metabolites. The pharmaceuticals carbamazepine and propranolol and the pesticides azoxystrobin, diazinon, and fipronil were selected as test contaminants. As a result, a total of ten metabolites were identified for the five parent substances, six of which were confirmed using reference standards. Metabolic reactions included hydroxylation, epoxidation, S-oxidation, and dealkylation. The metabolic transformation rate ranged from 0.2 to 3.5 pmol/mg protein/min/μmol substrate. In the binary exposure experiment with increasing carbamazepine concentration, the formation rates of diazinon and fipronil metabolites (MDI2 and MFP2, respectively) increased, while formation of metabolites of propranolol and azoxystrobin (MPR1, MPR2, MPR3, and MAZ1) slowed down. Meanwhile, S9 pre-exposed to carbamazepine produced diazoxon, a toxic metabolite of diazinon, and pyrimidinol, a less toxic metabolite, more rapidly. These results suggest that carbamazepine, a perennial environmental pollutant, might modulate the toxicity of other substances such as diazinon but further in vivo studies are needed.

Ninety-eight soil samples were collected from farmland soils from Beijing-Tianjin-Hebei core area, Northern China, where agricultural lands were subjected to contamination from intense urban and industrial activities. Twelve organophosphates flame retardants (OPFRs) were analyzed with total soil concentrations ranging from 0.543 μg/kg to 54.9 μg/kg. Chlorinated OPFRs were dominating at mean level of 3.64 μg/kg and Tris(2-chloroisopropyl) phosphate contributed the most (mean 3.36 ± 5.61 μg/kg, 98.0%). Tris(2-ethylhexyl) phosphate was fully detected at levels of 0.041–1.95 μg/kg. Generally, tris(2-butoxyethyl) phosphate and triphenyl phosphate contributed the most to alkyl- (53.6%) and aryl-OPFRs (54.3%), respectively. The levels of ∑OPFRs close to the core urban areas were significantly higher than those from background sites. The occurrence and fate of OPFRs in soil were significantly associated with total organic carbon content and mostly with fine soil particles (<0.005 mm), and a transfer potential from the atmosphere was predicted with logK_SA values. Comparable soil levels with poly brominated diphenyl ethers s in other studies suggested that the contamination of OPFRs occurred in farmland soil with an increasing trend but currently showed no significant environmental risk based on risk quotient estimation (<1). This investigation warrants further study on behaviors of OPFRs in a soil system and a continual monitoring for their risk assessment.

Wastewater treatment plants (WWTPs) are implicated as hotspots for the dissemination of antibacterial resistance into the environment. However, the in situ processes governing removal, persistence, and evolution of resistance genes during wastewater treatment remain poorly understood. Here, we used quantitative metagenomic and metatranscriptomic approaches to achieve a broad-spectrum view of the flow and expression of genes related to antibacterial resistance to over 20 classes of antibiotics, 65 biocides, and 22 metals. All compartments of 12 WWTPs share persistent resistance genes with detectable transcriptional activities that were comparatively higher in the secondary effluent, where mobility genes also show higher relative abundance and expression ratios. The richness and abundance of resistance genes vary greatly across metagenomes from different treatment compartments, and their relative and absolute abundances correlate with bacterial community composition and biomass concentration. No strong drivers of resistome composition could be identified among the chemical stressors analyzed, although the sub-inhibitory concentration (hundreds of ng/L) of macrolide antibiotics in wastewater correlates with macrolide and vancomycin resistance genes. Contig-based analysis shows considerable co-localization between resistance and mobility genes and implies a history of substantial horizontal resistance transfer involving human bacterial pathogens. Based on these findings, we propose future inclusion of mobility incidence (M%) and host pathogenicity of antibiotic resistance genes in their quantitative health risk ranking models with an ultimate goal to assess the biological significance of wastewater resistomes with regard to disease control in humans or domestic livestock.

Groundwater is a major drinking water resource, but its quality is threatened by a broad variety of anthropogenic micropollutants (MPs), originating from agriculture, industry, or households, and undergoing various transformation processes during subsurface passage. To determine a worst-case impact of pesticide application in agriculture on groundwater quality, a target and suspect screening for more than 300 pesticides and more than 1100 pesticide transformation products (TPs) was performed in 31 Swiss groundwater samples which predominantly originated from areas with intensive agriculture. To assess additional urban contamination sources, more than 250 common urban MPs were quantified. Most of the screened pesticide TPs were experimentally observed by the pesticide producers within the European pesticide registration. To cover very polar pesticide TPs, vacuum-assisted evaporative concentration was used for enrichment, followed by liquid chromatography high-resolution tandem mass spectrometry (LC-HRMS/MS). Based on intensity, isotope pattern, retention time, and in silico fragmentation, the suspect hits were prioritised and verified. We identified 22 suspects unequivocally and five tentatively; 13 TPs are reported here for the first time to be detected in groundwater. In 13 out of 31 groundwater samples, the total concentration of the 20 identified and quantified suspects (1 pesticide and 19 pesticide TPs) exceeded the total concentration of the 519 targets (236 pesticides and TPs; 283 urban MPs) for which we screened. Pesticide TPs had higher concentrations than the parent pesticides, illustrating their importance for groundwater quality. The newly identified very polar chlorothalonil TP R471811 was the only compound detected in all samples with concentrations ranging from 3 to 2700 ng/L. Agricultural MP concentration and detection frequency correlated with agricultural land use in the catchment, except for aquifers, where protective top layers reduced MP transport from the surface. In contrast to agricultural MPs, urban MPs displayed almost no correlation with land use. The dominating entry pathway of urban MPs was river bank filtration.

Mit Screening-Methoden wurden ausgewählte Grundwasser-Proben im Rahmen einer Pilotstudie der Nationalen Grundwasserbeobachtung NAQUA auf mehrere hundert Wirkstoffe und Metaboliten von Pflanzenschutzmitteln (PSM) sowie weitere Mikroverunreinigungen untersucht. Es galt zu prüfen, ob und welche «neuen» PSM-Wirkstoffe oder -Metaboliten die Grundwasserqualität beeinträchtigen und in das NAQUA-Langzeitmonitoring integriert werden sollten. Mehr als 20 PSM-Metaboliten wurden erstmals im Grundwasser nachgewiesen. Diejenigen des Fungizids Chlorthalonil waren aufgrund hoher Konzentrationen und weiter Verbreitung besonders auffällig.

Wastewater treatment plant effluents are important point sources of micropollutants. To assess how the discharge of treated wastewater affects the ecotoxicity of small to medium-sized streams we collected water samples up- and downstream of 24 wastewater treatment plants across the Swiss Plateau and the Jura regions of Switzerland. We investigated estrogenicity, inhibition of algal photosynthetic activity (photosystem II, PSII) and growth, and acetylcholinesterase (AChE) inhibition. At four sites, we measured feeding activity of amphipods (Gammarus fossarum) in situ as well as water flea (Ceriodaphnia dubia) reproduction in water samples. Ecotoxicological endpoints were compared with results from analyses of general water quality parameters as well as a target screening of a wide range of organic micropollutants with a focus on pesticides and pharmaceuticals using liquid chromatography high-resolution tandem mass spectrometry. Measured ecotoxicological effects in stream water varied substantially among sites: 17β-estradiol equivalent concentrations (EEQ_bio, indicating the degree of estrogenicity) were relatively low and ranged from 0.04 to 0.85 ng/L, never exceeding a proposed effect-based trigger (EBT) value of 0.88 ng/L. Diuron equivalent (DEQ_bio) concentrations (indicating the degree of photosystem II inhibition in algae) ranged from 2.4 to 1576 ng/L and exceeded the EBT value (70 ng/L) in one third of the rivers studied, sometimes even upstream of the WWTP. Parathion equivalent (PtEQ_bio) concentrations (indicating the degree of AChE inhibition) reached relatively high values (37 to 1278 ng/L) mostly exceeding the corresponding EBT (196 ng/L PtEQ_bio). Decreased feeding activity by amphipods or decreased water flea reproduction downstream compared to the upstream site was observed at one of four investigated sites only. Results of the combined algae assay (PSII inhibition) correlated best with results of chemical analysis for PSII inhibiting herbicides. Estrogenicity was partly and AChE inhibition strongly underestimated based on measured steroidal estrogens respectively organophosphate and carbamate insecticides. An impact of dissolved organic carbon on results of the AChE inhibition assay was obvious. For this assay more work is required to further explore the missing correlation of bioassay data with chemical analytical data. Overall, the discharge of WWTP effluent led to increased estrogenicity, PSII and AChE inhibition downstream, irrespective of upstream land use.

Soil erosion and associated sediment transfer are among the major causes of aquatic ecosystem and surface water quality impairment. Through land use and agricultural practices, human activities modify the soil erosive risk and the catchment connectivity, becoming a key factor of sediment dynamics. Hence, restoration and management plans of water bodies can only be efficient if the sediment sources and the proportion attributable to different land uses are identified. According to this aim, we applied two approaches, namely compound-specific isotope analysis (CSIA) of long-chain fatty acids (FAs) and triterpenoid biomarker analysis, to a eutrophic lake, Lake Baldegg, and its agriculturally used catchment (Switzerland). Soils reflecting the five main land uses of the catchment (arable lands, temporary and permanent grasslands, mixed forests, orchards) were subjected to CSIA. The compound-specific stable isotope δ¹³C signatures clearly discriminate between potential grasslands (permanent and temporary) and forest sources. Signatures of agricultural land and orchards fall in between. The soil signal was compared to the isotopic signature of a lake sediment sequence covering ca. 130 years (before 1885 to 2009). The recent lake samples (1940 to 2009, with the exception of 1964 to 1972) fall into the soil isotopic signature polygon and indicate an important contribution of the forests, which might be explained by (1) the location of the forests on steep slopes, resulting in a higher connectivity of the forests to the lake, and/or (2) potential direct inputs of trees and shrubs growing along the rivers feeding the lake and around the lake. However, the lake sediment samples older than 1940 lie outside the source soils' polygon, as a result of FA contribution from a not yet identified source, most likely produced by an in situ aquatic source, either algae, bacteria or other microorganisms or an ex-site historic source from wetland soils and plants (e.g. Sphagnum species). Despite the overprint of the yet unknown source on the historic isotopic signal of the lake sediments, land use and catchment history are clearly reflected in the CSIA results, with isotopic shifts being synchronous with changes in the catchment, land use and eutrophication history. The investigated highly specific biomarkers were not detected in the lake sediment, even though they were present in the soils. However, two trimethyltetrahydrochrysenes (TTHCs), natural diagenetic products of pentacyclic triterpenoids, were found in the lake sediments. Their origin is attributed to the in situ microbial degradation of some of the triterpenoids. While the need to apportion sediment sources is especially crucial in eutrophic systems, our study stresses the importance of exercising caution with CSIA and triterpenoid biomarkers in such environments, where the active metabolism of bacteria might mask the original terrestrial isotopic signals.

Rivers are important ecosystems under continuous anthropogenic stresses. The hyporheic zone is a ubiquitous, reactive interface between the main channel and its surrounding sediments along the river network. We elaborate on the main physical, biological, and biogeochemical drivers and processes within the hyporheic zone that have been studied by multiple scientific disciplines for almost half a century. These previous efforts have shown that the hyporheic zone is a modulator for most metabolic stream processes and serves as a refuge and habitat for a diverse range of aquatic organisms. It also exerts a major control on river water quality by increasing the contact time with reactive environments, which in turn results in retention and transformation of nutrients, trace organic compounds, fine suspended particles, and microplastics, among others. The paper showcases the critical importance of hyporheic zones, both from a scientific and an applied perspective, and their role in ecosystem services to answer the question of the manuscript title. It identifies major research gaps in our understanding of hyporheic processes. In conclusion, we highlight the potential of hyporheic restoration to efficiently manage and reactivate ecosystem functions and services in river corridors.

Methylorubrum extorquens (formerly Methylobacterium extorquens) AM1 is a methylotrophic bacterium with a versatile lifestyle. Various carbon sources including acetate, succinate and methanol are utilized by M. extorquens AM1 with the latter being a promising inexpensive substrate for use in the biotechnology industry. Itaconic acid (ITA) is a high-value building block widely used in various industries. Given that no wildtype methylotrophic bacteria are able to utilize methanol to produce ITA, we tested the potential of M. extorquens AM1 as an engineered host for this purpose. In this study, we successfully engineered M. extorquens AM1 to express a heterologous codon-optimized gene encoding cis-aconitic acid decarboxylase. The engineered strain produced ITA using acetate, succinate and methanol as the carbon feedstock. The highest ITA titer in batch culture with methanol as the carbon source was 31.6 ± 5.5 mg/L, while the titer and productivity were 5.4 ± 0.2 mg/L and 0.056 ± 0.002 mg/L/h, respectively, in a scaled-up fed-batch bioreactor under 60% dissolved oxygen saturation. We attempted to enhance the carbon flux toward ITA production by impeding poly-β-hydroxybutyrate accumulation, which is used as carbon and energy storage, via mutation of the regulator gene phaR. Unexpectedly, ITA production by the phaR mutant strain was not higher even though poly-β-hydroxybutyrate concentration was lower. Genome-wide transcriptomic analysis revealed that phaR mutation in the ITA-producing strain led to complex rewiring of gene transcription, which might result in a reduced carbon flux toward ITA production. Besides poly-β-hydroxybutyrate metabolism, we found evidence that PhaR might regulate the transcription of many other genes including those encoding other regulatory proteins, methanol dehydrogenases, formate dehydrogenases, malate:quinone oxidoreductase, and those synthesizing pyrroloquinoline quinone and thiamine co-factors. Overall, M. extorquens AM1 was successfully engineered to produce ITA using acetate, succinate and methanol as feedstock, further supporting this bacterium as a feasible host for use in the biotechnology industry. This study showed that PhaR could have a broader regulatory role than previously anticipated, and increased our knowledge of this regulator and its influence on the physiology of M. extorquens AM1.

Aliphatic amines are common constituents in micropollutants and dissolved organic matter and present in elevated concentrations in wastewater-impacted source waters. Due to high reactivity, reactions of aliphatic amines with ozone are likely to occur during ozonation in water and wastewater treatment. We investigated the kinetics and mechanisms of the reactions of ozone with ethylamine, diethylamine, and triethylamine as model nitrogenous compounds. Species-specific second-order rate constants for the neutral parent amines ranged from 9.3 × 10⁴ to 2.2 × 10⁶ M⁻¹s⁻¹ and the apparent second-order rate constants at pH 7 for potential or identified transformation products were 6.8 × 10⁵ M⁻¹s⁻¹ for N,N-diethylhydroxylamine, ∼10⁵ M⁻¹s⁻¹ for N-ethylhydroxylamine, 1.9 × 10³ M⁻¹s⁻¹ for N-ethylethanimine oxide, and 3.4 M⁻¹s⁻¹ for nitroethane. Product analyses revealed that all amines were transformed to products containing a nitrogen-oxygen bond (e.g., triethylamine N-oxide and nitroethane) with high yields, i.e., 64–100% with regard to the abated target amines. These findings could be confirmed by measurements of singlet oxygen and hydroxyl radical which are formed during the amine-ozone reactions. Based on the high yields of nitroethane from ethylamine and diethylamine, a significant formation of nitroalkanes can be expected during ozonation of waters containing high levels of dissolved organic nitrogen, as expected in wastewaters or wastewater-impaired source waters. This may pose adverse effects on the aquatic environment and human health.

For the study of different levels of (intra)cellular regulation and condition-dependent insight into metabolic activities, fluxomics experiments based on stable isotope tracer experiments using ¹³C have become a well-established approach. The experimentally obtained non-naturally distributed ¹³C labeling patterns of metabolite pools can be measured by mass spectrometric detection with front-end separation and can be consequently incorporated into biochemical network models. Here, despite a tedious derivatization step, gas chromatographic separation of polar metabolites is favorable because of the wide coverage range and high isomer separation efficiency. However, the typically employed electron ionization energy of 70 eV leads to significant fragmentation and consequently only low-abundant ions with an intact carbon backbone. Since these ions are considered a prerequisite for the analysis of the non-naturally distributed labeling patterns and further integration into modeling strategies, a softer ionization technique is needed. In the present work, a novel low energy electron ionization source is optimized for the analysis of primary metabolites and compared with a chemical ionization approach in terms of trueness, precision, and sensitivity.

Rationale: The wide chemical diversity and complex matrices inherent to metabolomics still pose a challenge to current analytical approaches for metabolite screening. Although dedicated front‐end separation techniques combined with high‐resolution mass spectrometry set the benchmark from an analytical point of view, the increasing number of samples and sample complexity demand for a compromise in terms of selectivity, sensitivity and high‐throughput analyses.
Methods: Prior to low‐field drift tube ion mobility (IM) separation and quadrupole time‐of‐flight mass spectrometry (QTOFMS) detection, rapid ultrahigh‐performance liquid chromatography separation was used for analysis of different concentration levels of dansylated metabolites present in a yeast cell extract. For identity confirmation of metabolites at the MS2 level, an alternating frame approach was chosen and two different strategies were tested: a data‐independent all‐ions acquisition and a quadrupole broad band isolation (Q‐BBI) directed by IM drift separation.
Results: For Q‐BBI analysis, the broad mass range isolation was successfully optimized in accordance with the distinctive drift time to m/z correlation of the dansyl derivatives. To guarantee comprehensive sampling, a broad mass isolation window of 70 Da was employed. Fragmentation was performed via collision‐induced dissociation, applying a collision energy ramp optimized for the dansyl derivatives. Both approaches were studied in terms of linear dynamic range and repeatability employing ethanolic extracts of Pichia pastoris spiked with 1 μM metabolite mixture. Example data obtained for histidine and glycine showed that drift time precision (<0.01 to 0.3% RSD, n = 5) compared very well with the data reported in an earlier IM‐TOFMS‐based study.
Conclusions: Chimeric mass spectra, inherent to data‐independent analysis approaches, are reduced when using a drift time directed Q‐BBI approach. Additionally, an improved linear dynamic working range was observed, representing, together with a rapid front‐end separation, a powerful approach for metabolite screening.

The mutagenicity of four organic UV filters namely oxybenzone (benzophenone-3), dioxybenzone (benzophenone-8), avobenzone, and octyl methoxycinnamate, in chlorinated bromide-rich water (artificial seawater) was investigated. Mutagenicity was evaluated using Ames test in Salmonella typhimurium TA98 without S9 mix. Chemical analysis using high-resolution mass spectrometry was carried out to elucidate the mutagenic transformation products. Among the studied UV filters, only dioxybenzone exhibited a clear mutagenic activity following chlorination in seawater at ratio 1:10 (UV filter:chlorine). In contrast, no mutagenic activity was detected when chlorine was added at higher doses (ratio 1:1000). High-resolution mass spectrometry analysis showed that mutagenic extracts contained several brominated transformation products of dioxybenzone. Time course analysis of the transformation products at increasing chlorine doses showed that they were unstable and disappeared more quickly at higher chlorine doses. This instability explained the absence of mutagenic activity of dioxybenzone when 1000-fold excess chlorine was added, as no transformation products were detected. Relevance of these findings to the context of swimming pool is discussed. Further investigations taking into consideration the mutagenicity of not only the intermediate transformation products but also the final disinfection byproducts are needed to determine the overall impact of high levels of chlorine on the overall mutagenicity. This study highlights the importance of considering the reactivity of organic UV filters and their transformation products in disinfected recreational waters when sunscreen formulations are prepared.

A remaining challenge within microbial ecology is to understand the determinants of richness and diversity observed in environmental microbial communities. In a range of systems, including activated sludge bioreactors, the microbial residence time (MRT) has been previously shown to shape the microbial community composition. However, the physiological and ecological mechanisms driving this influence have remained unclear. Here, this relationship is explored by analyzing an activated sludge system fed with municipal wastewater. Using a model designed in this study based on Monod-growth kinetics, longer MRTs were shown to increase the range of growth parameters that enable persistence, resulting in increased richness and diversity in the modeled community. In laboratory experiments, six sequencing batch reactors treating domestic wastewater were operated in parallel at MRTs between 1 and 15 days. The communities were characterized using both 16S ribosomal RNA and non-target messenger RNA sequencing (metatranscriptomic analysis), and model-predicted monotonic increases in richness were confirmed in both profiles. Accordingly, taxonomic Shannon diversity also increased with MRT. In contrast, the diversity in enzyme class annotations resulting from the metatranscriptomic analysis displayed a non-monotonic trend over the MRT gradient. Disproportionately high abundances of transcripts encoding for rarer enzymes occur at longer MRTs and lead to the disconnect between taxonomic and functional diversity profiles.

Passive sampling is a well-established tool for monitoring time-weighted average concentrations of polar and semi-polar organic contaminants in streams at flow velocities between 0.1 and 0.4 m s⁻¹. However, its application under low-flow conditions (10⁻⁵ to 0.01 m s⁻¹) – as encountered in hyporheic zones – has been scarcely reported. In this study, 3 novel passive sampler configurations were developed for the monitoring of (semi-)polar organic pollutants and related transformation products across the water-sediment interface and thus across varying hydrodynamic conditions. Their design was inspired by Chemcatcher and diffusive gradients in thin films for organics. To determine the most optimal sampler design, an uptake experiment was completed involving the 3 novel passive sampler configurations and a reference Chemcatcher in polar configuration. The experiments consisted of a circular flume that simulated the main channel of a stream and an aquarium with stagnant water that represented the underlying hyporheic zone. The systems were exposed to 192 organic pollutants at environmental concentrations, and the samplers were then collected, extracted and analyzed using liquid chromatography high-resolution mass spectrometry after 2, 6 and 14 days. The configuration that was most insensitive to different hydrodynamic conditions consisted of a reversed-phase sulfonated styrenedivinylbenzene disk as the receiving phase that was covered by an agarose diffusion gel and topped with a polyethersulfone membrane filter. To further evaluate its environmental application, samplers were installed downstream of a sewage treatment plant located at an urban stream in Berlin, Germany (Erpe). The samplers were mounted on custom-made holders which were subsequently embedded in the stream bed to position samplers above (0.30 m) and within the sediment (−0.15/-0.30/-0.45 m) for 11 days. Target and suspect screening workflows were then applied to identify common concentration patterns and link parent attenuation to transformation product formation. A total of 104 concentration profiles were determined, suggesting the efficiency of the proposed sampling strategy in the water-sediment interface. Valsartan acid was the only known transformation product indicative of hyporheic zone-driven attenuation as its concentration in porewater by far exceeded its concentration in surface water. Similar patterns were observed for a larger list of suspected transformation products, of which a sotalol transformation product was tentatively identified. Overall, the established sampling methodology can be effectively used to quantify organic contaminants during low-flow conditions and is suitable for the characterization of attenuation patterns of organic pollutants in hyporheic zones.

Every day, manifold synthetic organic chemicals enter the water cycle via sewage systems and end up in surface waters via discharge from wastewater treatment plants. Advances in analytical instrumentation led to the discovery of pollutants of emerging concern (e.g. pharmaceuticals, personal care products, illicit drugs), many of which are inherently polar and mobile in the aquatic environment. Since their removal in wastewater and water treatment is often incomplete, these compounds, but also their transformation products, have a significant potential to accumulate in the water cycle, especially if they are persistent and widely emitted. Elucidation of their environmental fate and the development of suitable analytical methods are therefore of critical importance. Fate studies have proven an essential role of sediments in the elimination of organic pollutants in lotic waters. In particular, the biodegradation of polar organic pollutants in the water-saturated sediments below the stream bed and in the adjacent stream banks, collectively referred to as hyporheic zones, is assumed to contribute significantly to the self-purification of natural waters. [...]

Täglich gelangen unterschiedlichste synthetische organische Chemikalien durch Abwassersysteme in den Wasserkreislauf und über das behandelte Abwasser von Kläranlagen in Oberflächengewässer. Fortschritte in der instrumentellen Analytik führten zur Entdeckung neuer umweltrelevanter Schadstoffklassen, sogenannter «emerging contaminants» (z. B. Pharmazeutika und Körperpflegeprodukte), die oftmals polar und dadurch mobil in der aquatischen Umwelt sind. In der Trinkwasseraufbereitung und Abwasserbehandlung werden sie häufig nur unvollständig eliminiert, und haben dadurch ein erhöhtes Potential, sich im Wasserkreislauf, zusammen mit ihren Transformationsprodukten, anzureichern. Dies gilt insbesondere, wenn sie nicht nur polar, sondern auch persistent sind und vielerorts emittiert werden. Aus diesen Gründen ist es wichtig, den Verbleib dieser Stoffe in der Umwelt zu untersuchen, und die hierfür notwendigen instrumentellen Analysemethoden zu entwickeln. Umweltstudien konnten eine wesentliche Rolle von Sedimenten beim Abbau organischer Schadstoffe in Fliessgewässern aufweisen. Besonders der biologische Abbau polarer organischer Schadstoffe in wassergesättigten Sedimenten unterhalb des Flussbetts und in der angrenzenden Flussbank, den sogenannten «hyporheischen Zonen», sollen zur Selbstreinigung natürlicher Gewässer beitragen. In der vorliegenden Doktorarbeit wurden Vorkommen, Verbleib und biologischer Abbau einer Vielzahl organischer Schadstoffe in hyporheischen Zonen untersucht. Hierfür wurden unterschiedliche Ansätze verfolgt. Zum einen wurden organische Schadstoffe vom Oberflächenwasser eines urbanen Flusses in das darunterliegende Sediment verfolgt, zum anderen wurden die Einflüsse von hyporheischem Austausch und bakterieller Diversität auf den Abbau von Schadstoffen in rezirkulierenden Wasser-Sediment Rinnensystemen untersucht. In diesem Zusammenhang wurden eine geeignete Methode zur passiven Probenahme und zwei neue Analysemethoden entwickelt, validiert und teilweise im Feld oder in den Rinnensystemen angewandt. [...]

Vacuum-assisted evaporative concentration (VEC) was successfully applied and validated for the enrichment of 590 organic substances from river water and wastewater. Different volumes of water samples (6 mL wastewater influent, 15 mL wastewater effluent, and 60 mL river water) were evaporated to 0.3 mL and finally adjusted to 0.4 mL. 0.1 mL of the concentrate were injected into a polar reversed-phase C18 liquid chromatography column coupled with electrospray ionization to high-resolution tandem mass spectrometry. Analyte recoveries were determined for VEC and compared against a mixed-bed multilayer solid-phase extraction (SPE). Both approaches performed equally well (≥ 70% recovery) for a vast number of analytes (n = 327), whereas certain substances were especially amenable to enrichment by either SPE (e.g., 4-chlorobenzophenone, logD_ow,pH7 4) or VEC (e.g., TRIS, logD_ow,pH7 − 4.6). Overall, VEC was more suitable for the enrichment of polar analytes, albeit considerable signal suppression (up to 74% in river water) was observed for the VEC-enriched sample matrix. Nevertheless, VEC allowed for accurate and precise quantification down to the sub-nanogram per liter level and required no more than 60 mL of the sample, as demonstrated by its application to several environmental water matrices. By contrast, SPE is typically constrained by high sample volumes ranging from 100 mL (wastewater influent) to 1000 mL (river water). The developed VEC workflow not only requires low labor cost and minimum supervision but is also a rapid, convenient, and environmentally safe alternative to SPE and highly suitable for target and non-target analysis.

The widespread application of herbicides impacts surface water and groundwater. Metabolites (e.g., desphenylchloridazon from chloridazon) may be persistent and even more polar than the parent herbicide, which increases the risk of groundwater contamination. When parent herbicides are still applied, metabolites are constantly formed and may also be degraded. Evaluating their degradation on the basis of concentration measurements is, therefore, difficult. This study presents compound-specific stable-isotope analysis (CSIA) of nitrogen- and carbon-isotope ratios at natural abundances as an alternative analytical approach to track the origin, formation, and degradation of desphenylchloridazon (DPC), the major degradation product of the herbicide chloridazon. Methods were developed and validated for carbon- and nitrogen-isotope analysis (δ¹³C and δ¹⁵N) of DPC by liquid chromatography–isotope-ratio mass spectrometry (LC-IRMS) and derivatization gas chromatography–IRMS (GC-IRMS), respectively. Injecting standards directly onto an Atlantis LC-column resulted in reproducible δ¹³C-isotope analysis (standard deviation <0.5‰) by LC-IRMS with a limit of precise analysis of 996 ng of DPC on-column. Accurate and reproducible δ¹⁵N analysis with a standard deviation of <0.4‰ was achieved by GC-IRMS after derivatization of >100 ng of DPC with 160-fold excess of (trimethylsilyl)diazomethane. Application of the method to environmental-seepage water indicated that newly formed DPC could be distinguished from "old" DPC by the different isotopic signatures of the two DPC sources.

Aufgrund der Befunde von Pyrethroiden und Organophosphaten in Oberflächengewässern aus den Pilotstudien NAWA SPEZ 2012 und 2015 haben sich Ende 2016 mehrere Kantonale Gewässerschutzlabore mit der Eawag und der VSA-Plattform "Gewässerqualität" zu einer Arbeitsgruppe zusammengeschlossen. Ziel der Task-Force "Pyrethroide" war, Pyrethroide und Organophosphate routinemässig in Fliessgewässern bis in den tiefen Picogramm-pro-Liter-Bereich zu messen. Gut drei Jahre später sind die Erfolge dieser Arbeitsgruppe sichtbar.

Niederschlags- und Mischabwasserentlastungen aus Siedlungen tragen zu Mikroverunreinigungen in Gewässern bei. Mit Passivsammlern wurden erstmals Konzentrationsbereiche an 20 Standorten für insgesamt 95 Ereignisse bestimmt, was die Analyse von Unterschieden zwischen Standorten sowie die Identifikation relevanter Eintragspfade unterstützt. An 13 Standorten waren bei mindestens einer Substanz die Konzentrationen im Mischabwasser höher als akute Qualitätskriterien.

Contaminants in sewer overflows can contribute to exceedances of environmental quality standards, thus the quantification of contaminants during rainfall events is of relevance. However, monitoring is challenged by i) high spatiotemporal variability of contaminants in events of hard-to-predict durations, and ii) a large number of remote sites, which would imply enormous efforts with traditional sampling equipment. Therefore, we evaluate the applicability of passive samplers (Empore styrene-divinylbenzene reverse phase sulfonated (SDB-RPS)) to monitor a set of 13 polar organic contaminants. We present calibration experiments at high temporal resolution to assess the rate limiting accumulation mechanisms for short events (<36 h), report parameters for typical sewer conditions and compare passive samplers with composite water samples in a field study (three locations, total 10 events). With sampling rates of 0.35–3.5 L/d for 1 h reference time, our calibration results indicate a high sensitivity of passive samplers to sample short, highly variable sewer overflows. The contaminant uptake kinetic shows a fast initial accumulation, which is not well represented with the typical first-order model. Our results indicate that mass transfer to passive samplers is either controlled by the water boundary layer and the sorbent, or by the sorbent alone. Overall, passive sampler concentration estimates are within a factor 0.4 to 3.1 in comparison to composite water samples in the field study. We conclude that passive samplers are a promising approach to monitor a large number of discharge sites although it cannot replace traditional stormwater quality sampling in some cases (e.g. exact load estimates, high temporal resolution). Passive samplers facilitate identifying and prioritizing locations that may require more detailed investigations.

Tandem mass spectral databases are indispensable for fast and reliable compound identification in nontargeted analysis with liquid chromatography–high resolution tandem mass spectrometry (LC-HRMS/MS), which is applied to a wide range of scientific fields. While many articles now review and compare spectral libraries, in this manuscript we investigate two high-quality and specialized collections from our respective institutes, recorded on different instruments (quadrupole time-of-flight or QqTOF vs. Orbitrap). The optimal range of collision energies for spectral comparison was evaluated using 233 overlapping compounds between the two libraries, revealing that spectra in the range of CE 20–50 eV on the QqTOF and 30–60 nominal collision energy units on the Orbitrap provided optimal matching results for these libraries. Applications to complex samples from the respective institutes revealed that the libraries, combined with a simple data mining approach to retrieve all spectra with precursor and fragment information, could confirm many validated target identifications and yield several new Level 2a (spectral match) identifications. While the results presented are not surprising in many ways, this article adds new results to the debate on the comparability of Orbitrap and QqTOF data and the application of spectral libraries to yield rapid and high-confidence tentative identifications in complex human and environmental samples.