Incorporation of sediment- and soil-specific aspects in the Criteria for Reporting and Evaluating Ecotoxicity Data (CRED).

In environmental risk assessment either for registration purposes or for retrospective assessments of monitoring data, the hazard assessment is predominantly based on effect data from ecotoxicity studies. Most regulatory frameworks require studies used for risk assessment to be evaluated for reliability and relevance. Historically, the Klimisch methodology was used in many regulatory procedures where reliability needed to be evaluated. More recently, the Criteria for Reporting and Evaluating Ecotoxicity Data (CRED) have been developed for aquatic ecotoxicity studies, providing more detailed guidance on the evaluation and reporting of not only the reliability but also the relevance of a scientific study. Here, we discuss the application of the CRED methodology for assessing sediment and soil ecotoxicity studies, addressing important sediment- and soil-specific criteria that should be included as part of the CRED evaluation system. We also provide detailed recommendations for the design and reporting of sediment and soil toxicity studies that can be used by scientists and researchers wishing to contribute ecotoxicological data for effect assessments carried out within regulatory frameworks. Integr Environ Assess Manag 2024;00:1-13. © 2024 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Mountain Hiking: Prolonged Eccentric Muscle Contraction during Simulated Downhill Walking Perturbs Sensorimotor Control Loops Needed for Safe Dynamic Foot-Ground Interactions.

Safe mountain hiking requires precise control of dynamic foot-ground interactions. In addition to vision and vestibular afferents, limb proprioception, sensorimotor control loops, and reflex responses are used to adapt to the specific nature of the ground contact. Diminished leg dexterity and balance during downhill walking is usually attributed to fatigue. We investigated the supplementary hypothesis that the eccentric contractions inherent to downhill walking can also disrupt muscle proprioception, as well as the sensorimotor control loops and reflex responses that depend on it. In this study, we measured leg dexterity (LD), anterior-posterior (AP) and medio-lateral (ML) bipedal balance, and maximal voluntary leg extension strength in young and healthy participants before and after 30 min of simulated downhill walking at a natural pace on a treadmill at a 20° decline. Post-pre comparisons of LD (p < 0.001) and AP balance (p = 0.001) revealed significant reductions in dynamic foot-ground interactions after eccentric exercise without an accompanying reduction in leg extension strength. We conclude that eccentric contractions during downhill walking can disrupt the control of dynamic foot-ground interactions independently of fatigue. We speculate that mountaineering safety could be improved by increasing conscious attention to compensate for unadjusted proprioception weighting, especially in the descent.

Focus of Attention in Coach Instructions for Technique Training in Sports: A Scrutinized Review of Review Studies.

Literature reports superior performance when focusing one's attention during a movement on environmental effects of that movement (external focus, EF) compared to focusing on the moving body (internal focus, IF). Nevertheless, IF instructions still play an important role in the daily practice of coaches, trainers, and therapists. The current review compiles evidence for focus-of-attention concepts on movement form corrections and technique training. Reviews on the topic and selected additional papers addressing the effect of attentional focus on movement form or on kinetic, kinematic or muscle activity data were included. Both EF and IF instructions affect movement form. The reviews revealed that IF instructions seem to be better applicable to direct movement form changes than EF instructions. In contrast, EF instructions better facilitate optimization within the whole-body coordination, often resulting in better performance outcomes not directly linked to movement pattern changes. Several studies discuss focus-of-attention effects in the context of the optimal feedback control theory expanding on the constrained action hypothesis. In summary, EF and IF instructions both affect form and performance of movements, however, their relative efficacy is situation dependent. The often-purported superiority of EF over IF instructions cannot be generalized to all application contexts.

Whole-body movement analysis using principal component analysis: What is the internal consistency between outcomes originating from the same movement simultaneously recorded with different measurement devices?

A growing number of studies apply Principal Component Analysis (PCA) on whole-body kinematic data to facilitate an analysis of posture changes in human movement. An unanswered question is, how much the PCA outcomes depend on the chosen measurement device. This study aimed to assess the internal consistency of PCA outcomes from treadmill walking motion capture data simultaneously collected through laboratory-grade optical motion capture and field-suitable inertial-based motion tracking. Data was simultaneously collected using VICON (whole-body plug-in gait marker positions) and Xsens (body segment positions) from 20 participants during 2-min treadmill walking. Using PCA, Principal Movements (PMs) were determined using two commonly used practices: on an individual and a grouped basis. For both, correlation matrices were used to determine internal consistency between outcomes from either measurement system for each PM. Both individual and grouped approach showed excellent internal consistency between outcomes from the two systems among the lower order PMs. For the individual analysis, high correlations were only found along the diagonal of the correlation matrix while the grouped analysis also showed high off-diagonal correlations. These results have important implications for future application of PCA in terms of the independence of the resulting PM data, the way group-differences are expressed in higher-order PMs and the interpretation of movement complexity. Concluding, while PCA-outcomes from the two systems start to deviate in the higher order PMs, excellent internal consistency was found in the lower order PMs which already represent about 98% of the variance in the dataset.

Intervention for Better Knee Alignment during Jump Landing: Is There an Effect of Internally vs. Externally Focused Instructions?

Externally focused attention is known to induce superior results in the movement outcome, whereas focusing attention on the moving body (internal focus) causes conscious control and constrains action. The study investigated effects on knee trajectory and whole-body movement complexity when addressing knee alignment using externally (EF) vs. internally (IF) focused instructions. Young ski racers, n = 24 (12 male), performed landings with subsequent jumps to submaximal height. Movements were tracked and analyzed during the ground contact phase. Sets of jumps were executed without instruction (CON), followed by EF and IF instructions on knee alignment in a random order. Medial-lateral displacement of the knee in landing quantified task achievement, and whole-body principal component analysis was used to compute movement complexity. Knee alignment instructions led to a significantly lower medial knee displacement compared to CON (p = 0.001, ηp2 = 0.35). EF vs. IF did not reach significance. EF, as well as IF instructions increased the prominence of the first movement pattern (p = 0.01, ηp2 = 0.22) with a reduction of higher-order patterns (p = 0.002, W = 0.11), suggesting a strategy of freezing degrees of freedom. Both instructions addressing the movement form positively influenced knee displacement during landing, and both led to a freezing strategy, simplifying whole-body coordination.

Evaluation of a full-scale wastewater treatment plant with ozonation and different post-treatments using a broad range of in vitro and in vivo bioassays.

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.

Biological effect and chemical monitoring of Watch List substances in European surface waters: Steroidal estrogens and diclofenac - Effect-based methods for monitoring frameworks.

Three steroidal estrogens, 17α-ethinylestradiol (EE2), 17ß-estradiol (E2), estrone (E1), and the non-steroidal anti-inflammatory drug (NSAID), diclofenac have been included in the first Watch List of the Water Framework Directive (WFD, EU Directive 2000/60/EC, EU Implementing Decision 2015/495). This triggered the need for more EU-wide surface water monitoring data on these micropollutants, before they can be considered for inclusion in the list of priority substances regularly monitored in aquatic ecosystems. The revision of the priority substance list of the WFD offers the opportunity to incorporate more holistic bioanalytical approaches, such as effect-based monitoring, alongside single substance chemical monitoring. Effect-based methods (EBMs) are able to measure total biological activities (e.g., estrogenic activity or cyxlooxygenase [COX]-inhibition) of specific group of substances (such as estrogens and NSAIDs) in the aquatic environment at low concentrations (pg/L). This makes them potential tools for a cost-effective and ecotoxicologically comprehensive water quality assessment. In parallel, the use of such methods could build a bridge from chemical status assessments towards ecological status assessments by adressing mixture effects for relevant modes of action. Our study aimed to assess the suitability of implementing EBMs in the WFD, by conducting a large-scale sampling and analysis campaign of more than 70 surface waters across Europe. This resulted in the generation of high-quality chemical and effect-based monitoring data for the selected Watch List substances. Overall, water samples contained low estrogenicity (0.01-1.3 ng E2-Equivalent/L) and a range of COX-inhibition activity similar to previously reported levels (12-1600 ng Diclofenac-Equivalent/L). Comparison between effect-based and conventional analytical chemical methods showed that the chemical analytical approach for steroidal estrogens resulted in more (76%) non-quantifiable data, i.e., concentrations were below detection limits, compared to the EBMs (28%). These results demonstrate the excellent and sensitive screening capability of EBMs.

Clozapine modulation of zebrafish swimming behavior and gene expression as a case study to investigate effects of atypical drugs on aquatic organisms.

Mental illnesses affect more than 150 million people in Europe and lead to an increasing consumption of neuroactive drugs during the last twenty years. The antipsychotic compound, clozapine, is one of the most used psychotropic drugs worldwide, with potentially negative consequences for the aquatic environment. Hence, the objectives of the study presented here were the quantification of clozapine induced changes in swimming behavior of exposed Danio rerio embryos and the elucidation of the molecular effects on the serotonergic and dopaminergic systems. Yolk-sac larvae were exposed to different concentrations (0.2 mg/L, 0.4 mg/L, 0.8 mg/L, 1.6 mg/L, 3.2 mg/L and 6.4 mg/L) of clozapine for 116 h post-fertilization, and changes in the swimming behavior of the larvae were assessed. Further, quantitative real-time PCR was performed to analyze the expression of selected genes. The qualitative evaluation of changes in the swimming behavior of D. rerio larvae revealed a significant decrease of the average swimming distance and velocity in the light-dark transition test, with more than a 36% reduction at the highest exposure concentration of 6.4 mg/L. Furthermore, the total larval body length was reduced at the highest concentration. An in-depth analysis based on expression of selected target genes of the serotonin (slc6a4a) and dopamine (drd2a) system showed an upregulation at a concentration of 1.6 mg/L and above. In addition, a lower increase in expression was detected for biomarkers of general stress (adra1a and cyp1a2). Our data show that exposure to clozapine during development inhibits swimming activity of zebrafish larvae, which could, in part, be due to disruption of the serotonin- and dopamine system.

Implications of Optimal Feedback Control Theory for Sport Coaching and Motor Learning: A Systematic Review.

Best practice in skill acquisition has been informed by motor control theories. The main aim of this study is to screen existing literature on a relatively novel theory, Optimal Feedback Control Theory (OFCT), and to assess how OFCT concepts can be applied in sports and motor learning research. Based on 51 included studies with on average a high methodological quality, we found that different types of training seem to appeal to different control processes within OFCT. The minimum intervention principle (founded in OFCT) was used in many of the reviewed studies, and further investigation might lead to further improvements in sport skill acquisition. However, considering the homogenous nature of the tasks included in the reviewed studies, these ideas and their generalizability should be tested in future studies.

Estrogenicity of chemical mixtures revealed by a panel of bioassays.

Estrogenic compounds are widely released to surface waters and may cause adverse effects to sensitive aquatic species. Three hormones, estrone, 17ß-estradiol and 17α-ethinylestradiol, are of particular concern as they are bioactive at very low concentrations. Current analytical methods are not all sensitive enough for monitoring these substances in water and do not cover mixture effects. Bioassays could complement chemical analysis since they detect the overall effect of complex mixtures. Here, four chemical mixtures and two hormone mixtures were prepared and tested as reference materials together with two environmental water samples by eight laboratories employing nine in vitro and in vivo bioassays covering different steps involved in the estrogenic response. The reference materials included priority substances under the European Water Framework Directive, hormones and other emerging pollutants. Each substance in the mixture was present at its proposed safety limit concentration (EQS) in the European legislation. The in vitro bioassays detected the estrogenic effect of chemical mixtures even when 17ß-estradiol was not present but differences in responsiveness were observed. LiBERA was the most responsive, followed by LYES. The additive effect of the hormones was captured by ERα-CALUX, MELN, LYES and LiBERA. Particularly, all in vitro bioassays detected the estrogenic effects in environmental water samples (EEQ values in the range of 0.75-304 × EQS), although the concentrations of hormones were below the limit of quantification in analytical measurements. The present study confirms the applicability of reference materials for estrogenic effects' detection through bioassays and indicates possible methodological drawbacks of some of them that may lead to false negative/positive outcomes. The observed difference in responsiveness among bioassays - based on mixture composition - is probably due to biological differences between them, suggesting that panels of bioassays with different characteristics should be applied according to specific environmental pollution conditions.

Environmental Risk of Pesticides for Fish in Small- and Medium-Sized Streams of Switzerland.

This study assessed the acute and chronic risk of pesticides, singly and as mixtures, for fish using comprehensive chemical data of four monitoring studies conducted in small- and medium-sized streams of Switzerland between 2012 and 2018. Pesticides were ranked based on single substance risk quotients and relative contribution to mixture risk. Concentrations of the pyrethroid insecticides, λ-cyhalothrin, cypermethrin and deltamethrin, and the fungicides, carbendazim and fenpropimorph, posed acute or chronic single substance risks. Risk quotients of eighteen additional pesticides were equal or greater than 0.1, and thirteen of those contributed ≥30% to mixture risk. Relatively few substances dominated the mixture risk in most water samples, with chronic and acute maximum cumulative ratios never exceeding 5 and 7, respectively. A literature review of toxicity data showed that concentrations of several pesticides detected in Swiss streams were sufficient to cause direct sublethal effects on fish in laboratory studies. Based on the results of our study, we conclude that pesticides detected in Swiss streams, especially pyrethroid insecticides, fungicides and pesticide mixtures, pose a risk to fish health and can cause direct sublethal effects at environmental concentrations. Sensitive life stages of species with highly specialized life history traits may be particularly vulnerable; however, the lack of toxicity data for non-model species currently prevents a conclusive assessment across species.

Evaluation of an in vitro assay to screen for the immunotoxic potential of chemicals to fish.

A wide variety of environmental contaminants has been shown to disrupt immune functions of fish and may compromise their defense capability against pathogens. Immunotoxic effects, however, are rarely considered in ecotoxicological testing strategies. The aim of this study was to systematically evaluate the suitability of an in vitro immuno-assay using selected fish immune parameters to screen for chemicals with known immunotoxic potential and to differentiate them from non-immunotoxicants. Non-stimulated and lipopolysaccharide-stimulated head kidney leukocytes of rainbow trout (Oncorhynchus mykiss) were exposed for 3 h or 19 h to chemicals with different modes of action. As immune parameters, phagocytosis activity, oxidative burst activity and cytokine transcription (IL-1ß, TNFα, IL-10) were examined, accompanied by in silico modelling. The immunotoxicants dexamethasone, benzo(a)pyrene, ethinylestradiol and bisphenol A significantly altered the immune parameters at non-cytotoxic concentrations whereas diclofenac had only weak effects. However, the two baseline chemicals with no known immunotoxic potential, butanol and ethylene glycol, caused significant effects, too. From our results it appears that the in vitro fish leukocyte assay as performed in the present study has only a limited capacity for discriminating between immunotoxicants and non-immunotoxicants.

Recycled low-density polyethylene composite to mitigate the environmental impacts generated from coal mining waste in Brazil.

Waste materials from coal mining and consumer products can pose significant risks to the environment. Residual coal deposits lead to the formation of acid drainage and release of contaminants, causing negative changes in soil and aquatic systems. Low density polyethylene (LDPE) polymers are an environmental concern due to their high useage, and slow degradation in the environment. In this study both waste materials were used to develop a composite to mitigate the environmental impacts of coal mining waste (CMW). The composite material was produced in different formulations (0-80 % wt CMW), and samples were tested for formation of acid drainage and release of contaminants. Chemical characterisation of the CMW and leachate of the composite materials was performed by X-ray fluorescence and atomic absorption spectrometry. Ecotoxicological effects in soil and water were investigated using standard tests with the earthworm, Eisenia fetida, the collembolan, Folsomia candida and the bacterium, Aliivibrio fischeri. Composites with 20 % wt LDPE showed a 50% increase in the pH value of the leachate compared to the CMW leachate. Iron, aluminium and sulfate concentrations were lower in leachates of the composite materials, and a reduction in the ecotoxicological impact on the tested organisms was observed. The hydrophobic nature of the composite's polymeric matrix as well its physical properties contributed to a better coating of the coal residue particles, blocking the contact with water and reducing the environmental risks of CMW. These results show that the production of composite material is a viable alternative route for treating coal and LDPE waste.

Leg Dominance Effects on Postural Control When Performing Challenging Balance Exercises.

Leg dominance reflects the preferential use of one leg over another and is typically attributed to asymmetries in the neural circuitry. Detecting leg dominance effects on motor behavior, particularly during balancing exercises, has proven difficult. The current study applied a principal component analysis (PCA) on kinematic data, to assess bilateral asymmetry on the coordinative structure (hypothesis H1) or on the control characteristics of specific movement components (hypothesis H2). Marker-based motion tracking was performed on 26 healthy adults (aged 25.3 ± 4.1 years), who stood unipedally on a multiaxial unstable board, in a randomized order, on their dominant and non-dominant leg. Leg dominance was defined as the kicking leg. PCA was performed to determine patterns of correlated segment movements ("principal movements" PMks). The control of each PMk was characterized by assessing its acceleration (second-time derivative). Results were inconclusive regarding a leg-dominance effect on the coordinative structure of balancing movements (H1 inconclusive); however, different control (p = 0.005) was observed in PM3, representing a diagonal plane movement component (H2 was supported). These findings supported that leg dominance effects should be considered when assessing or training lower-limb neuromuscular control and suggest that specific attention should be given to diagonal plane movements.

High-throughput DNA barcoding of oligochaetes for abundance-based indices to assess the biological quality of sediments in streams and lakes.

Aquatic oligochaete communities are valuable indicators of the biological quality of sediments in streams and lakes, but identification of specimens to the species level based on morphological features requires solid expertise in taxonomy and is possible only for a fraction of specimens present in a sample. The identification of aquatic oligochaetes using DNA barcodes would facilitate their use in biomonitoring and allow a wider use of this taxonomic group for ecological diagnoses. Previous approaches based on DNA metabarcoding of samples composed of total sediments or pools of specimens have been proposed for assessing the biological quality of ecosystems, but such methods do not provide precise information on species abundance, which limits the value of resulting ecological diagnoses. Here, we tested how a DNA barcoding approach based on high-throughput sequencing of sorted and genetically tagged specimens performed to assess oligochaete species diversity and abundance and the biological quality of sediments in streams and lakes. We applied both molecular and morphological approaches at 13 sites in Swiss streams and at 7 sites in Lake Geneva. We genetically identified 33 or 66 specimens per site. For both approaches, we used the same index calculations. We found that the ecological diagnoses derived from the genetic approach matched well with those of the morphological approach and that the genetic identification of only 33 specimens per site provided enough ecological information for correctly estimating the biological quality of sediments in streams and lakes.

Complete transcriptome assembly and annotation of a critically important amphipod species in freshwater ecotoxicological risk assessment: Gammarus fossarum.

Because of their crucial role in ecotoxicological risk assessment, amphipods (Crustacea) are commonly employed as model species in a wide range of studies. However, despite their ecological importance, their genome has not yet been completely annotated and molecular mechanisms underlying key pathways, such as the serotonin pathway, in development of ecotoxicological biomarkers of exposure to neuroactive pharmaceuticals are still poorly understood. Furthermore, genetic similarities and discrepancies with other model arthropods (e.g., Drosophila melanogaster) have not been completely clarified. In this report, we present a new transcriptome assembly of Gammarus fossarum, an important amphipod species, widespread in Central Europe. RNA-Seq with Illumina HiSeq technology was used to analyse samples extracted from total internal tissues. We used the Trinity and Trinotate software suites for transcriptome assembly and annotation, respectively. The quality of this assembly and the affiliated targeted homology searches greatly enrich the molecular knowledge on this species. Because of the lack of publicly available molecular information on the serotonin pathway, we also highlighted sequence homologies and divergences of the genes encoding the serotonin pathway components of the well-annotated arthropod D. melanogaster, and Crustacea with the corresponding genes of our assembly. An inferior number of hits was found when running a BLAST analysis of both D. melanogaster and Crustacea mRNA sequences encoding serotonin receptors available in GenBank against the total assembly, compared to other serotonin pathway components. A lack of information on important components for serotonin biosynthesis and vesicle endocytosis (i.e., tryptophan hydroxylase and vesicular monoamine transporter) in Crustacea was also brought to light. Our results will provide an extensive transcriptional resource for this important species in ecotoxicological risk assessment and highlight the need for a more detailed categorization of neuronal pathways components in invertebrates.

Infant Leg Activity Intensity Before and After Naps.

Wearable sensors are being used to measure intensity of infant physical activity across full days. The variability of infant activity intensity within and across days is important to study given the potential impact of physical activity on developmental trajectories. Using retrospective data, we analyzed the intensity of leg movements in 10 typically-developing infants before and after naptimes. Leg movement data were captured from 20 minutes before and after multiple events of naps across seven days for each infant. We hypothesized that leg movement intensity would be lower before a nap than after a nap potentially due to lower arousal and increased fatigue prior to attaining sleep. However, our results showed that leg movement intensity was not significantly different when comparing the 20-minute period pre- and post-naps (F(1,7)=3.91, p=0.089, ηp 2=0.358). Our results are a first step in describing patterns of infant activity across days and highlights the need for further research regarding infant energy expenditure and physical activity.

How Do Movement Patterns in Weightlifting (Clean) Change When Using Lighter or Heavier Barbell Loads?-A Comparison of Two Principal Component Analysis-Based Approaches to Studying Technique.

This study compared whole body kinematics of the clean movement when lifting three different loads, implementing two data analysis approaches based on principal component analysis (PCA). Nine weightlifters were equipped with 39 markers and their motion captured with 8 Vicon cameras at 100 Hz. Lifts of 60, 85, and 95% of the one repetition maximum were analyzed. The first PCA (PCAtrial) analyzed variance among time-normed waveforms compiled from subjects and trials; the second PCA (PCAposture) analyzed postural positions compiled over time, subjects and trials. Load effects were identified through repeated measures ANOVAs with Bonferroni-corrected post-hocs and through Cousineau-Morey confidence intervals. PCAtrial scores differed in the first (p < 0.016, η p 2 = 0.694) and fifth (p < 0.006, η p 2 = 0.768) principal component, suggesting that increased barbell load produced higher initial elevation, lower squat position, wider feet position after squatting, and less inclined arms. PCAposture revealed significant timing differences in all components. We conclude, first, barbell load affects specific aspects of the movement pattern of the clean; second, the PCAtrial approach is better suited for detecting deviations from a mean motion trajectory and its results are easier to interpret; the PCAposture approach reveals coordination patterns and facilitates comparisons of postural speeds and accelerations.

Effects of treated wastewater on the ecotoxicity of small streams - Unravelling the contribution of chemicals causing effects.

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: 17ß-estradiol equivalent concentrations (EEQbio, 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 (DEQbio) 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 (PtEQbio) concentrations (indicating the degree of AChE inhibition) reached relatively high values (37 to 1278 ng/L) mostly exceeding the corresponding EBT (196 ng/L PtEQbio). 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.