Metal Exposure and Sex Shape the Fatty Acid Profile of Midges and Reduce the Aquatic Subsidy to Terrestrial Food Webs.

Aquatic micropollutants can be transported to terrestrial systems and their consumers by emergent aquatic insects. However, micropollutants, such as metals, may also affect the flux of physiologically important polyunsaturated fatty acids (PUFAs). As certain PUFAs have been linked to physiological fitness and breeding success of terrestrial consumers, reduced fluxes from aquatic systems could affect terrestrial populations and food webs. We chronically exposed larvae of the aquatic insect Chironomus riparius to a range of environmentally relevant sediment contents of cadmium (Cd) or copper (Cu) in a 28-day microcosm study. Since elevated water temperatures can enhance metals' toxic effects, we used two temperature regimes, control and periodically elevated temperatures (heat waves) reflecting an aspect of climate change. Cd and Cu significantly reduced adult emergence by up to 95% and 45%, respectively, while elevated temperatures had negligible effects. Both metal contents were strongly reduced (∼90%) during metamorphosis. Furthermore, the chironomid FA profile was significantly altered during metamorphosis with the factors sex and metal exposure being relevant predictors. Consequently, fluxes of physiologically important PUFAs by emergent adults were reduced by up to ∼80%. Our results suggest that considering fluxes of physiologically important compounds, such as PUFAs, by emergent aquatic insects is important to understand the implications of aquatic micropollutants on aquatic-terrestrial meta-ecosystems.

Physiological variables in machine learning QSARs allow for both cross-chemical and cross-species predictions.

A major challenge in ecological risk assessment is estimating chemical-induced effects across taxa without species-specific testing. Where ecotoxicological data may be more challenging to gather, information on species physiology is more available for a broad range of taxa. Physiology is known to drive species sensitivity but understanding about the relative contribution of specific underlying processes is still elusive. Consequently, there remains a need to understand which physiological processes lead to differences in species sensitivity. The objective of our study was to utilize existing knowledge about organismal physiology to both understand and predict differences in species sensitivity. Machine learning models were trained to predict chemical- and species-specific endpoints as a function of both chemical fingerprints/descriptors and physiological properties represented by dynamic energy budget (DEB) parameters. We found that random forest models were able to predict chemical- and species-specific endpoints, and that DEB parameters were relatively important in the models, particularly for invertebrates. Our approach illuminates how physiological properties may drive species sensitivity, which will allow more realistic predictions of effects across species without the need for additional animal testing.

Nitrogen retention in stream biofilms - A potential contribution to the self-cleaning capacity.

Surface waters are under increasing pressure due to human activities, such as nutrient emissions from wastewater treatment plants (WWTPs). Using the retention of nitrogen (N) released from WWTPs as a proxy, we assessed the contribution of biofilms grown on inorganic and organic substrates to the self-cleaning capacity of second-order streams within the biosphere reserve Vosges du Nord/Palatinate forest (France/Germany). The uptake of N from anthropogenic sources, which is enriched with the heavy isotope 15N, into biofilms was assessed up- and downstream of WWTPs after five weeks of substrate deployment. Biofilms at downstream sites showed a significant positive linear relationship between δ15N and the relative contribution of wastewater to the streams' discharge. Furthermore, δ15N substantially increased in areas affected by WWTP effluent (∼8.5‰ and ∼7‰ for inorganic and organic substrate-associated biofilms, respectively) and afterwards declined with increasing distance to the WWTP effluent, approaching levels of upstream sections. The present study highlights that biofilms contribute to nutrient retention and likely the self-cleaning capacity of streams. This function seems, however, to be limited by the fact that biofilms are restricted in their capacity to process excessive N loads with large differences between individual reaches (e.g., δ15N: -3.25 to 12.81‰), influenced by surrounding conditions (e.g., land use) and modulated through climatic factors and thus impacted by climate change. Consequently, the impact of WWTPs located close to the source of a stream are dampened by the biofilms' capacity to retain N only to a minor share and suggest substantial N loads being transported downstream.

Trophic Dynamics of Mercury in the Baltic Archipelago Sea Food Web: The Impact of Ecological and Ecophysiological Traits.

We investigated trophic dynamics of Hg in the polluted Baltic Archipelago Sea using established trophic magnification (TMFs) and biomagnification factors (BMFs) on a comprehensive set of bird, fish, and invertebrate species. As different ecological and ecophysiological species traits may affect trophic dynamics, we explored the effect of food chain (benthic, pelagic, benthopelagic) and thermoregulatory strategy on trophic total Hg (THg) dynamics, using different approaches to accommodate benthopelagic species and normalize for trophic position (TP). We observed TMFs and most BMFs greater than 1, indicating overall THg biomagnification. We found significantly higher pelagic TMFs (3.58-4.02) compared to benthic ones (2.11-2.34) when the homeotherm bird species were excluded from models, but not when included. This difference between the benthic and pelagic TMFs remained regardless of how the TP of benthopelagic species was modeled, or whether TMFs were normalized for TP or not. TP-corrected BMFs showed a larger range (0.44-508) compared to BMFs representing predator-prey concentration ratios (0.05-82.2). Overall, the present study shows the importance of including and evaluating the effect of ecological and ecophysiological traits when investigating trophic contaminant dynamics.

Legacy and emerging organohalogenated compounds in feathers of Eurasian eagle-owls (Bubo bubo) in Norway: Spatiotemporal variations and associations with dietary proxies (δ13C and δ15N).

The occurrence of organohalogenated compounds (OHCs) in wildlife has received considerable attention over the last decades. Among the matrices used for OHCs biomonitoring, feathers are particularly useful as they can be collected in a minimally or non-invasive manner. In this study, concentrations of various legacy OHCs -polychlorinated biphenyls (PCBs), organochlorine pesticides (OCPs) and polybrominated diphenyl ethers (PBDEs)-, as well as emerging OHCs -per- and polyfluoroalkyl substances (PFAS) and organophosphate ester flame retardants (OPEs)- were determined in feathers of 72 Eurasian eagle-owls (Bubo bubo) from Norway, with the goal of studying spatiotemporal variation using a non-invasive approach. Molted feathers were collected at nest sites from northern, central and southern Norway across four summers (2013-2016). Additionally, two museum-archived feathers from 1979 to 1989 were included. Stable carbon (δ13C) and nitrogen isotopes (δ15N) were used as dietary proxies. In total, 11 PFAS (sum range 8.25-215.90 ng g-1), 15 PCBs (4.19-430.01 ng g-1), 6 OCPs (1.48-220.94 ng g-1), 5 PBDEs (0.21-5.32 ng g-1) and 3 OPEs (4.49-222.21 ng g-1) were quantified. While we observed large variation in the values of both stable isotopes, suggesting a diverse diet of the eagle-owls, only δ13C seemed to explain variation in PFAS concentrations. Geographic area and year were influential factors for δ15N and δ13C. Considerable spatial variation was observed in PFAS levels, with the southern area showing higher levels compared to northern and central Norway. For the rest of OHCs, we observed between-year variations; sum concentrations of PCBs, OCPs, PBDEs and OPEs reached a maximum in 2015 and 2016. Concentrations from 1979 to 1989 were within the ranges observed between 2013 and 2016. Overall, our data indicate high levels of legacy and emerging OHCs in a top predator in Norway, further highlighting the risk posed by OHCs to wildlife.

Herbicide-Induced Shifts in the Periphyton Community Composition Indirectly Affect Feeding Activity and Physiology of the Gastropod Grazer Physella acuta.

Herbicides are well known for unintended effects on freshwater periphyton communities. Large knowledge gaps, however, exist regarding indirect herbicide impacts on primary consumers through changes in the quality of periphyton as a food source (i.e., diet-related effects). To address this gap, the grazer Physella acuta (Gastropoda) was fed for 21 days with periphyton that grew for 15 days in the presence or absence of the herbicide diuron (8 µg/L) to quantify changes in the feeding rate, growth rate, and energy storage (neutral lipid fatty acids; NLFAs) of P. acuta. Periphyton biomass, cell viability, community structure, and FAs served as proxies for food quality that support a mechanistic interpretation of the grazers' responses. Diuron changed the algae periphyton community and fatty acid profiles, indicating alterations in the food quality, which could explain differences in the snails' feeding rate compared to the control. While the snails' growth rate was, despite an effect size of 55%, not statistically significantly changed, NLFA profiles of P. acuta were altered. These results indicate that herbicides can change the food quality of periphyton by shifts in the algae composition, which may affect the physiology of grazers.

Multiple Stressors in Aquatic Ecosystems: Sublethal Effects of Temperature, Dissolved Organic Matter, Light and a Neonicotinoid Insecticide on Gammarids.

Whether and to which extent the effects of chemicals in the environment interact with other factors remains a scientific challenge. Here we assess the combined effects of temperature (16 vs. 20°C), light conditions (darkness vs. 400 lx), dissolved organic matter (DOM; 0 vs. 6 mg/L) and the model insecticide thiacloprid (0 vs. 3 µg/L) in a full-factorial experiment on molting and leaf consumption of Gammarus fossarum. Thiacloprid was the only factor significantly affecting gammarids' molting. While DOM had low effects on leaf consumption, temperature, light and thiacloprid significantly affected this response variable. The various interactions among these factors were not significant suggesting additivity. Only the interaction of the factors temperature and thiacloprid suggested a tendency for antagonism. As most stressors interacted additively, their joint effects may be predictable with available models. However, synergistic interactions are difficult to capture while being central for securing ecosystem integrity.

The Fungicide Tebuconazole Confounds Concentrations of Molecular Biomarkers Estimating Fungal Biomass.

Due to their ecological importance, fungi are suitable indicator organisms for anthropogenic stress. To estimate fungal biomass, the fungal membrane molecule ergosterol is often quantified as a proxy. Estimates based on ergosterol may, however, be distorted by exposure to demethylase inhibiting (DMI) fungicides, interfering with sterol synthesis. To test this hypothesis, we exposed ten fungal species to the DMI fungicide tebuconazole and measured concentrations of ergosterol and DNA per unit dry mass of the fungal hyphae. The latter served as alternative biomass proxy that is not specifically targeted by tebuconazole. Effects of tebuconazole on ergosterol concentrations were species-specific, while concentrations were on average reduced by 13%. In contrast, DNA concentrations were on average increased by 13%. We demonstrate that DMI fungicides - at close to field relevant levels - can distort fungal biomass estimation, complicating the use of this endpoint for environmental management.

Fungicides: An Overlooked Pesticide Class?

Fungicides are indispensable to global food security and their use is forecasted to intensify. Fungicides can reach aquatic ecosystems and occur in surface water bodies in agricultural catchments throughout the entire growing season due to their frequent, prophylactic application. However, in comparison to herbicides and insecticides, the exposure to and effects of fungicides have received less attention. We provide an overview of the risk of fungicides to aquatic ecosystems covering fungicide exposure (i.e., environmental fate, exposure modeling, and mitigation measures) as well as direct and indirect effects of fungicides on microorganisms, macrophytes, invertebrates, and vertebrates. We show that fungicides occur widely in aquatic systems, that the accuracy of predicted environmental concentrations is debatable, and that fungicide exposure can be effectively mitigated. We additionally demonstrate that fungicides can be highly toxic to a broad range of organisms and can pose a risk to aquatic biota. Finally, we outline central research gaps that currently challenge our ability to predict fungicide exposure and effects, promising research avenues, and shortcomings of the current environmental risk assessment for fungicides.

White-Tailed Eagle (Haliaeetus albicilla) Body Feathers Document Spatiotemporal Trends of Perfluoroalkyl Substances in the Northern Environment.

We reconstructed the first long-term (1968-2015) spatiotemporal trends of perfluoroalkyl substances (PFAS) using archived body feathers of white-tailed eagles (Haliaeetus albicilla) from the West Greenland (n = 31), Norwegian (n = 66), and Central Swedish Baltic coasts (n = 50). We observed significant temporal trends of perfluorooctane sulfonamide (FOSA), perfluorooctane sulfonate (PFOS), and perfluoroalkyl carboxylates (∑PFCAs) in all three subpopulations. Concentrations of FOSA and PFOS had started decreasing significantly since the mid-1990s to 2000 in the Greenland and Norwegian subpopulations, consistent with the 3M phase-out, though in sharp contrast to overall increasing trends observed in the Swedish subpopulation. Moreover, ∑PFCA concentrations significantly increased in all three subpopulations throughout the study periods. These temporal trends suggest on-going input of PFOS in the Baltic and of ∑PFCAs in all three regions. Considerable spatial variation in PFAS concentrations and profiles was observed: PFOS concentrations were significantly higher in Sweden, whereas FOSA and ∑PFCA concentrations were similar among the subpopulations. PFOS dominated the PFAS profiles in the Swedish and Norwegian subpopulations, in contrast to the domination of FOSA and ∑PFCAs in the Greenland one. Our spatiotemporal observations underline the usefulness of archived bird of prey feathers in monitoring spatiotemporal PFAS trends and urge for continued monitoring efforts in each of the studied subpopulations.

Is Hyalella azteca a Suitable Model Leaf-Shredding Benthic Crustacean for Testing the Toxicity of Sediment-Associated Metals in Europe?

The leaf-shredding crustacean Hyalella azteca, which is indigenous to Northern and Central America, is used to assess environmental risks associated with (metal-)contaminated sediments and to propose sediment quality standards also in Europe. Yet, it is unknown if H. azteca is protective for European crustacean shredders. We thus compared the sensitivity of H. azteca with that of the European species Asellus aquaticus and Gammarus fossarum towards copper- and cadmium-contaminated sediments (prepared according to OECD 218) under laboratory conditions employing mortality and leaf consumption as endpoints. H. azteca either reacted approximately fourfold more sensitive than the most tolerant tested species (as for cadmium) or its sensitivity was only 1.6 times lower than the highest sensitivity determined (as for copper), which should be covered by safety factors applied during risk assessments. Therefore, the results for the sediment type and the two heavy metals tested during the present study in combination with the existence of standardized testing protocols, their ease of culture, and short generation time, suggest H. azteca as suitable crustacean model shredder for assessing the toxicity of sediment-associated metals in Europe.

History matters: Heterotrophic microbial community structure and function adapt to multiple stressors.

Ecosystem functions in streams (e.g., microbially mediated leaf litter breakdown) are threatened globally by the predicted agricultural intensification and its expansion into pristine areas, which is associated with increasing use of fertilizers and pesticides. However, the ecological consequences may depend on the disturbance history of microbial communities. To test this, we assessed the effects of fungicides and nutrients (four levels each) on the structural and functional resilience of leaf-associated microbial communities with differing disturbance histories (pristine vs. previously disturbed) in a 2 × 4 × 4-factorial design (n = 6) over 21 days. Microbial leaf breakdown was assessed as a functional variable, whereas structural changes were characterized by the fungal community composition, species richness, biomass, and other factors. Leaf breakdown by the pristine microbial community was reduced by up to 30% upon fungicide exposure compared with controls, whereas the previously disturbed microbial community increased leaf breakdown by up to 85%. This significant difference in the functional response increased in magnitude with increasing nutrient concentrations. A pollution-induced community tolerance in the previously disturbed microbial community, which was dominated by a few species with high breakdown efficacies, may explain the maintained function under stress. Hence, the global pressure on pristine ecosystems by agricultural expansion is expected to cause a modification in the structure and function of heterotrophic microbial communities, with microbially mediated leaf litter breakdown likely becoming more stable over time as a consequence of fungal community adaptions.

Modeling Remobilization of Neonicotinoid Residues from Tree Foliage in Streams-A Relevant Exposure Pathway in Risk Assessment?

Systemic neonicotinoid insecticides are increasingly used as a crop protection measure to suppress insect pests on trees. However, senescent foliage falling from treated trees represents a rarely studied pathway through which neonicotinoids may enter nontarget environments, e.g., surface waters. To estimate risk posed by this pathway, neonicotinoid residues were analyzed in foliage from black alder trees treated with one of three neonicotinoid insecticides (imidacloprid, thiacloprid, or acetamiprid) at five concentrations, each ranging from 0.0375-9.6 g active ingredient/cm trunk diameter at breast height (n = 3). Foliar residues measured at the time of leaf fall were used as input parameters for a model predicting imidacloprid water concentrations over a 100-m-long stream stretch as a consequence of remobilization from introduced foliage (input: 600 g foliage/m2 containing 80 µg imidacloprid/g). The water concentration (up to ∼250 ng/L) predicted by the model exceeded the recently proposed Maximum Permissible Concentration of 8.3 ng/L for ∼6.5 days. Moreover, dietary uptake was identified as an additional exposure route for aquatic organisms. The alternative pathway (i.e., introduction via leaf fall) and exposure route (i.e., dietary uptake) associated with the systemic nature of neonicotinoids should be accounted for during their registration process in order to safeguard ecosystem integrity.

Does Waterborne Exposure Explain Effects Caused by Neonicotinoid-Contaminated Plant Material in Aquatic Systems?

Neonicotinoids are increasingly applied on trees as protection measure against insect pests. Consequently, neonicotinoids are inevitably transferred into aquatic environments either via spray drift or surface runoff or (due to neonicotinoids' systemic nature) via senescent leaves. There particularly leaf-shredding invertebrates may be exposed to neonicotinoids through both the water phase and the consumption of contaminated leaves. In 7 day bioassays (n = 30), we examined ecotoxicological differences between these two exposure scenarios for an amphipod and an insect nymph with their feeding rate as the response variable. Organisms either experienced waterborne neonicotinoid (i.e., imidacloprid, thiacloprid, and acetamiprid) exposure only or a combined exposure (waterborne and dietary) through both the consumption of contaminated leaves and neonicotinoids leaching from leaves into water. The amphipod (7 day EC50s from 0.3 to 8.4 µg/L) was more sensitive than the insect nymph (7 day EC50s from 7.0 to 19.4 µg/L). Moreover, for both species, concentration-response models derived from water concentrations indicated higher effects under the combined exposure. Together with the observed inability of shredders to avoid neonicotinoid-contaminated leaves, our results emphasize the relevance of dietary exposure (e.g., via leaves) for systemic insecticides. Thus, it would be prudent to consider dietary exposure during the registration of systemic insecticides to safeguard ecosystem integrity.

History Matters: Pre-Exposure to Wastewater Enhances Pesticide Toxicity in Invertebrates.

Disturbance regimes determine communities' structure and functioning. Nonetheless, little effort has been undertaken to understand interactions of press and pulse disturbances. In this context, leaf-shredding macroinvertebrates can be chronically exposed to wastewater treatment plant effluents (i.e., press disturbance) before experiencing pesticide exposure following agricultural runoff (i.e., pulse disturbance). It is assumed that wastewater pre-exposure alters animals' sensitivity to pesticides. To test this hypothesis, we exposed model-populations of the shredder Gammarus fossarum to wastewater at three field-relevant dilution levels (i.e., 0%, 50%, and 100%). After 2, 4, and 6 weeks, survival, leaf consumption, dry weight, and energy reserves were monitored. Additionally, animals were assessed for their sensitivity toward the neonicotinoid insecticide thiacloprid using their feeding rate as response variable. Both wastewater treatments reduced gammarids' survival, leaf consumption, dry weight, and energy reserves. Moreover, both wastewater pre-exposure scenarios increased animals' sensitivity toward thiacloprid by up to 2.5 times compared to the control. Our results thus demonstrate that press disturbance as posed by wastewater pre-exposure can enhance susceptibility of key players in ecosystem functioning to further (pulse) disturbances. Therefore, applying mitigation measures such as advanced treatment technologies seems sensible to support functional integrity in the multiple-stress situation.

Does the current fungicide risk assessment provide sufficient protection for key drivers in aquatic ecosystem functioning?

The level of protection provided by the present environmental risk assessment (ERA) of fungicides in the European Union for fungi is unknown. Therefore, we assessed the structural and functional implications of five fungicides with different modes of action (azoxystrobin, carbendazim, cyprodinil, quinoxyfen, and tebuconazole) individually and in mixture on communities of aquatic hyphomycetes. This is a polyphyletic group of fungi containing key drivers in the breakdown of leaf litter, governing both microbial leaf decomposition and the palatability of leaves for leaf-shredding macroinvertebrates. All fungicides impaired leaf palatability to the leaf-shredder Gammarus fossarum and caused structural changes in fungal communities. In addition, all compounds except for quinoxyfen altered microbial leaf decomposition. Our results suggest that the European Union's first-tier ERA provides sufficient protection for the tested fungicides, with the exception of tebuconazole and the mixture, while higher-tier ERA does not provide an adequate level of protection for fungicides in general. Therefore, our results show the need to incorporate aquatic fungi as well as their functions into ERA testing schemes to safeguard the integrity of aquatic ecosystems.

Bio-QSARs 2.0: Unlocking a new level of predictive power for machine learning-based ecotoxicity predictions by exploiting chemical and biological information.

Practical, legal, and ethical reasons necessitate the development of methods to replace animal experiments. Computational techniques to acquire information that traditionally relied on animal testing are considered a crucial pillar among these so-called new approach methodologies. In this light, we recently introduced the Bio-QSAR concept for multispecies aquatic toxicity regression tasks. These machine learning models, trained on both chemical and biological information, are capable of both cross-chemical and cross-species predictions. Here, we significantly extend these models' applicability. This was realized by increasing the quantity of training data by a factor of approximately 20, accomplished by considering both additional chemicals and aquatic organisms. Additionally, variable test durations and associated random effects were accommodated by employing a machine learning algorithm that combines tree-boosting with mixed-effects modeling (i.e., Gaussian Process Boosting). We also explored various biological descriptors including Dynamic Energy Budget model parameters, taxonomic distances, as well as genus-specific traits and investigated the inclusion of mode-of-action information. Through these efforts, we developed Bio-QSARs for fish and aquatic invertebrates with exceptional predictive power (R squared of up to 0.92 on independent test sets). Moreover, we made considerable strides to make models applicable for a range of use cases in environmental risk assessment as well as research and development of chemicals. Models were made fully explainable by implementing an algorithmic multicollinearity correction combined with SHapley Additive exPlanations. Furthermore, we devised novel approaches for applicability domain construction that take feature importance into account. We are hence confident these models, which are available via open access, will make a significant contribution towards the implementation of new approach methodologies and ultimately have the potential to support "Green Chemistry" and "Green Toxicology".

Elevated Fungicide and Nutrient Concentrations Change Structure but not Function of Aquatic Microbial Communities.

Leaf decomposition is a key process in stream ecosystems within forested catchments; it is driven by microbial communities, particularly fungi and bacteria. These microorganisms make nutrients and energy bound in leaves available for wider parts of the food web. Leaf-associated microorganisms are subjected to anthropogenic pressures, such as the increased exposure to nutrients and fungicides associated with land-use change. We assessed the sensitivity of leaf-associated microbial communities with differing exposure histories, namely, from pristine (P) streams, and streams impacted by wastewater (W) and agricultural run-off (vineyards; V). In the laboratory, microbial communities were exposed to elevated nutrient (NO3-N: 0.2-18.0 mg/L, PO4-P: 0.02-1.8 mg/L) and fungicide concentrations (sum concentration 0-300 µg/L) in a fully crossed 3 × 4 × 4-factorial design over 21 days. Leaf decomposition and exoenzyme activity were measured as functional endpoints, and fungal community composition and microbial abundance served as structural variables. Overall, leaf decomposition did not differ between fungicide treatments or exposure histories. Nonetheless, substantial changes in the fungal community composition were observed after exposure to environmentally relevant fungicide concentrations. Elevated nutrient concentrations assisted leaf decomposition, and the effect size depended on the exposure history. The observed changes in the fungal community composition support the principle of functional redundancy, with highly efficient decomposers maintaining leaf decomposition. Environ Toxicol Chem 2024;43:1300-1311. © 2024 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Effects of peak exposure scenarios on Gammarus fossarum using field relevant pesticide mixtures.

The present study investigated sublethal effects of a field relevant pesticide mixture (one herbicide, three fungicides, five insecticides) on Gammarus fossarum by considering different peak exposure scenarios, which may be generated by the inherent properties of vegetated ditches. Additional experiments aimed at the identification of germane exposure pathways (food and water). Therefore, G. fossarum were exposed in independent experiments to three scenarios, which differed besides in the peak concentration of the pesticide mixture also in the mixture's composition and exposure duration (n=20 per treatment). The exposure duration of 12 or 120 min was followed by a seven-day post-exposure observation period. At a constant concentration-time product, a lower exposure duration in concert with a proportionally higher peak concentration caused a substantially elevated ecotoxicity compared to a treatment with a longer exposure duration at a lower peak concentration. Given the importance of the insecticide lambda-cyhalothrin for the mixture's ecotoxicity it may be concluded that the fast mode of action of pyrethroids mainly explains this observation. Moreover, field relevant concentrations of the pesticide mixture applied at an exposure duration of 120 min resulted in reduced gammarids' feeding rate, which may be indicative for shifts in the ecosystem function of leaf litter breakdown and hence the provision of energy for local and downstream communities. Finally, the present study indicated that both pathways of exposure, namely via food or water, reduce gammarids' feeding rate synergistically. This suggests that both exposure pathways should be considered for compounds exhibiting a high Kow (e.g. pyrethroids) during the risk assessment of single substances and mixtures.

Tracer or toxicant: Does stable isotope labeling affect central processes in aquatic food webs?

Stable isotope analysis (SIA) is an elementary technique in food web ecology, but its insights become increasingly ambiguous in complex systems. One approach to elevate the utility of SIA in such systems is the use of heavy isotope tracers (i.e., labeling). However, the fundamental assumption that the addition of such tracers does not affect in situ conditions has been challenged. This study tests if labeling is suitable for autotrophy-based and detritus-based aquatic food webs. For the former, the survival and reproduction of Daphnia magna fed with phytoplankton cultured at different levels of 15N addition were assessed. For the latter, the microbial decomposition of leaf litter was assessed at the same tracer levels. While no significant differences were observed, effect patterns were comparable to a previous study, supporting the isotopic redundancy hypothesis that postulates discrete quantum mechanical states at which the reaction speeds of metabolic processes are altered. Although physiology (reproduction) and activity (microbial decomposition) might not be altered to an ecologically significant level, labeling with heavy stable isotopes could potentially affect isotopic fractionation in biochemical processes and bias conclusions drawn from resulting SI ratios.