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.

Aging of nanosized titanium dioxide modulates the effects of dietary copper exposure on Daphnia magna - an assessment over two generations.

Nanosized titanium dioxide (nTiO2) is widely used in products, warranting its discharge from various sources into surface water bodies. However, nTiO2 co-occurs in surface waters with other contaminants, such as metals. Studies with nTiO2 and metals have indicated that the presence of natural organic matter (NOM) can mitigate their toxicity to aquatic organisms. In addition, "aging" of nTiO2 can affect toxicity. However, it is a research challenge, particularly when addressing sublethal responses from dietary exposure over multiple generations. We, therefore exposed the alga Desmodesmus subspicatus to nTiO2 (at concentrations of 0.0, 0.6 and 3.0 mg nTiO2/L) in nutrient medium aged for 0 or 3 days with copper (Cu) at concentrations of 0 and 116 µg Cu/L and with NOM at concentrations equivalent to 0 and 8 mg total organic carbon (TOC) per litre. Subsequently, the exposed alga was fed to Daphnia magna for 23 days over two generations and survival, reproduction and body length were assessed as endpoints of toxicity. In parallel, Cu accumulation and depuration from D. magna were measured. The results indicate that the reproduction of D. magna was the most sensitive parameter in this study, being reduced by 30% (at both parental (F0) and filial (F1) generations) and 50% (at F0 but not F1) due to the dietary Cu exposure in combination with nTiO2 for 0 and 3 days aging, respectively. There was no relationship between the effects observed on reproduction and Cu body burden in D. magna. Moreover, D. magna from the F1 generation showed an adaptive response to Cu in the treatment with 3.0 mg nTiO2/L aged for 3 days, potentially due to epigenetic inheritance. Unexpectedly, the presence of NOM hardly changed the observed effects, pointing towards the function of algal exopolymeric substances or intracellular organic matter, rendering the NOM irrelevant. Ultimately, the results indicate that the transferability of the impacts observed during the F0 to the responses in the F1 generation is challenging due to opposite effect directions. Additional mechanistic studies are needed to unravel this inconsistency in the responses between generations and to support the development of reliable effect models.

Microbial community history and leaf species shape bottom-up effects in a freshwater shredding amphipod.

Arable land use and the associated application of agrochemicals can affect local freshwater communities with consequences for the entire ecosystem. For instance, the structure and function of leaf-associated microbial communities can be affected by pesticides, such as fungicides. Additionally, the leaf species on which these microbial communities grow reflects another environmental filter for community structure. These factors and their interaction may jointly modify leaves' nutritional quality for higher trophic levels. To test this assumption, we studied the structure of leaf-associated microbial communities with distinct exposure histories (pristine [P] vs vineyard run off [V]) colonising two leaf species (black alder, European beech, and a mixture thereof). By offering these differently colonised leaves as food to males and females of the leaf-shredding amphipod Gammarus fossarum (Crustacea; Amphipoda) we assessed for potential bottom-up effects. The growth rate, feeding rate, faeces production and neutral lipid fatty acid profile of the amphipod served as response variable in a 2 × 3 × 2-factorial test design over 21d. A clear separation of community history (P vs V), leaf species and an interaction between the two factors was observed for the leaf-associated aquatic hyphomycete (i.e., fungal) community. Sensitive fungal species were reduced by up to 70 % in the V- compared to P-community. Gammarus' growth rate, feeding rate and faeces production were affected by the factor leaf species. Growth was negatively affected when Gammarus were fed with beech leaves only, whereas the impact of alder and the mixture of both leaf species was sex-specific. Overall, this study highlights that leaf species identity had a more substantial impact on gammarids relative to the microbial community itself. Furthermore, the sex-specificity of the observed effects (excluding fatty acid profile, which was only measured for male) questions the procedure of earlier studies, that is using either only one sex or not being able to differentiate between males and females. However, these results need additional verification to support a reliable extrapolation.

Effects of flow reduction and artificial light at night (ALAN) on litter decomposition and invertebrate communities in streams: A flume experiment.

River ecosystems are heavily impacted by multiple stressors, where effects can cascade downstream of point sources. However, a spatial approach to assess the effects of multiple stressors is missing. We assessed the local and downstream effects on litter decomposition, and associated invertebrate communities of two stressors: flow reduction and artificial light at night (ALAN). We used an 18-flow-through mesocosm system consisting of two tributaries, where we applied the stressors, merging in a downstream section. We assessed the changes in decomposition rate and invertebrate community structure in leaf bags. We found no effect of ALAN or its interaction with flow reduction on the litter decomposition or the invertebrate community in the tributaries. Flow reduction alone led to a 14.8 % reduction in decomposition rate in the tributaries. We recorded no effect of flow reduction on the macroinvertebrates community composition in the litter bags. We also observed no effects of the spatial arrangement of the stressors on the litter decomposition and macroinvertebrate community structure downstream. Overall, our results suggest the impact of stressors on litter decomposition and macroinvertebrate communities remained local in our experiment. Our work thus calls for further studies to identify the mechanisms and the conditions under which spatial effects dominate over local processes.

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.

Diatoms Reduce Decomposition of and Fungal Abundance on Less Recalcitrant Leaf Litter via Negative Priming.

Heterotrophic microbial decomposers colonize submerged leaf litter in close spatial proximity to periphytic algae that exude labile organic carbon during photosynthesis. These exudates are conjectured to affect microbial decomposers' abundance, resulting in a stimulated (positive priming) or reduced (negative priming) leaf litter decomposition. Yet, the occurrence, direction, and intensity of priming associated with leaf material of differing recalcitrance remains poorly tested. To assess priming, we submerged leaf litter of differing recalcitrance (Alnus glutinosa [alder; less recalcitrant] and Fagus sylvatica [beech; more recalcitrant]) in microcosms and quantified bacterial, fungal, and diatom abundance as well as leaf litter decomposition over 30 days in absence and presence of light. Diatoms did not affect beech decomposition but reduced alder decomposition by 20% and alder-associated fungal abundance by 40% in the treatments including all microbial groups and light, thus showing negative priming. These results suggest that alder-associated heterotrophs acquired energy from diatom exudates rather than from leaf litter. Moreover, it is suggested that these heterotrophs have channeled energy to alternative (reproductive) pathways that may modify energy and nutrient availability for the remaining food web and result in carbon pools protected from decomposition in light-exposed stream sections.

Leaf Species-Dependent Fungicide Effects on the Function and Abundance of Associated Microbial Communities.

Microbially-mediated leaf litter decomposition is a critical ecosystem function in running waters within forested areas, which can be affected by fungicides. However, fungicide effects on leaf litter decomposition have been investigated almost exclusively with black alder leaves, a leaf species with traits favourable to consumers (i.e., low recalcitrance and high nutrient content). At the same time, little is known about fungicide effects on microbial colonisation and decomposition of other leaf species with less favourable traits. In this 21 day lasting study, we explore the effects of increasing fungicide sum concentrations (0-3000 µg/L) on microbial colonisation and decomposition of three leaf species (black alder, Norway maple and European beech) differing in terms of recalcitrance and nutrient content. Leaf litter decomposition rate, leaf-associated fungal biomass and bacterial density were quantified to observe potential effects at the functional level. Beech, as the species with the least favourable leaf traits, showed a substantially lower decomposition rate (50%) in absence of fungicides than alder and maple. In the presence of high fungicide concentrations (300-3000 µg/L), beech showed a concentration-related decrease not only in microbial leaf litter decomposition but also fungal biomass. This suggests that favourable traits of leaf litter (as for alder and maple) enable leaf-associated microorganisms to acquire leaf-bound energy more easily to withstand potential effects induced by fungicide exposure. Our results indicate the need to deepen our understanding on how leaf species' traits interact with the impact of chemical stressors on the leaf decomposition activity of microbial communities.

Interactions between titanium dioxide nanoparticles and polyethylene microplastics: Adsorption kinetics, photocatalytic properties, and ecotoxicity.

The present study investigated the adsorption mechanism of titanium dioxide nanoparticles (nTiO2) on polyethylene microplastics (MPs) and the resulting photocatalytic properties. This effort was supported by ecotoxicological assessments of MPs with adsorbed nTiO2 on the immobility and behaviour of Daphnia magna in presence and absence of UV irradiation. The results showed that nTiO2 were rapidly adsorbed on the surface of MPs (72% of nTiO2 in 9 h). The experimental data fit well with the pseudo-second order kinetic model. Both suspended nTiO2 and nTiO2 immobilized on MPs exhibited comparable photocatalytic properties, with the latter showing a lower effect on Daphnia mobility. A likely explanation is that the suspended nTiO2 acted as a homogeneous catalyst under UV irradiation and generated hydroxyl radicals throughout the test vessel, whereas the nTiO2 adsorbed on MPs acted as a heterogeneous catalyst and generated hydroxyl radicals only locally and thus near the air-water interface. Consequently, Daphnia, which were hiding at the bottom of the test vessel, actively avoided exposure to hydroxyl radicals. These results suggest that the presence of MPs can modulate the phototoxicity of nTiO2 - at least the location at which it is active - under the studied conditions.

Standard Versus Natural: Assessing the Impact of Environmental Variables on Organic Matter Decomposition in Streams Using Three Substrates.

The decomposition of allochthonous organic matter, such as leaves, is a crucial ecosystem process in low-order streams. Microbial communities, including fungi and bacteria, colonize allochthonous organic material, break up large molecules, and increase the nutritional value for macroinvertebrates. Environmental variables are known to affect microbial as well as macroinvertebrate communities and alter their ability to decompose organic matter. Studying the relationship between environmental variables and decomposition has mainly been realized using leaves, with the drawbacks of differing substrate composition and consequently between-study variability. To overcome these drawbacks, artificial substrates have been developed, serving as standardizable surrogates. In the present study, we compared microbial and total decomposition of leaves with the standardized substrates of decotabs and, only for microbial decomposition, of cotton strips, across 70 stream sites in a Germany-wide study. Furthermore, we identified the most influential environmental variables for the decomposition of each substrate from a range of 26 variables, including pesticide toxicity, concentrations of nutrients, and trace elements, using stability selection. The microbial as well as total decomposition of the standardized substrates (i.e., cotton strips and decotabs) were weak or not associated with that of the natural substrate (i.e., leaves, r² < 0.01 to r² = 0.04). The decomposition of the two standardized substrates, however, showed a moderate association (r² = 0.21), which is probably driven by their similar composition, with both being made of cellulose. Different environmental variables were identified as the most influential for each of the substrates and the directions of these relationships contrasted between the substrates. Our results imply that these standardized substrates are unsuitable surrogates when investigating the decomposition of allochthonous organic matter in streams. Environ Toxicol Chem 2023;42:2007-2018. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Structural and functional development of twelve newly established floodplain pond mesocosms.

Ecosystems are complex structures with interacting abiotic and biotic processes evolving with ongoing succession. However, limited knowledge exists on the very initial phase of ecosystem development and colonization. Here, we report results of a comprehensive ecosystem development monitoring for twelve floodplain pond mesocosms (FPM; 23.5 m × 7.5 m × 1.5 m each) located in south-western Germany. In total, 20 abiotic and biotic parameters, including structural and functional variables, were monitored for 21 months after establishment of the FPMs. The results showed evolving ecosystem development and primary succession in all FPMs, with fluctuating abiotic conditions over time. Principal component analyses and redundancy analyses revealed season and succession time (i.e., time since ecosystem establishment) to be significant drivers of changes in environmental conditions. Initial colonization of both aquatic (i.e., water bodies) and terrestrial (i.e., riparian land areas) parts of the pond ecosystems occurred within the first month, with subsequent season-specific increases in richness and abundance for aquatic and terrestrial taxa over the entire study period. Abiotic environmental conditions and aquatic and terrestrial communities showed increasing interpond variations over time, that is, increasing heterogeneity among the FPMs due to natural environmental divergence. However, both functional variables assessed (i.e., aquatic and terrestrial litter decomposition) showed opposite patterns as litter decomposition rates slightly decreased over time and interpond differences converged with successional ecosystem developments. Overall, our results provide rare insights into the abiotic and biotic conditions and processes during the initial stages of freshwater ecosystem formation, as well as into structural and functional developments of the aquatic and terrestrial environment of newly established pond ecosystems.

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.

Forest streams are important sources for nitrous oxide emissions.

Streams and river networks are increasingly recognized as significant sources for the greenhouse gas nitrous oxide (N2 O). N2 O is a transformation product of nitrogenous compounds in soil, sediment and water. Agricultural areas are considered a particular hotspot for emissions because of the large input of nitrogen (N) fertilizers applied on arable land. However, there is little information on N2 O emissions from forest streams although they constitute a major part of the total stream network globally. Here, we compiled N2 O concentration data from low-order streams (~1,000 observations from 172 stream sites) covering a large geographical gradient in Sweden from the temperate to the boreal zone and representing catchments with various degrees of agriculture and forest coverage. Our results showed that agricultural and forest streams had comparable N2 O concentrations of 1.6 ± 2.1 and 1.3 ± 1.8 µg N/L, respectively (mean ± SD) despite higher total N (TN) concentrations in agricultural streams (1,520 ± 1,640 vs. 780 ± 600 µg N/L). Although clear patterns linking N2 O concentrations and environmental variables were difficult to discern, the percent saturation of N2 O in the streams was positively correlated with stream concentration of TN and negatively correlated with pH. We speculate that the apparent contradiction between lower TN concentration but similar N2 O concentrations in forest streams than in agricultural streams is due to the low pH (<6) in forest soils and streams which affects denitrification and yields higher N2 O emissions. An estimate of the N2 O emission from low-order streams at the national scale revealed that ~1.8 × 109  g N2 O-N are emitted annually in Sweden, with forest streams contributing about 80% of the total stream emission. Hence, our results provide evidence that forest streams can act as substantial N2 O sources in the landscape with 800 × 109  g CO2 -eq emitted annually in Sweden, equivalent to 25% of the total N2 O emissions from the Swedish agricultural sector.

Nanoparticles transported from aquatic to terrestrial ecosystems via emerging aquatic insects compromise subsidy quality.

Nanoparticle contaminants enter aquatic ecosystems and are transported along the stream network. Here, we demonstrate a novel pathway for the return of nanoparticles from aquatic to terrestrial ecosystems via cross-boundary subsidies. During their emergence, trichopteran caddisflies carried titanium dioxide and gold nanoparticles into their terrestrial life stages. Moreover, their emergence was delayed by ≤30 days, and their energy reserves were depleted by ≤25%. Based on worst case estimates, it is suggested that terrestrial predators, such as bats feeding on aquatic prey, may ingest up to three orders of magnitude higher gold levels than anticipated for humans. Additionally, terrestrial predator species may suffer from alterations in the temporal availability and nutritional quality of their prey. Considering the substantial transfer of insect biomass to terrestrial ecosystems, nanoparticles may decouple aquatic and terrestrial food webs with important (meta-)ecosystem level consequences.

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.

A blessing in disguise? Natural organic matter reduces the UV light-induced toxicity of nanoparticulate titanium dioxide.

Besides their economic value, engineered inorganic nanoparticles (EINPs) may pose a risk for the integrity of ecosystems. Among EINPs, titanium dioxide (nTiO2) is frequently used and released into surface waters in the µg range. There, nTiO2 interacts with environmental factors, influencing its potential to cause adverse effects on aquatic life. Although factors like ultra violet (UV) light and natural organic matter (NOM) are considered as ubiquitous, their joint impact on nTiO2-induced toxicity is poorly understood. This study addressed the acute toxicity of nTiO2 (P25; 0.00-64.00 mg/L; ~60 nm) at ambient UV light (0.00-5.20 W UVA/m2) and NOM levels (seaweed extract; 0.00-4.00 mg TOC/L), using the immobility of Daphnia magna as response variable. Confirming previous studies, effects caused by nTiO2 were elevated with increasing UV radiation (up to ~280 fold) and mitigated by higher NOM levels (up to ~12 fold), possibly due to reduced reactive oxygen species (ROS; measured as •OH radicals) formation at lower UV intensities. However, contradicting to former studies, nTiO2-mediated ROS formation was not proportional to increasing NOM levels: lower concentrations (0.04-0.40 mg TOC/L) slightly diminished, whereas a higher concentration (4.00 mg TOC/L) promoted the ROS quantity, irrespective of UV intensity. Measured ROS levels do not fully explain the observed nTiO2-induced toxicity, whereas increasing acetylcholinesterase and glutathione-S-transferase activities in daphnids (in presence of 8.00 mg/L nTiO2 and elevated UV intensity) point towards neurotoxic and oxidative stress as a driver for the observed effects. Hence, despite higher •OH levels in the treatments where 4.00 mg TOC/L were present, NOM was still capable of reducing nTiO2-induced stress and ultimately adverse effects in aquatic life.

Similar recovery time of microbial functions from fungicide stress across biogeographical regions.

Determining whether the structural and functional stress responses of communities are similar across space and time is paramount for forecasting and extrapolating the consequences of anthropogenic pressures on ecosystems and their services. Stream ecosystems are under high anthropogenic pressure; however, studies have only examined the response of stream communities across large scales over multiple generations. We studied the responses of leaf-associated microbial communities in streams within three European biogeographical regions to chemical stress in a microcosm experiment with multiple cycles of fungicide pollution and resource colonisation. Fungal community composition and the ecosystem function leaf decomposition were measured as response variables. Microbial leaf decomposition showed similar recovery times under environmental levels of fungicide exposure across regions. Initially, the decomposition declined (between 19 and 53%) under fungicide stress and recovered to control levels during the third cycle of pollution and colonisation. Although community composition and its stress response varied between regions, this suggests similar functional community adaptation towards fungicide stress over time. Genetic, epigenetic and physiological adaptations, as well as species turnover, may have contributed to community adaptation but further studies are required to determine if and to which extent these mechanisms are operating. Overall, our findings provide the first evidence of a similar functional response of microbial leaf decomposition to chemical stress across space and time.

SETAC GLB and SETAC Europe SAC: a liaison promoting the next generation of ecotoxicologists and environmental chemists.

This commentary is an introduction for students to the Society of Environmental Toxicology and Chemistry (SETAC) and its Student Advisory Council (SAC). As young academics face challenges while trying to develop their careers, SETAC and the SAC help facilitate student involvement in the various communities within the society that can help to develop the students' careers within the environmental sciences [e.g. the German Language Branch (GLB)]. This piece would also like to emphasize and pay homage to the continual cooperation between the SAC and the ESEU, which provides a scientific platform to communicate internationally and beyond the borders of SETAC, as well as offer heartfelt congratulations from the SAC to the GLB for their "20 Years SETAC GLB" and deep gratitude for their strong advocacy and support of the SAC.

Blinded by the light: Increased chlorophyll fluorescence of herbicide-exposed periphyton masks unfavorable structural responses during exposure and recovery.

In surface waters within agricultural catchments, periphyton - i.e., biofilms containing algae, heterotrophs, and associated detritus - is subjected to multiple stressors including herbicides. Although herbicide effects on periphyton are frequently studied, the focus has been on photosynthesis-inhibiting herbicides while other modes of toxic action have received little attention. Against this background, a 21-days-lasting bioassay was conducted, during which mature periphytic communities were exposed to the carotenoid-biosynthesis-inhibiting herbicide diflufenican for 12 days (up to 10 µg/L; n = 4), followed by a 9-days-lasting recovery phase in herbicide-free medium. Variables related to periphytic functioning (photosynthetic efficiency and non-photochemical quenching) and structure (pigment concentrations, biomass, and algal community structure) were quantified every third day during both experimental phases. Exposure to ≥ 0.2 µg diflufenican/L resulted in 20-25% and 25-30% lowered carotenoid and chlorophyll a concentrations, respectively, likely explained by a reduced algal biovolume as well as diflufenican's mode of toxic action and thus a shift towards a higher heterotrophy of the communities. Despite these adverse effects on the photosynthetic apparatus, the photosynthetic efficiency increased by up to ∼15% under diflufenican exposure judged on higher chlorophyll fluorescence. This may be explained by an up to ∼60% reduced non-photochemical quenching as well as binding of diflufenican to the pigment-protein membrane complex of the photosystem II, two processes causing higher chlorophyll fluorescence. Additionally, phototrophs may have actively increased energy assimilation to cope with higher energy demands under chemical stress. Although periphyton showed some recovery potential following the exposure phase, observed as increasing chlorophyll a concentrations and non-photochemical quenching, periphyton may not be able to quickly recover from stress given the persistent increase in the photosynthetic efficiency. While the processes underlying the observed effects yet remain speculative, the results suggest a shift towards a higher degree of heterotrophy in periphytic communities ultimately increasing the importance of heterotrophic ecosystem functions at impacted sites over the long term.

Assessing the effects of field-relevant pesticide mixtures for their compliance with the concentration addition model - An experimental approach with Daphnia magna.

The environmental risk assessment of pesticides is mainly performed on individual active ingredients. In surface waters within the agricultural landscape, however, contamination is usually characterized by complex pesticide mixtures. To estimate the joint effects caused by these complex mixtures, mathematical models have been proposed. Among these, the model of concentration addition (CA) is suggested as default model for the risk assessment of chemical mixtures as it is considered protective for mixtures composed of similar and dissimilar acting substances. Here we assessed the suitability of CA predictions for seven field relevant pesticide mixtures using acute (immobility) and chronic (reproduction) responses of the standard test species Daphnia magna. Pesticide mixtures indicated largely additive or less than additive effects when using CA model predictions as a reference. Moreover, we revealed that deviations from CA predictions are lower for chronic (up to 3.2-fold) relative to acute (up to 7.2-fold) response variables. Additionally, CA predictions were in general more accurate for complex mixtures relative to those composed of only a few pesticides. Thus, this study suggests CA models as largely protective for the risk assessment of pesticide mixtures justifying its use as default model. At the same time, extrapolating conclusions about the joint effects of pesticides from acute to chronic responses is uncertain, due to partly large discrepancies with regards to the deviation of model prediction and observed effects between exposure scenarios.

The evil within? Systemic fungicide application in trees enhances litter quality for an aquatic decomposer-detritivore system.

Waterborne exposure towards fungicides is known to trigger negative effects in aquatic leaf-associated microbial decomposers and leaf-shredding macroinvertebrates. We expected similar effects when these organisms use leaf material from terrestrial plants that were treated with systemic fungicides as a food source since the fungicides may remain within the leaves when entering aquatic systems. To test this hypothesis, we treated black alder (Alnus glutinosa) trees with a tap water control or a systemic fungicide mixture (azoxystrobin, cyprodinil, quinoxyfen, and tebuconazole) at two worst-case application rates. Leaves of these trees were used in an experiment targeting alterations in two functions provided by leaf-associated microorganisms, namely the decomposition and conditioning of leaf material. The latter was addressed via the food-choice response of the amphipod shredder Gammarus fossarum. During a second experiment, the potential impact of long-term consumption of leaves from trees treated with systemic fungicides on G. fossarum was assessed. Systemic fungicide treatment altered the resource quality of the leaf material resulting in trends of increased fungal spore production and an altered community composition of leaf-associated fungi. These changes in turn caused a significant preference of Gammarus for microbially conditioned leaves that had received the highest fungicide treatment over control leaves. This higher food quality ultimately resulted in a higher gammarid growth (up to 300% increase) during the long-term feeding assay. Although the underlying mechanisms still need to be addressed, the present study demonstrates a positive indirect response in aquatic organisms due to systemic pesticide application in a terrestrial system. As the effects from the introduction of plant material treated with systemic fungicides strongly differ from those mediated via other pathways (e.g., waterborne exposure), our study provides a novel perspective of fungicide-triggered effects in aquatic detritus-based food webs.