Resource-dependent foraging behaviour of grazers enhances effects of nutrient enrichment on algal biomass.

Both the quantity and nutritional quality of food resources can strongly influence the foraging movements of herbivores, which in turn determine the strength of top-down control on primary producer biomass. Nutrient enrichment can alter the biomass and nutritional quality of primary producers, but the consequences for the foraging of herbivores and hence for top-down control are still poorly understood. In this study, we combined a two-factorial experiment (two nutrient levels × grazing by the freshwater gastropod Ancylus fluviatilis) with video analyses tracking grazers' movements to investigate nutrient enrichment effects on spatial ranges of grazing activity and algal biomass removal. Natural stream biofilms were grown in phosphorus-enriched (P+) and phosphorus-poor flumes (P-) for two weeks before A. fluviatilis were added to the flumes and allowed to graze on biofilm for an additional 2 weeks. Total periphyton biomass was enhanced by P+ and reduced by grazer presence. However, the total grazer effect depended on the nutrient level: at the end of the experiment, on average 95% of algal cover were removed by grazing in the P- flumes versus 26% in the P+ flumes. Fast movements of A. fluviatilis were detected significantly more often in the P- treatment, whereas grazers were detected resting more often in the P+ treatment. Our results demonstrate that nutrient enrichment can increase primary producer biomass both directly and indirectly by limiting the foraging ranges of herbivores. The resulting feedback loop between reduced grazing activity and increased plant biomass might in turn exacerbate eutrophication effects on habitat structure.

Taxonomic Shift Over a Phosphorus Gradient Affects the Stoichiometry and Fatty Acid Composition of Stream Periphyton.

Phosphorus enrichment of stream ecosystems generally increases primary production in the benthos, but the consequences of eutrophication for the nutritional quality of periphyton for grazers are less clear. On short timescales, high phosphorus inputs may lead to reduced C:P ratios and high essential fatty acid contents of periphyton, which are both considered important determinants of food quality for grazers. However, nutrient enrichment may alter the taxonomic composition of periphyton and favor the growth of less palatable algal taxa. In this study, periphyton was grown under a gradient of dissolved phosphorus availability from 5 to 100 µg P · L-1 , to investigate eutrophication effects on periphyton taxonomy, C:N:P stoichiometry, and fatty acid composition. After 1 month, periphyton grown under oligotrophic conditions was mainly composed of diatoms (~86%). With increasing phosphorus availability, diatoms were gradually outcompeted by chlorophytes and cyanobacteria, which were the predominant taxon under eutrophic conditions. Unexpectedly, periphyton C:P ratios increased with greater phosphorus supply, from ~280 under oligotrophic conditions up to ~790 at 100 µg · L-1 , reflecting a tendency of chlorophytes and cyanobacteria to produce more biomass per unit of assimilated phosphorus compared to diatoms. Periphyton content of essential polyunsaturated fatty acids relative to biomass followed a unimodal relationship with phosphorus availability and peaked at intermediate phosphorus levels, likely as a result of both taxonomic and nutrient effects. Our results demonstrate that phosphorus-driven eutrophication of freshwater ecosystems may worsen periphyton nutritional quality due to taxonomic sorting, which may further lead to lower growth and reproduction of herbivores.

Continuous In-Stream Assimilatory Nitrate Uptake from High-Frequency Sensor Measurements.

Recently developed in situ NO3­ sensors provide new opportunities to measure changes in stream concentration at high temporal frequencies that historically have not been feasible. In this study, we used multiparameter sensor measurements to relate assimilatory NO3­ uptake to metabolic rates and calculate continuous uptake rates for two stream reaches and a whole stream network. Two years of continuous 15 min data from a forest and agricultural reach of the Selke river (456 km2) revealed a strong correlation between assimilatory NO3­ uptake and growth primary production (GPP) for the forest (r2 = 0.72) and agricultural (r2 = 0.56) stream reach. The slopes of these regressions were in agreement with predicted assimilatory N-uptake based on additional metabolic data. Mean yearly assimilatory NO3­ uptake rates (Ua) were 7.4 times higher in the agricultural stream reach (mean 78.3 mg N m­2 d­1, max 270 mg N m­2 d­1) than in the forest stream reach (mean 10.7 mg N m­2 d­1, max 97.5 mg N m­2 d­1). Nitrate uptake velocities (Vf,a) tended to decrease with increasing nitrate concentrations for periods with high light availability. Percentage daily assimilatory NO3­ uptake peaked at 47.4% of the daily NO3­ loading input to the stream network across the entire watershed, whereas the percentage yearly assimilatory NO3­ uptake was 9.0% of nitrogen loading to the stream network. This is a maximum because uptake can be revered by mineralization processes. The percentage yearly assimilatory NO3­ uptake was lower in the forest-dominated upstream subwatershed (4.8%) than in the lower agriculture dominated subwatershed (13.4%).

Relationship between the elemental composition of stream biofilms and water chemistry-a catchment approach.

As benthic biofilms mediate essential functions in stream ecosystems (e.g., carbon flux, storage of nutrients and other substances), the element-specific regulation of the biofilm composition is of great interest. We tested whether (1) the elemental composition of biofilms is related to that of the water column and (2) there are different accumulation patterns from the dissolved phase (adsorption) and the particulate phase (incorporation of suspended matter). We analysed biomass parameters, nutrients and metals in biofilms and surface waters at 28 sites within a stream network (Bode catchment, Germany). Algal biomass in biofilms was dominated by diatoms. The P/C ratio in biofilms was positively related to total phosphorus of surface water (and to the proportion of agricultural area in the catchment) indicating phosphorus limitation of biofilms, whereas the N/C ratio was not related to nitrate levels of surface water, and neither the P/C nor the N/C ratio to the concentration of dissolved organic carbon (DOC) of surface water. Biofilms were enriched in metals compared to their concentrations in water. The metals in biofilms were positively related to the concentration of dissolved metals in surface water for iron and strontium (but not for manganese, copper, zinc, arsenic or lead) and to the concentrations of particle-associated metals of surface waters for strontium and lead. Manganese and arsenic were the metals with a negative effect on the biomasses of biofilm diatoms and cyanobacteria. Overall, we observed element-specific accumulation patterns in biofilms with selected elements being related to the water column while others were probably subject to biofilm-internal processes.

Saturation of intracellular phosphorus uptake and prevalence of extracellular phosphorus entrapment in fluvial biofilms after long-term P pulses: Implications for river self-purification.

Microbial consortia in riverbed substrates and their extracellular matrix (biofilms) play a key role in phosphorus (P) entrapment. When P entrapment saturates, the benthic compartment changes from a P sink to a P source thus increasing eutrophication risk. P entrapment saturation is expected to differ between intracellular and extracellular P entrapment and between different magnitudes and durations of P inputs. We studied biofilm P-entrapment following short (48 h) and long (14 days) P loading events in stream bypass flumes supplied with a gradient of dissolved P concentrations. This allowed us to link local biofilm processes in sediments to potential effects on river self-purification, via quantifying the P removal efficiency in the flumes. We found that in short-term events, biofilms develop intracellular mechanisms to cope with P inputs, while long-term events and high P inputs suppress the intracellular uptake mechanisms and increase the prevalence of extracellular entrapment. Specifically, long-term events lowered the threshold for intracellular P entrapment saturation, and decreased the ratio between intracellular and extracellular entrapment resulting in lower removal efficiency for dissolved phosphorus. Our results highlight the risk that aquatic ecosystems may face as the ratio of intracellular to extracellular P entrapment decreases, which may reduce their ability to deal with P inputs, thereby increasing risks of eutrophication.

Long-term recovery of benthic food webs after stream restoration.

The assessment of restoration success often neglects trophic interactions within food webs, focusing instead on biodiversity and community structure. Here, we analysed the long-term recovery of food web structure based on stable isotopes (δ13C and δ15N) of benthic invertebrates and quantified responses of food web metrics to time since restoration. The samples derived from twelve restored sites with different restoration ages, sampled annually from 2012 to 2021, and covering an investigation period of up to 28 years after restoration for the whole catchment. Temporal developments of the restored sites were compared to the development of two near-natural sites. The restoration measures consisted of the cessation of sewage inflow and morphological restoration of the channels. As a clear and consistent result over almost all sites, trophic similarity (proportion of co-existing species occupying similar trophic niches) increased with time since restoration, and reached values of near-natural sites, suggesting an increase in the stability and resilience of the food webs. Surprisingly, resource diversity decreased at most restored sites within 10 years after restoration, probably due to the removal of wastewater-derived resources, and a shift towards leaf litter as the dominant resource following the regrowth of the riparian vegetation. Food chain length showed no consistent pattern over time at the different sites both increasing and decreasing with time since restoration. Overall, restoration had clear effects on the food web structure of stream ecosystems. While some effects such as the increase in trophic similarity were consistent at almost all sites, others such as response of the food chain length were context dependent. The study demonstrates the potential of utilizing food web metrics, particularly trophic similarity, in restoration research to achieve a more holistic understanding of ecosystem recovery.

Contribution of groundwater-borne nutrients to eutrophication potential and the share of benthic algae in a large lowland river.

Groundwater inflow can be a significant source of nutrients for riverine ecosystems, which can affect eutrophication i.e., the elevated primary production and the corresponding accumulation of algal biomass. Experimental and modelling work has shown that benthic algae (autotrophic biofilms) in particular benefit, as they have direct access to the inflowing groundwater-borne nutrients. Primarily the supply of phosphorus (P) enhances pelagic algal biomass, as it is the limiting nutrient for primary production in most freshwater systems. In this study, we estimate the effect of groundwater inflow on overall eutrophication of a large, European lowland river and tested its seasonal effect on biofilms in particular. We calculated the effects on overall eutrophication during summer according to the estimated input of groundwater-borne P and the C:P stoichiometry of planktonic algae in the Elbe River. Our model indicated that these diffuse P inputs have the potential to significantly increase eutrophication. Groundwater-P can contribute up to 1.5 t/d PO4 over the investigated 450 km stretch of the Elbe River under low flow conditions. This would result in an additional planktonic load of about 46 t/d of particulate organic carbon, thereby contributing to eutrophication at the regional scale in this river. In contrast, at the local scale, biofilms were collected seasonally from artificial substrata exposed in the river either in hydrogeologically active areas with groundwater inflow, or in areas of varying hydraulic connectivity. Analyses of biofilm macronutrients, structural components and biofilm community composition show distinct effects of season, hydrogeology and groundwater inflow. The dominant predictors were season and the interaction between hydrogeology and groundwater. Benthic eutrophication is most likely to occur in autumn in areas of loose rock with high groundwater inflow. The strong interaction of environmental factors in determining benthic eutrophication highlights the need to assess these factors in combination rather than in isolation.

Linking the evolution of habitat choice to ecosystem functioning: direct and indirect effects of pond-reproducing fire salamanders on aquatic-terrestrial subsidies.

Shifts in life history traits and in the behaviour of species can potentially alter ecosystem functioning. The reproduction of the central European fire salamander (Salamandra salamandra), which usually deposits its larvae in first-order streams, in small pool and pond-like habitats, is an example of a recent local adaptation in this species. Here we aimed to quantify the direct and indirect effects of the predatory larvae on the aquatic food webs in the ponds and on the flux of matter between the ponds and adjacent terrestrial habitats. Our estimates are based on biomass data of the present pond fauna as well as on the analysis of stomach content data, growth rates and population dynamics of the salamander larvae in pond habitats. By their deposition of larvae in early spring, female fire salamanders import between 0.07 and 2.86 g dry mass m(-2) larval biomass into the ponds. Due to high mortality rates in the larval phase and the relatively small size at metamorphosis of the pond-adapted salamanders compared to stream-adapted ones, the biomass export of the metamorphosed salamanders clearly falls below the initial biomass import. Catastrophic events such as high water temperatures and low oxygen levels may even occasionally result in mass mortalities of salamander larvae and thus in a net 100 % import of the salamander biomass into the pond food webs. Indirect effects further accelerate this net import of matter into the aquatic habitat, e.g. the feeding of salamanders on aquatic insect larvae with the emergence of terrestrial adults-thus preventing export-and on terrestrial organisms that fall on the water surface (supporting import). This study demonstrates that the adaptation of salamanders to pond reproduction can alter food web linkages across ecosystem boundaries by enhancing the flux of materials and energy from terrestrial (i.e. forest) to the aquatic (i.e. pond) habitat.

Large-scale patterns in biodiversity of microbial eukaryotes from the abyssal sea floor.

Eukaryotic microbial life at abyssal depths remains "uncharted territory" in eukaryotic microbiology. No phylogenetic surveys have focused on the largest benthic environment on this planet, the abyssal plains. Moreover, knowledge of the spatial patterns of deep-sea community structure is scanty, and what little is known originates primarily from morphology-based studies of foraminiferans. Here we report on the great phylogenetic diversity of microbial eukaryotic communities of all 3 abyssal plains of the southeastern Atlantic Ocean--the Angola, Cape, and Guinea Abyssal Plains--from depths of 5,000 m. A high percentage of retrieved clones had no close representatives in genetic databases. Many clones were affiliated with parasitic species. Furthermore, differences between the communities of the Cape Abyssal Plain and the other 2 abyssal plains point to environmental gradients apparently shaping community structure at the landscape level. On a regional scale, local species diversity showed much less variation. Our study provides insight into the community composition of microbial eukaryotes on larger scales from the wide abyssal sea floor realm and marks a direction for more detailed future studies aimed at improving our understanding of deep-sea microbes at the community and ecosystem levels, as well as the ecological principles at play.

Biogeochemical patterns in a river network along a land use gradient.

The Bode catchment (Germany) shows strong land use gradients from forested parts of the National Park (23% of total land cover) to agricultural (70%) and urbanised areas (7%). It is part of the Terrestrial Environmental Observatories of the German Helmholtz association. We performed a biogeochemical analysis of the entire river network. Surface water was sampled at 21 headwaters and at ten downstream sites, before (in early spring) and during the growing season (in late summer). Many parameters showed lower concentrations in headwaters than in downstream reaches, among them nutrients (ammonium, nitrate and phosphorus), dissolved copper and seston dry mass. Nitrate and phosphorus concentrations were positively related to the proportion of agricultural area within the catchment. Punctual anthropogenic loads affected some parameters such as chloride and arsenic. Chlorophyll a concentration and total phosphorus in surface waters were positively related. The concentration of dissolved organic carbon (DOC) was higher in summer than in spring, whereas the molecular size of DOC was lower in summer. The specific UV absorption at 254 nm, indicating the content of humic substances, was higher in headwaters than in downstream reaches and was positively related to the proportion of forest within the catchment. CO2 oversaturation of the water was higher downstream compared with headwaters and was higher in summer than in spring. It was correlated negatively with oxygen saturation and positively with DOC concentration but negatively with DOC quality (molecular size and humic content). A principle component analysis clearly separated the effects of site (44%) and season (15%), demonstrating the strong effect of land use on biogeochemical parameters.

Shifts from cooperative to individual-based predation defense determine microbial predator-prey dynamics.

Predation defense is an important feature of predator-prey interactions adding complexity to ecosystem dynamics. Prey organisms have developed various strategies to escape predation which differ in mode (elude vs. attack), reversibility (inducible vs. permanent), and scope (individual vs. cooperative defenses). While the mechanisms and controls of many singular defenses are well understood, important ecological and evolutionary facets impacting long-term predator-prey dynamics remain underexplored. This pertains especially to trade-offs and interactions between alternative defenses occurring in prey populations evolving under predation pressure. Here, we explored the dynamics of a microbial predator-prey system consisting of bacterivorous flagellates (Poteriospumella lacustris) feeding on Pseudomonas putida. Within five weeks of co-cultivation corresponding to about 35 predator generations, we observed a consistent succession of bacterial defenses in all replicates (n = 16). Initially, bacteria expressed a highly effective cooperative defense based on toxic metabolites, which brought predators close to extinction. This initial strategy, however, was consistently superseded by a second mechanism of predation defense emerging via de novo mutations. Combining experiments with mathematical modeling, we demonstrate how this succession of defenses is driven by the maximization of individual rather than population benefits, highlighting the role of rapid evolution in the breakdown of social cooperation.

Large-scale nutrient and carbon dynamics along the river-estuary-ocean continuum.

Nutrient and carbon dynamics within the river-estuary-coastal water systems are key processes in understanding the flux of matter from the terrestrial environment to the ocean. Here, we analysed those dynamics by following a sampling approach based on the travel time of water and an advanced calculation of nutrient fluxes in the tidal part. We started with a nearly Lagrangian sampling of the river (River Elbe, Germany; 580 km within 8 days). After a subsequent investigation of the estuary, we followed the plume of the river by raster sampling the German Bight (North Sea) using three ships simultaneously. In the river, we detected intensive longitudinal growth of phytoplankton connected with high oxygen saturation and pH values and an undersaturation of CO2, whereas concentrations of dissolved nutrients declined. In the estuary, the Elbe shifted from an autotrophic to a heterotrophic system: Phytoplankton died off upstream of the salinity gradient, causing minima in oxygen saturation and pH, supersaturation of CO2, and a release of nutrients. In the shelf region, phytoplankton and nutrient concentrations were low, oxygen was close to saturation, and pH was within a typical marine range. Over all sections, oxygen saturation was positively related to pH and negatively to pCO2. Corresponding to the significant particulated nutrient flux via phytoplankton, flux rates of dissolved nutrients from river into estuary were low and determined by depleted concentrations. In contrast, fluxes from the estuary to the coastal waters were higher and the pattern was determined by tidal current. Overall, the approach is appropriate to better understand land-ocean fluxes, particularly to illuminate the importance of these fluxes under different seasonal and hydrological conditions, including flood and drought events.

Seasonal and successional influences on bacterial community composition exceed that of protozoan grazing in river biofilms.

The effects of protozoa (heterotrophic flagellates and ciliates) on the morphology and community composition of bacterial biofilms were tested under natural background conditions by applying size fractionation in a river bypass system. Confocal laser scanning microscopy (CLSM) was used to monitor the morphological structure of the biofilm, and fingerprinting methods (single-stranded conformation polymorphism [SSCP] and denaturing gradient gel electrophoresis [DGGE]) were utilized to assess changes in bacterial community composition. Season and internal population dynamics had a greater influence on the bacterial biofilm than the presence of protozoa. Within this general framework, bacterial area coverage and microcolony abundance were nevertheless enhanced by the presence of ciliates (but not by the presence of flagellates). We also found that the richness of bacterial operational taxonomic units was much higher in planktonic founder communities than in the ones establishing the biofilm. Within the first 2 h of colonization of an empty substrate by bacteria, the presence of flagellates additionally altered their biofilm community composition. As the biofilms matured, the number of bacterial operational taxonomic units increased when flagellates were present in high abundances. The additional presence of ciliates tended to at first reduce (days 2 to 7) and later increase (days 14 to 29) bacterial operational taxonomic unit richness. Altogether, the response of the bacterial community to protozoan grazing pressure was small compared to that reported in planktonic studies, but our findings contradict the assumption of a general grazing resistance of bacterial biofilms toward protozoa.

Demonstration of an aggregated biomarker response approach to assess the impact of point and diffuse contaminant sources in feral fish in a small river case study.

The assessment of the exposure of aquatic wildlife to complex environmental mixtures of chemicals originating from both point and diffuse sources and evaluating the potential impact thereof constitutes a significant step towards mitigating toxic pressure and the improvement of ecological status. In the current proof-of-concept study, we demonstrate the potential of a novel Aggregated Biomarker Response (ABR) approach involving a comprehensive set of biomarkers to identify complex exposure and impacts on wild brown trout (Salmo trutta fario). Our scenario used a small lowland river in Germany (Holtemme river in the Elbe river catchment) impacted by two wastewater treatment plants (WWTP) and diffuse agricultural runoff as a case study. The trout were collected along a pollution gradient (characterised in a parallel study) in the river. Compared to fish from the reference site upstream of the first WWTP, the trout collected downstream of the WWTPs showed a significant increase in micronucleus formation, phase I and II enzyme activities, and oxidative stress parameters in agreement with increasing exposure to various chemicals. By integrating single biomarker responses into an aggregated biomarker response, the two WWTPs' contribution to the observed toxicity could be clearly differentiated. The ABR results were supported by chemical analyses and whole transcriptome data, which revealed alterations of steroid biosynthesis and associated pathways, including an anti-androgenic effect, as some of the key drivers of the observed toxicity. Overall, this combined approach of in situ biomarker responses complemented with molecular pathway analysis allowed for a comprehensive ecotoxicological assessment of fish along the river. This study provides evidence for specific hazard potentials caused by mixtures of agricultural and WWTP derived chemicals at sublethal concentrations. Using aggregated biomarker responses combined with chemical analyses enabled an evidence-based ranking of sites with different degrees of pollution according to toxic stress and observed effects.

Lagrangian profiles of riverine autotrophy, organic matter transformation, and micropollutants at extreme drought.

On their way from inland to the ocean, flowing water bodies, their constituents and their biotic communities are exposed to complex transport and transformation processes. However, detailed process knowledge as revealed by Lagrangian measurements adjusted to travel time is rare in large rivers, in particular at hydrological extremes. To fill this gap, we investigated autotrophic processes, heterotrophic carbon utilization, and micropollutant concentrations applying a Lagrangian sampling design in a 600 km section of the River Elbe (Germany) at historically low discharge. Under base flow conditions, we expect the maximum intensity of instream processes and of point source impacts. Phytoplankton biomass and photosynthesis increased from upstream to downstream sites but maximum chlorophyll concentration was lower than at mean discharge. Concentrations of dissolved macronutrients decreased to almost complete phosphate depletion and low nitrate values. The longitudinal increase of bacterial abundance and production was less pronounced than in wetter years and bacterial community composition changed downstream. Molecular analyses revealed a longitudinal increase of many DOM components due to microbial production, whereas saturated lipid-like DOM, unsaturated aromatics and polyphenols, and some CHOS surfactants declined. In decomposition experiments, DOM components with high O/C ratios and high masses decreased whereas those with low O/C ratios, low masses, and high nitrogen content increased at all sites. Radiocarbon age analyses showed that DOC was relatively old (890-1870 years B.P.), whereas the mineralized fraction was much younger suggesting predominant oxidation of algal lysis products and exudates particularly at downstream sites. Micropollutants determining toxicity for algae (terbuthylazine, terbutryn, isoproturon and lenacil), hexachlorocyclohexanes and DDTs showed higher concentrations from the middle towards the downstream part but calculated toxicity was not negatively correlated to phytoplankton. Overall, autotrophic and heterotrophic process rates and micropollutant concentrations increased from up- to downstream reaches, but their magnitudes were not distinctly different to conditions at medium discharges.

WITHDRAWN: Wide distribution ranges of several microbial eukaryotes of nanofaunal size at abyssal depths of the South-East Atlantic and the Mediterranean.

This article has been withdrawn.

Feedback between bottom-up and top-down control of stream biofilm mediated through eutrophication effects on grazer growth.

Algal biofilms in streams are simultaneously controlled by light and nutrient availability (bottom-up control) and by grazing activity (top-down control). In addition to promoting algal growth, light and nutrients also determine the nutritional quality of algae for grazers. While short-term experiments have shown that grazers increase consumption rates of nutrient-poor algae due to compensatory feeding, nutrient limitation in the long run can constrain grazer growth and hence limit the strength of grazing activity. In this study, we tested the effects of light and phosphorus availability on grazer growth and thus on the long-term control of algal biomass. At the end of the experiment, algal biomass was significantly affected by light, phosphorus and grazing, but the interactive effects of the three factors significantly changed over time. At both high light and phosphorus supply, grazing did not initially reduce algal biomass, but the effect of grazing became stronger in the final three weeks of the experiment. Snail growth was enhanced by light, rather than phosphorus, suggesting that algal quantity rather than quality was the main limiting factor for grazer growth. Our results highlight the role of feedback effects and the importance of long-term experiments in the study of foodweb interactions.

Long-term effects of a catastrophic insecticide spill on stream invertebrates.

Accidental spills or illegal discharges of pesticides in aquatic ecosystems can lead to exposure levels that strongly exceed authorized pesticide concentrations, causing major impacts on aquatic ecosystems. Such short-term events often remain undetected in regular monitoring programs with infrequent sampling. In early spring 2015, we identified a catastrophic pesticide spill with the insecticide cypermethrin in the Holtemme River, Germany. Based on existing pre-event macroinvertebrate community data, we monitored the effects and recovery of the macroinvertebrate community for more than two years after the spill. Strong short-term effects were apparent for all taxa with the exception of Chironomidae and Tubificidae. Effects could also be observed on the community level as total abundance, taxa number and biomass strongly decreased. Total abundance and taxa number showed a fast recovery. Regarding long-term effects, the total biomass remained substantially below the pre-contamination level (76%) until the end of the study. Also the abundances of three taxa (Gammarus, Leuctra, Limnius Ad.) did not return to levels prior to the spill even after 26 months. This lack of the taxon-specific recovery was likely due to their long generation time and a low migration ability due to a restricted connectivity between the contaminated site and uncontaminated stream sections. These factors proved to be stronger predictors for the recovery than the pesticide tolerance. We revealed that the biological indicators SPEARpesticides and share of Ephemeroptera, Plecoptera and Trichoptera (EPT) are not suitable for the identification of such extreme events, when nearly all taxa are eradicated. Both indicators are functioning only when repeated stressors initiate long-term competitive replacement of sensitive by insensitive taxa. We conclude that pesticide spills can have significant long-term effects on stream macroinvertebrate communities. Regular ecological monitoring is imperative to identify such ecosystem impairments, combined with analytical chemistry methods to identify the potential sources of spills.

Pesticides are the dominant stressors for vulnerable insects in lowland streams.

Despite elaborate regulation of agricultural pesticides, their occurrence in non-target areas has been linked to adverse ecological effects on insects in several field investigations. Their quantitative role in contributing to the biodiversity crisis is, however, still not known. In a large-scale study across 101 sites of small lowland streams in Central Europe, Germany we revealed that 83% of agricultural streams did not meet the pesticide-related ecological targets. For the first time we identified that agricultural nonpoint-source pesticide pollution was the major driver in reducing vulnerable insect populations in aquatic invertebrate communities, exceeding the relevance of other anthropogenic stressors such as poor hydro-morphological structure and nutrients. We identified that the current authorisation of pesticides, which aims to prevent unacceptable adverse effects, underestimates the actual ecological risk as (i) measured pesticide concentrations exceeded current regulatory acceptable concentrations in 81% of the agricultural streams investigated, (ii) for several pesticides the inertia of the authorisation process impedes the incorporation of new scientific knowledge and (iii) existing thresholds of invertebrate toxicity drivers are not protective by a factor of 5.3 to 40. To provide adequate environmental quality objectives, the authorisation process needs to include monitoring-derived information on pesticide effects at the ecosystem level. Here, we derive such thresholds that ensure a protection of the invertebrate stream community.

Disentangling multiple chemical and non-chemical stressors in a lotic ecosystem using a longitudinal approach.

Meeting ecological and water quality standards in lotic ecosystems is often failed due to multiple stressors. However, disentangling stressor effects and identifying relevant stressor-effect-relationships in complex environmental settings remain major challenges. By combining state-of-the-art methods from ecotoxicology and aquatic ecosystem analysis, we aimed here to disentangle the effects of multiple chemical and non-chemical stressors along a longitudinal land use gradient in a third-order river in Germany. We distinguished and evaluated four dominant stressor categories along this gradient: (1) Hydromorphological alterations: Flow diversity and substrate diversity correlated with the EU-Water Framework Directive based indicators for the quality element macroinvertebrates, which deteriorated at the transition from near-natural reference sites to urban sites. (2) Elevated nutrient levels and eutrophication: Low to moderate nutrient concentrations together with complete canopy cover at the reference sites correlated with low densities of benthic algae (biofilms). We found no more systematic relation of algal density with nutrient concentrations at the downstream sites, suggesting that limiting concentrations are exceeded already at moderate nutrient concentrations and reduced shading by riparian vegetation. (3) Elevated organic matter levels: Wastewater treatment plants (WWTP) and stormwater drainage systems were the primary sources of bioavailable dissolved organic carbon. Consequently, planktonic bacterial production and especially extracellular enzyme activity increased downstream of those effluents showing local peaks. (4) Micropollutants and toxicity-related stress: WWTPs were the predominant source of toxic stress, resulting in a rapid increase of the toxicity for invertebrates and algae with only one order of magnitude below the acute toxic levels. This toxicity correlates negatively with the contribution of invertebrate species being sensitive towards pesticides (SPEARpesticides index), probably contributing to the loss of biodiversity recorded in response to WWTP effluents. Our longitudinal approach highlights the potential of coordinated community efforts in supplementing established monitoring methods to tackle the complex phenomenon of multiple stress.