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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

Exploring flow-biofilm-sediment interactions: Assessment of current status and future challenges.

Biofilm activities and their interactions with physical, chemical and biological processes are of great importance for a variety of ecosystem functions, impacting hydrogeomorphology, water quality and aquatic ecosystem health. Effective management of water bodies requires advancing our understanding of how flow influences biofilm-bound sediment and ecosystem processes and vice-versa. However, research on this triangle of flow-biofilm-sediment is still at its infancy. In this Review, we summarize the current state of the art and methodological approaches in the flow-biofilm-sediment research with an emphasis on biostabilization and fine sediment dynamics mainly in the benthic zone of lotic and lentic environments. Example studies of this three-way interaction across a range of spatial scales from cell (nm - µm) to patch scale (mm - dm) are highlighted in view of the urgent need for interdisciplinary approaches. As a contribution to the review, we combine a literature survey with results of a pilot experiment that was conducted in the framework of a joint workshop to explore the feasibility of asking interdisciplinary questions. Further, within this workshop various observation and measuring approaches were tested and the quality of the achieved results was evaluated individually and in combination. Accordingly, the paper concludes by highlighting the following research challenges to be considered within the forthcoming years in the triangle of flow-biofilm-sediment: i) Establish a collaborative work among hydraulic and sedimentation engineers as well as ecologists to study mutual goals with appropriate methods. Perform realistic experimental studies to test hypotheses on flow-biofilm-sediment interactions as well as structural and mechanical characteristics of the bed. ii) Consider spatially varying characteristics of flow at the sediment-water interface. Utilize combinations of microsensors and non-intrusive optical methods, such as particle image velocimetry and laser scanner to elucidate the mechanism behind biofilm growth as well as mass and momentum flux exchanges between biofilm and water. Use molecular approaches (DNA, pigments, staining, microscopy) for sophisticated community analyses. Link varying flow regimes to microbial communities (and processes) and fine sediment properties to explore the role of key microbial players and functions in enhancing sediment stability (biostabilization). iii) Link laboratory-scale observations to larger scales relevant for management of water bodies. Conduct field experiments to better understand the complex effects of variable flow and sediment regimes on biostabilization. Employ scalable and informative observation techniques (e.g., hyperspectral imaging, particle tracking) that can support predictions on the functional aspects, such as metabolic activity, bed stability, nutrient fluxes under variable regimes of flow-biofilm-sediment.

Molecular change of dissolved organic matter and patterns of bacterial activity in a stream along a land-use gradient.

Fluvial networks are globally relevant for the processing of dissolved organic matter (DOM). To investigate the change in molecular DOM diversity along the river course, high-field FTICR mass spectrometry and NMR spectroscopy of riverine DOM as well as bacterial abundance and activity were measured in a third order stream along a land-use gradient from pristine, agricultural to urban landscapes. DOM composition showed a clear evolution along the river course with an initial decrease of average oxidation and unsaturation followed by an increased relative abundance of CHNO and CHOS compounds introduced by agriculture and waste water, respectively. DOM composition was dominated by rather unsaturated CHO compounds (H/C ≤ 1) in headwaters and by more aliphatic molecules at downstream sites. Oxygenated functional groups shifted from aromatic ethers and hydroxyl groups to aliphatic carboxylic acids and aliphatic hydroxyl groups. This massive dislocation of oxygen significantly increased the diversity of atomic environments in branched aliphatic groups from headwater to downstream DOM. Mass spectra of DOM enabled the detection of compositional relationships to bacterial abundance and activity which was positively related to more aliphatic components (H/C > 1) and negatively related to unsaturated components. FTICR mass and NMR spectra corroborated the initial decline in DOM molecular diversity predicted by the River Continuum Concept (RCC) but demonstrated an anthropogenic increase in the molecular diversity of DOM further downstream. While the high DOM molecular diversity in first order headwater streams was the result of small scale ecosystem plurality, agriculture and waste water treatment introduced many components in the lower reaches. These anthropogenic influences together with massive bacterial oxidation of DOM contributed to a growth of molecular diversity of downstream DOM whose composition and structure differed entirely from those found in pristine headwaters.

Hydrodynamics Alter the Tolerance of Autotrophic Biofilm Communities Toward Herbicides.

Multiple stressors pose potential risk to aquatic ecosystems and are the main reasons for failing ecological quality standards. However, mechanisms how multiple stressors act on aquatic community structure and functioning are poorly understood. This is especially true for two important stressors types, hydrodynamic alterations and toxicants. Here we perform a mesocosm experiment in hydraulic flumes connected as a bypass to a natural stream to test the interactive effects of both factors on natural (inoculated from streams water) biofilms. Biofilms, i.e., the community of autotrophic and heterotrophic microorganisms and their extracellular polymeric substances (EPS) in association with substratum, are key players in stream functioning. We hypothesized (i) that the tolerance of biofilms toward toxicants (the herbicide Prometryn) decreases with increasing hydraulic stress. As EPS is known as an absorber of chemicals, we hypothesize (ii) that the EPS to cell ratio correlates with both hydraulic stress and herbicide tolerance. Tolerance values were derived from concentration-response assays. Both, the herbicide tolerance and the biovolume of the EPS significantly correlated with the turbulent kinetic energy (TKE), while the diversity of diatoms (the dominant group within the stream biofilms) increased with flow velocity. This indicates that the positive effect of TKE on community tolerance was mediated by turbulence-induced changes in the EPS biovolume. This conclusion was supported by a second experiment, showing decreasing effects of the herbicide to a diatom biofilm (Nitzschia palea) with increasing content of artificial EPS. We conclude that increasing hydrodynamic forces in streams result in an increasing tolerance of microbial communities toward chemical pollution by changes in EPS-mediated bioavailability of toxicants.

The role of hydrodynamics in shaping the composition and architecture of epilithic biofilms in fluvial ecosystems.

Previous laboratory and on-site experiments have highlighted the importance of hydrodynamics in shaping biofilm composition and architecture. In how far responses to hydrodynamics can be found in natural flows under the complex interplay of environmental factors is still unknown. In this study we investigated the effect of near streambed turbulence in terms of turbulent kinetic energy (TKE) on the composition and architecture of biofilms matured in two mountainous streams differing in dissolved nutrient concentrations. Over both streams, TKE significantly explained 7% and 8% of the variability in biofilm composition and architecture, respectively. However, effects were more pronounced in the nutrient richer stream, where TKE significantly explained 12% and 3% of the variability in biofilm composition and architecture, respectively. While at lower nutrient concentrations seasonally varying factors such as stoichiometry of dissolved nutrients (N/P ratio) and light were more important and explained 41% and 6% of the variability in biofilm composition and architecture, respectively. Specific biofilm features such as elongated ripples and streamers, which were observed in response to the uniform and unidirectional flow in experimental settings, were not observed. Microbial biovolume and surface area covered by the biofilm canopy increased with TKE, while biofilm thickness and porosity where not affected or decreased. These findings indicate that under natural flows where near bed flow velocities and turbulence intensities fluctuate with time and space, biofilms became more compact. They spread uniformly on the mineral surface as a film of densely packed coccoid cells appearing like cobblestone pavement. The compact growth of biofilms seemed to be advantageous for resisting hydrodynamic shear forces in order to avoid displacement. Thus, near streambed turbulence can be considered as important factor shaping the composition and architecture of biofilms grown under natural flows.

Grazing resistance of bacterial biofilms: a matter of predators' feeding trait.

Biofilm formation in bacteria is considered to be one strategy to avoid protozoan grazing. However, this assumption is largely based on experiments with suspension-feeding protozoans. Here we test the hypothesis that grazing resistance depends on both the grazers' feeding trait and the bacterial phenotype, rather than being a general characteristic of bacterial biofilms. We combined batch experiments with mathematical modelling, considering the bacterium Pseudomonas putida and either a suspension-feeding (i.e. the ciliate Paramecium tetraurelia) or a surface-feeding grazer (i.e. the amoeba Acanthamoeba castellanii). We find that both plankton and biofilm phenotypes were consumed, when exposed to their specialised grazer, whereas the other phenotype remained grazing-resistant. This was consistently shown in two experiments (starting with either only planktonic bacteria or with additional pre-grown biofilms) and matches model predictions. In the experiments, the plankton feeder strongly stimulated the biofilm biomass. This stimulation of the resistant prey phenotype was not predicted by the model and it was not observed for the biofilm feeders, suggesting the existence of additional mechanisms that stimulate biofilm formation besides selective feeding. Overall, our results confirm our hypothesis that grazing resistance is a matter of the grazers' trait (i.e. feeding type) rather than a biofilm-specific property.

A new approach for evaluating transformations of dissolved organic matter (DOM) via high-resolution mass spectrometry and relating it to bacterial activity.

Streams are important sites of transformation of dissolved organic matter (DOM). The molecular characterization of DOM-quality changes requires sophisticated analytical evaluation techniques. The goal of our study was to link molecular DOM transformation with bacterial activity. We measured the degradation of leaf leachate over a gradient of bacterial production obtained by different rates of percolation of sediments in seven experimental flumes on five sampling dates. We developed a new strategy for evaluating molecular formula data sets obtained by ultra-high resolution Fourier transform ion cyclotron resonance mass spectrometry (FTICR MS), in which the time-dependent change of component abundance was fitted by a linear regression model after normalization of mass peak intensities. All components were categorized by calculating the slope (change of percent intensity per day) in each of the seven flumes. These slopes were then related to cumulative bacterial production. The concentration of DOM decreased quickly in all flumes. Bacterial activity was higher in flumes with percolated sediment than in those without percolation, whereas plankton bacterial activity was higher in flumes without percolation or without sediment. There were no differences in molecular-DOM characteristics between flumes, but there were distinct changes over time. Positive slopes, i.e. increasing intensities over time, were found for small molecules (MW < 450 Da) and high O/C ratios, whereas decreasing intensities were observed less often and only for large molecules and low O/C ratios. The positive slopes of produced components showed a positive relationship to bacterial production for small and for oxygen-rich components. The negative slopes of degraded components were negatively related to bacterial production for large and for oxygen-deficient molecules. Overall, the approach provided new insights into the transformation of specific molecular DOM components.

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.