Effects of low flow and co-occurring stressors on structural and functional characteristics of the benthic biofilm in small streams.

Low flow and co-occurring stress is a more and more frequent phenomenon these years in small agricultural streams as a consequence of climate change. In the present study we explored short and longer term structural responses of the stream benthic algae community and biofilm metabolism to multiple stress in small streams applying a semi-experimental approach. We hypothesized that i) a reduction in flow in combination with secondary stress (nutrients and sediments) have immediate effects on the benthic algae community in terms of biomass (chlorophyll a, biovolume), taxonomic and trait (lifeform and size distribution) compositions as well as on metabolism (GPP and CR), and ii) that changes in the benthic algae community persist due to altered environmental settings but that functional redundancy among benthic algae species provides a high level of resilience in metabolism (GPP and CR). Overall, we found that stress imposed by nutrients was less pronounced than stress imposed by fine sediments under low flow, and that nutrient enrichment to some extent mitigated effects of fine sediments. Fine sediment deposition mediated a decline in the fraction of erect algae and/or algae with mucilage stalks but this did not happen under co-occurring stress from both sediments and nutrients. Additionally, fine sediment deposition mediated a decline in GPP of the biofilm, but again this did not happen under co-occurring stress from nutrients. We conclude that 1) the benthic algae community and biofilm metabolism displayed similar resilience to stress imposed by low flow and co-occurring stress from nutrients and sediments on a short and longer time scale and 2) as structure-function adaptations may occur at several trophic levels in the biofilm, more research is needed to explore mechanisms underlying mitigating effects of nutrients in response to sediment deposition under low flow.

The future of European water management: Demonstration of a new WFD compliant framework to support sustainable management under multiple stress.

The Water Framework Directive (WFD), which is the most comprehensive instrument of EU water policy, is more relevant than ever. Sixty percent of Europe's surface water bodies still fail to achieve good ecological status and a multitude of new stressors continue to emerge. A sustained and wholehearted water management effort is therefore of highest priority. Here, we present a new biological assessment approach specifically designed to safeguard sustainable water management under multiple stress. The framework contains three independent elements: 1) an ecological assessment system based on community abundance and composition to quantify ecological status; 2) a diagnostic tool to identify cause(s) of ecological degradation; 3) a management platform to guide the choice of relevant mitigation measures for improvement of the ecological status. The proposed framework is fully compliant with the WFD and currently applied in the assessment of aquatic plant communities in Danish streams. Importantly, the approach presented is not restricted to specific taxonomic groups or ecosystem types but is an example of how a simple approach can bring the conceptual idea of the WFD - that community characteristics in unimpacted, type-specific water bodies should be the backbone in ecological assessments - into practice.

Successful restoration of a tropical shallow eutrophic lake: Strong bottom-up but weak top-down effects recorded.

Fish manipulation has been used to restore lakes in the temperate zone. Often strong short-term cascading effects have been obtained, but the long term-perspectives are less clear. Fish manipulation methods are far less advanced for warm lakes, and it is debatable whether it is, in fact, possible to create a trophic cascade in warm lakes due to the dominance and high densities of fast-reproducing omnivorous fish. However, removal of benthic feeding fish also reduce disturbance of the sediment, which not only affects the nutrient level but also the concentration of suspended organic and inorganic matter with enhanced water clarity and potentially better growth conditions for submerged macrophytes. We conducted a biomanipulation experiment in one of the basins in Chinese Huizhou West Lake that have remained highly turbid after extensive nutrient loading reduction. Another basin was used as control (control-treatment pairing design). Removal of a substantial amount of plankti-benthivorous fish was followed by planting of submerged macrophytes and stocking of piscivorous fish. We found strong and relatively long-lasting effects of the restoration initiative in the form of substantial improvements in water clarity and major reductions in nutrient concentrations, particularly total phosphorus, phytoplankton and turbidity, while only minor effects were detected for crustacean zooplankton grazers occurring in low densities before as well as after the restoration. Our results add importantly to the existing knowledge of restoration of warm lakes and are strongly relevant, not least in Asia where natural lakes frequently are used extensively for fish production, often involving massive stocking of benthivorous fish. With a growing economy and development of more efficient fish production systems, the interest in restoring lakes is increasing world-wide. We found convincing evidence that fish removal and piscivores stocking combined with transplantation of submerged macrophytes may have significant effects on water clarity in warm shallow lakes even if the zooplankton grazing potential remains low, the latter most likely as a result of high predation on the zooplankton.

Structural and functional responses of plant communities to climate change-mediated alterations in the hydrology of riparian areas in temperate Europe.

The hydrology of riparian areas changes rapidly these years because of climate change-mediated alterations in precipitation patterns. In this study, we used a large-scale in situ experimental approach to explore effects of drought and flooding on plant taxonomic diversity and functional trait composition in riparian areas in temperate Europe. We found significant effects of flooding and drought in all study areas, the effects being most pronounced under flooded conditions. In near-stream areas, taxonomic diversity initially declined in response to both drought and flooding (although not significantly so in all years) and remained stable under drought conditions, whereas the decline continued under flooded conditions. For most traits, we found clear indications that the functional diversity also declined under flooded conditions, particularly in near-stream areas, indicating that fewer strategies succeeded under flooded conditions. Consistent changes in community mean trait values were also identified, but fewer than expected. This can have several, not mutually exclusive, explanations. First, different adaptive strategies may coexist in a community. Second, intraspecific variability was not considered for any of the traits. For example, many species can elongate shoots and petioles that enable them to survive shallow, prolonged flooding but such abilities will not be captured when applying mean trait values. Third, we only followed the communities for 3 years. Flooding excludes species intolerant of the altered hydrology, whereas the establishment of new species relies on time-dependent processes, for instance the dispersal and establishment of species within the areas. We expect that altered precipitation patterns will have profound consequences for riparian vegetation in temperate Europe. Riparian areas will experience loss of taxonomic and functional diversity and, over time, possibly also alterations in community trait responses that may have cascading effects on ecosystem functioning.

Nitrogen and Phosphorus Removal from Agricultural Runoff in Integrated Buffer Zones.

Integrated buffer zones (IBZs) represent a novel form of edge-of-field technology in Northwest Europe. Contrary to the common riparian buffer strips, IBZs collect tile drainage water from agricultural fields by combining a ditch-like pond (POND), where soil particles can settle, and a flow-through filter bed (FILTERBED) planted with Alnus glutinosa (L.), a European alder (black alder). The first experimental IBZ facility was constructed and thoroughly tested in Denmark for its capability to retain various nitrogen (N) and phosphorus (P) species within the first three years after construction. We calculated the water and nutrient budget for the total IBZ and for the two compartments, POND and FILTERBED, separately. Furthermore, a tracer experiment using sodium bromide was conducted in order to trace the water flow and estimate the hydraulic residence time in the FILTERBEDs. The monthly average removal efficiency amounted to 10-67% for total N and 31-69% for total P, with performance being highest during the warm season. Accordingly, we suggest that IBZs may be a valuable modification of dry buffer strips in order to mitigate the adverse impacts of high nutrient loading from agricultural fields on the aquatic environment.

Headwater streams in the EU Water Framework Directive: Evidence-based decision support to select streams for river basin management plans.

Headwater streams are important contributors to aquatic biodiversity and may counteract negative impacts of anthropogenic stress on downstream reaches. In Denmark, the first river basin management plan (RBMP) included streams of all size categories, most being <2.5m wide (headwater streams). Currently, however, it is intensely debated whether the small size and low slopes, typical of Danish streams, in combination with degraded habitat conditions obstruct their ability to fulfill the ecological quality objectives required by the EU Water Framework Directive (WFD). The purpose of this study was to provide an analytically based framework for guiding the selection of headwater streams for RBMP. Specifically, the following hypotheses were addressed: i) stream slope, width, planform, and general physical habitat quality can act as criteria for selecting streams for the next generation of RBMPs, and ii) probability-based thresholds for reaching good ecological status can be established for some or all of these criteria, thus creating a sound, scientifically based, and clear selection process. The hypotheses were tested using monitoring data on Danish streams from the period 2004-2015. Significant linear relationships were obtained between the ecological quality ratio assessed by applying the Danish Stream Fauna Index (DSFIEQR) and stream slope, width, sinuosity, and DHI. The obtained models were used to produce pressure-response curves describing the probability of achieving good ecological status along gradients in these parameters. Next, threshold values for slope, width, sinuosity, and DHI were identified for selected probabilities of achieving minimum good ecological status. The obtained results can support managers and policy makers in prioritizing headwater streams for the 3rd RBMP. The approach applied is broadly applicable and can, for instance, help prioritization of restoration and conservation efforts in different types of ecosystems where the biota can be significantly linked to separate and quantifiable environmental characteristics.

Eutrophication effects on greenhouse gas fluxes from shallow-lake mesocosms override those of climate warming.

Fresh waters make a disproportionately large contribution to greenhouse gas (GHG) emissions, with shallow lakes being particular hot spots. Given their global prevalence, how GHG fluxes from shallow lakes are altered by climate change may have profound implications for the global carbon cycle. Empirical evidence for the temperature dependence of the processes controlling GHG production in natural systems is largely based on the correlation between seasonal temperature variation and seasonal change in GHG fluxes. However, ecosystem-level GHG fluxes could be influenced by factors, which while varying seasonally with temperature are actually either indirectly related (e.g. primary producer biomass) or largely unrelated to temperature, for instance nutrient loading. Here, we present results from the longest running shallow-lake mesocosm experiment which demonstrate that nutrient concentrations override temperature as a control of both the total and individual GHG flux. Furthermore, testing for temperature treatment effects at low and high nutrient levels separately showed only one, rather weak, positive effect of temperature (CH4 flux at high nutrients). In contrast, at low nutrients, the CO2 efflux was lower in the elevated temperature treatments, with no significant effect on CH4 or N2 O fluxes. Further analysis identified possible indirect effects of temperature treatment. For example, at low nutrient levels, increased macrophyte abundance was associated with significantly reduced fluxes of both CH4 and CO2 for both total annual flux and monthly observation data. As macrophyte abundance was positively related to temperature treatment, this suggests the possibility of indirect temperature effects, via macrophyte abundance, on CH4 and CO2 flux. These findings indicate that fluxes of GHGs from shallow lakes may be controlled more by factors indirectly related to temperature, in this case nutrient concentration and the abundance of primary producers. Thus, at ecosystem scale, response to climate change may not follow predictions based on the temperature dependence of metabolic processes.

Plant trait characteristics vary with size and eutrophication in European lowland streams.

Previous studies investigating community-level relationships between plant functional trait characteristics and stream environmental characteristics remain scarce. Here, we used community-weighted means to identify how plant traits link to lowland stream typology and how agricultural intensity in the catchment affects trait composition.We analysed plant trait characteristics in 772 European lowland streams to test the following two hypotheses: (i) trait characteristics differ between plant communities in small and medium-sized streams, reflecting adaptations to different habitat characteristics, and (ii) trait characteristics vary with the intensity of agricultural land use in the stream catchment, mediated either directly by an increase in productive species or indirectly by an increase in species that efficiently intercept and utilize light.We found that the communities in small streams were characterized by a higher abundance of light-demanding species growing from single apical meristems, reproducing by seeds and rooted to the bottom with floating and/or heterophyllous leaves, whereas the community in medium-sized streams was characterized by a higher abundance of productive species growing from multi-apical and basal growth meristems forming large canopies.We also found indications that community trait characteristics were affected by eutrophication. We did not find enhanced abundance of productive species with an increasing proportion of agriculture in the catchments. Instead, we found an increase in the abundance of species growing from apical and multi-apical growth meristems as well as in the abundance of species tolerant of low light availability. The increase in the abundance of species possessing these traits likely reflects different strategies to obtain greater efficiency in light interception and utilization in nutrient-enriched environments. Synthesis and applications. Our findings challenge the general assumption of the EU Water Framework Directive compliant assessment systems that plant community patterns in streams reflect the nutrient preference of the community. Instead, light availability and the ability to improve interception and utilization appeared to be of key importance for community composition in agricultural lowland streams. We therefore suggest moving from existing approaches building on species-specific preference values for nutrients to determine the level of nutrient impairment to trait-based approaches that provide insight into the biological mechanisms underlying the changes. We recommend that existing systems are critically appraised in the context of the findings of this study.

PXL01 in sodium hyaluronate for improvement of hand recovery after flexor tendon repair surgery: randomized controlled trial.

BACKGROUND: Postoperative adhesions constitute a substantial clinical problem in hand surgery. Fexor tendon injury and repair result in adhesion formation around the tendon, which restricts the gliding function of the tendon, leading to decreased digit mobility and impaired hand recovery. This study evaluated the efficacy and safety of the peptide PXL01 in preventing adhesions, and correspondingly improving hand function, in flexor tendon repair surgery. METHODS: This prospective, randomised, double-blind trial included 138 patients admitted for flexor tendon repair surgery. PXL01 in carrier sodium hyaluronate or placebo was administered around the repaired tendon. Efficacy was assessed by total active motion of the injured finger, tip-to-crease distance, sensory function, tenolysis rate and grip strength, and safety parameters were followed, for 12 months post-surgery. RESULTS: The most pronounced difference between the treatment groups was observed at 6 months post-surgery. At this timepoint, the total active motion of the distal finger joint was improved in the PXL01 group (60 vs. 41 degrees for PXL01 vs. placebo group, p = 0.016 in PPAS). The proportion of patients with excellent/good digit mobility was higher in the PXL01 group (61% vs. 38%, p = 0.0499 in PPAS). Consistently, the PXL01 group presented improved tip-to-crease distance (5.0 vs. 15.5 mm for PXL01 vs. placebo group, p = 0.048 in PPAS). Sensory evaluation showed that more patients in the PXL01 group felt the thinnest monofilaments (FAS: 74% vs. 35%, p = 0.021; PPAS: 76% vs. 35%, p = 0.016). At 12 months post-surgery, more patients in the placebo group were considered to benefit from tenolysis (30% vs. 12%, p = 0.086 in PPAS). The treatment was safe, well tolerated, and did not increase the rate of tendon rupture. CONCLUSIONS: Treatment with PXL01 in sodium hyaluronate improves hand recovery after flexor tendon repair surgery. Further clinical trials are warranted to determine the most efficient dose and health economic benefits. TRIAL REGISTRATION: ClinicalTrials.gov NCT01022242; EU Clinical Trials 2009-012703-25.

From expert judgement to supervised classification: a new approach to assess ecological status in lowland streams.

The EC Water Framework Directive (WFD) clearly states that undisturbed reference states of aquatic ecosystems should be used to set standards for restoration. Across Europe defining biological reference status and setting boundaries for ecological status classes continues to represent a major challenge. In the present study we investigate if a paradigm exists among experts that can guide the development of assessment systems based on the normative definitions of ecological status classes of the WFD. Our main questions were: 1) Will experts from species abundance data and typology descriptors independently arrive at similar assessments of ecological status, and 2) Can the expert interpretation of ecological status be transferred into a statistical model allowing for a standardization of assessments from plant assemblages in lowland streams? We used a large dataset covering 1244 randomly distributed stream sites in Denmark and asked a group of experts to independently classify the sites using the WFD's normative definitions of ecological status. According to the combined expert group, no Danish stream sites belonged to the undisturbed reference state. For the remaining ecological status classes we found good concordance in the classification made by the five experts. From this we infer that a common paradigm does exist, which may guide the development of assessment methods for aquatic plants in lowland streams. We also found that the common view of the experts could be transferred into a supervised classification model that can serve as a classification tool for aquatic plant assemblages in lowland streams. We conclude that the combined use of experts and advanced multivariate statistics can provide a useful approach in the development of systems for assessment of ecological status in water types, where a reference network cannot be established.

Sampling of herbicides in streams during flood events.

In stream water xenobiotics usually occur as pulses in connection with floods caused by surface run-off and tile drainage following precipitation events. In streams located in small agricultural catchments we monitored herbicide concentrations during flood events by applying an intensive sampling programme of ½ h intervals for 7 h. In contrast to grab sampling under non-flood conditions, clearly elevated concentrations were recorded during the floods, and pulses varying in occurrence, duration and concentration were recorded. Pulses of recently applied herbicides were the most prominent, but also agricultural herbicides used in previous seasons caused pulses in the streams. Asynchronism of chemographs may be related to the characteristics of the compounds as well as their transport pathways and transformation in compartments between the source and the point of sampling in the stream. Thus, the occurrence of chemographs is difficult to predict, which ought to be taken into account when designing a sampling strategy. Even though the chemographs of herbicides and their transformation products (glyphosate and aminomethylphosphonic acid (AMPA) as well as terbuthylazine and desethylterbuthylazine) seem to be synchronous, their occurrence may still be difficult to predict. It is evident that grab sampling under non-flood conditions yields insufficient information on the dynamics of occurrence of herbicides in stream water, both with respect to environmental effects and the calculation of the load to a recipient. In conclusion, the design of a sampling strategy regarding herbicides in stream waters should adequately consider the aim of the investigation.

Phosphorus load to surface water from bank erosion in a Danish lowland river basin.

Phosphorus loss from bank erosion was studied in the catchment of River Odense, a lowland Danish river basin, with the aim of testing the hypothesis of whether stream banks act as major diffuse phosphorus (P) sources at catchment scale. Furthermore, the study aimed at analyzing the impact of different factors influencing bank erosion and P loss such as stream order, anthropogenic disturbances, width of uncultivated buffer strips, and the vegetation of buffer strips. A random stratified procedure in geographical information system (GIS) was used to select two replicate stream reaches covering different stream orders, channelized vs. naturally meandering channels, width of uncultivated buffer strips (≤ 2 m and ≥ 10 m), and buffer strips with different vegetation types. Thirty-six 100-m stream reaches with 180 bank plots and a total of 3000 erosion pins were established in autumn 2006, and readings were conducted during a 3-yr period (2006-2009). The results show that neither stream size nor stream disturbance measured as channelization of channel or the width of uncultivated buffer strip had any significant ( < 0.05) influence on bank erosion and P losses during each of the 3 yr studied. In buffer strips with natural trees bank erosion was significantly ( < 0.05) lower than in buffer strips dominated by grass and herbs. Gross and net P input from bank erosion amounted to 13.8 to 16.5 and 2.4 to 6.3 t P, respectively, in the River Odense catchment during the three study years. The net P input from bank erosion equaled 17 to 29% of the annual total P export and 21 to 62% of the annual export of P from diffuse sources from the River Odense catchment. Most of the exported total P was found to be bioavailable (71.7%) based on a P speciation of monthly suspended sediment samples collected at the outlet of the river basin. The results found in this study have a great importance for managers working with P mitigation and modeling at catchment scale.

A distributed modelling system for simulation of monthly runoff and nitrogen sources, loads and sinks for ungauged catchments in Denmark.

We validated an existing physically based 3D MIKE SHE groundwater resource model (DK-model) at 175 Danish gauging stations covering different catchment sizes in order to calculate monthly water runoff in the 50% ungauged part of Denmark. Model performance was in most cases good (61% of gauging stations had a Nash-Sutcliffe (NS) coefficient >0.60) but nevertheless showed a large seasonal and georegion specific bias. Therefore, bias correction factors had to be developed before applying the DK-model simulations of runoff in the ungauged areas. Simulated monthly runoff from ungauged areas and the measured monthly runoff from 178 gauging stations were distributed to 2663 smaller Hydrological Units (ca. 15 km(2)) and linked with a new empirical model for flow-weighted monthly total nitrogen (TN) concentrations (R(2) = 0.43; P < 0.0001) developed based on 20 years of observations (1990-2009) in 83 small catchments for calculation of monthly gross diffuse TN-loads from HU's. Nitrogen retention was calculated in streams, lakes and wetlands utilising both lake specific models and rate coefficients to calculate N retention in surface water bodies. The whole model complex was linked in the DK-QN concept for simulation of monthly TN losses from point sources and diffuse sources, TN retention and resulting loadings to Danish coastal waters. The DK-QN model was validated in 118 gauged catchments and the model simulations had for >25% of the observations of monthly discharge weighted TN concentrations a NS larger than 0.26. Catchment specific monthly TN-loadings were modelled with a higher performance as 50% of the catchments had a NS greater than 0.75. The model concept allows calculation of N retention in streams, lakes and wetlands and the average annual model calculated N retention amounted to 21% of the modelled gross riverine TN loadings. The average annual gross TN loading to surface freshwater in Denmark derived from diffuse sources amounted to 97 000 tonnes N (91% of gross TN loadings) which is 54% of the total estimated N-leaching from the root zone on the Danish land area (212 000 tonnes N) during the period 1990-2009.

Impact of lambda-cyhalothrin on a macroinvertebrate assemblage in outdoor experimental channels: implications for ecosystem functioning.

In this study, the impact of a single pulse of the pyrethroid lambda-cyhalothrin was tested on a macroinvertebrate assemblage consisting of Gammarus pulex, Leuctra nigra, Heptagenia sulphurea and Ancylus fluviatilis in outdoor experimental stream channels. Channels (4m long, 0.1m wide) were groundwater fed and had natural substratum. Macroinvertebrates were exposed to 10.65 or 106.5 ng L(-1) lambda cyhalothrin for 90 min in the laboratory and after 24h introduced to the experimental stream channels with four replicates of each treatment and controls. Drift samples were taken with 24-h interval for 10 days and behaviour of drifted macroinvertebrates was assessed. Microalgae biomass was measured on days 1, 5, 8 and 10 along with leaf litter decomposition using leaf packs of beech (Fagus sylvatica). Numbers of drifting G. pulex and L. nigra with reduced mobility increased significantly with concentration of lambda-cyhalothrin. Increase of algal biomass was significantly greater in stream channels with macroinvertebrates exposed to 106.5 ng L(-1) compared to controls and 10.65 ng L(-1) treatments. Accrual of microalgal biomass was significantly higher in the high concentration treatment and decomposition of leaf litter was significantly greater in control channels compared to channels with exposed macroinvertebrates. This study may apply valuable knowledge to the understanding and assessment of how pyrethroids impact ecosystem functioning in streams.

The search for reference conditions for stream vegetation in northern Europe.

1. The European Water Framework Directive provides a framework for improving the ecological quality of stream ecosystems, with deviation from reference used as a measure of ecological status. 2. Here we examine the possibility of using less impacted stream sites from Latvia, Lithuania and Poland to establish a Danish reference network for macrophyte assemblages, and as a guiding image for identification of possible references sites within Denmark. Both approaches were evaluated using historical Danish records. 3. Four different macrophyte assemblages were identified for mid-sized streams in the Central and Eastern Lowland ecoregions. Macrophyte assemblages could not be delineated using physical stream site characteristics; however a gradual change in assemblage composition was attributed to differences in alkalinity and human impact. 4. Assemblages of contemporary vegetation in Denmark were quite similar to those found in Polish, Latvian and Lithuanian streams (26-35%). However, more importantly, from species-based predictions we noted higher similarity, particularly with Latvian and Lithuanian streams, before intensive land use commenced in Denmark (c. 1900). These results show that stream sites from these three countries can be considered in a Danish reference network. 5. Two of the four macrophyte assemblages comprised species such as Fontinalis antipyretica, Myriophyllum spicatum, Nuphar lutea, Potamogeton alpinus and P. perfoliatus that have a very scattered occurrence in the contemporary vegetation in Denmark. These groups were closely associated with the predictions from historic records, thereby lending support the conjecture that these assemblages could be part of the guiding image for the identification of potential reference sites within Denmark.

Climate-change impacts on hydrology and nutrients in a Danish lowland river basin.

The Mike 11-TRANS modelling system was applied to the lowland Gjern river basin in Denmark to assess climate-change impacts on hydrology and nitrogen retention processes in watercourses, lakes and riparian wetlands. Nutrient losses from land to surface waters were assessed using statistical models incorporating the effect of changed hydrology. Climate-change was predicted by the ECHAM4/OPYC General Circulation Model (IPCC A2 scenario) dynamically downscaled by the Danish HIRHAM regional climate model (25 km grid) for two time slices: 1961-1990 (control) and 2071-2100 (scenario). HIRHAM predicts an increase in mean annual precipitation of 47 mm (5%) and an increase in mean annual air temperature of 3.2 degrees C (43%). The HIRHAM predictions were used as external forcings to the rainfall-runoff model NAM, which was set up and run for 6 subcatchments within and for the entire, Gjern river basin. Mean annual runoff from the river basin increases 27 mm (7.5%, p<0.05) when comparing the scenario to the control. Larger changes, however, were found regarding the extremes; runoff during the wettest year in the 30-year period increased by 58 mm (12.3%). The seasonal pattern is expected to change with significantly higher runoff during winter. Summer runoff is expected to increase in predominantly groundwater fed streams and decrease in streams with a low base-flow index. The modelled change in the seasonal hydrological pattern is most pronounced in first- or second-order streams draining loamy catchments, which currently have a low base-flow during the summer period. Reductions of 40-70% in summer runoff are predicted for this stream type. A statistical nutrient loss model was developed for simulating the impact of changed hydrology on diffuse nutrient losses (i.e. losses from land to surface waters) and applied to the river basin. The simulated mean annual changes in TN loads in a loamy and a sandy subcatchment were, respectively, +2.3 kg N ha(-1) (8.5%) and +1.6 kg N ha(-1) (6.9%). The rainfall-runoff model and the nutrient loss model were chained with Mike 11-TRANS to simulate the combined effects of climate-change on hydrology, nutrient losses and nitrogen retention processes at the scale of the river basin. The mean annual TN export from the river basin increased from the control to the scenario period by 7.7%. Even though an increase in nitrogen retention in the river system of 4.2% was simulated in the scenario period, an increased in-stream TN export resulted because of the simulated increase in the diffuse TN transfer from the land to the surface-waters.