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Anthropogenically forced changes in global freshwater biodiversity demand more efficient monitoring approaches. Consequently, environmental DNA (eDNA) analysis is enabling ecosystem-scale biodiversity assessment, yet the appropriate spatio-temporal resolution of robust biodiversity assessment remains ambiguous. Here, using intensive, spatio-temporal eDNA sampling across space (five rivers in Europe and North America, with an upper range of 20–35 km between samples), time (19 timepoints between 2017 and 2018) and environmental conditions (river flow, pH, conductivity, temperature and rainfall), we characterise the resolution at which information on diversity across the animal kingdom can be gathered from rivers using eDNA. In space, beta diversity was mainly dictated by turnover, on a scale of tens of kilometres, highlighting that diversity measures are not confounded by eDNA from upstream. Fish communities showed nested assemblages along some rivers, coinciding with habitat use. Across time, seasonal life history events, including salmon and eel migration, were detected. Finally, effects of environmental conditions were taxon-specific, reflecting habitat filtering of communities rather than effects on DNA molecules. We conclude that riverine eDNA metabarcoding can measure biodiversity at spatio-temporal scales relevant to species and community ecology, demonstrating its utility in delivering insights into river community ecology during a time of environmental change. Using intensive eDNA sampling in space and time across five rivers in Europe and North America, this study shows that eDNA gives relevant information on freshwater diversity and ecology across broad taxonomic groups, and with limited downstream transport. The findings demonstrate that eDNA is vital for freshwater biodiversity monitoring in a time of anthropogenic change.

Rapidly assessing biodiversity is essential for environmental monitoring; however, traditional approaches are limited in the scope needed for most ecological systems. Environmental DNA (eDNA) based assessment offers enhanced scope for assessing biodiversity, while also increasing sampling efficiency and reducing processing time, compared to traditional methods. Here we investigated the effects of landuse and seasonality on headwater community richness and functional diversity, via spatio-temporal dynamics, using both eDNA and traditional sampling. We found that eDNA provided greater resolution in assessing biodiversity dynamics in time and space, compared to traditional sampling. Community richness was seasonally linked, peaking in spring and summer, with temporal turnover having a greater effect on community composition compared to localized nestedness. Overall, our assessment of ecosystem function shows that community formation is driven by regional resource availability, implying regional management requirements should be considered. Our findings show that eDNA based ecological assessment is a powerful, rapid and effective assessment strategy that enables complex spatio-temporal studies of community diversity and ecosystem function, previously infeasible using traditional methods.Mathew Seymour et al. compare eDNA with traditional biodiversity metrics to assess the functional diversity of a river basin in Wales over space and time and variable land use. Their results show that eDNA can generate greater biodiversity resolution and reliably detect spatio-temporal changes in community and functional diversity.

Groundwater ecosystems remain poorly understood yet may provide ecosystem services, make a unique contribution to biodiversity and contain useful bio-indicators of water quality. Little is known about ecosystem variability, the distribution of invertebrates within aquifers, or how representative boreholes are of aquifers. We addressed these issues using borehole imaging and single borehole dilution tests to identify three potential aquifer habitats (fractures, fissures or conduits) intercepted by two Chalk boreholes at different depths beneath the surface (34 to 98 m). These habitats were characterised by sampling the invertebrates, microbiology and hydrochemistry using a packer system to isolate them. Samples were taken with progressively increasing pumped volume to assess differences between borehole and aquifer communities. The study provides a new conceptual framework to infer the origin of water, invertebrates and microbes sampled from boreholes. It demonstrates that pumping 5 m(3) at 0.4-1.8 l/sec was sufficient to entrain invertebrates from five to tens of metres into the aquifer during these packer tests. Invertebrates and bacteria were more abundant in the boreholes than in the aquifer, with associated water chemistry variations indicating that boreholes act as sites of enhanced biogeochemical cycling. There was some variability in invertebrate abundance and bacterial community structure between habitats, indicating ecological heterogeneity within the aquifer. However, invertebrates were captured in all aquifer samples, and bacterial abundance, major ion chemistry and dissolved oxygen remained similar. Therefore the study demonstrates that in the Chalk, ecosystems comprising bacteria and invertebrates extend from around the water table to 70 m below it. Hydrogeological techniques provide excellent scope for tackling outstanding questions in groundwater ecology, provided an appropriate conceptual hydrogeological understanding is applied.

Climatic changes could transform rivers as drought becomes more frequent with potentially severe, but largely unknown, consequences at multispecies levels of organization. Now research shows experimentally how the intensification of drought may alter the underlying structure and functioning of freshwater food webs. Climate change is expected to make many regions of the world much drier over coming decades 1 , 2 . More intense drought would transform rivers 3 with potentially severe but largely unknown consequences at higher (multispecies) levels of organization 4 . Here we show experimentally how the intensification of drought may alter the underlying structure and functioning (biomass flux dynamics) of freshwater food webs—networks of species and their interactions 5 . Drought triggered substantial losses of species and links, especially among predators, leading to the partial collapse of the food webs. Total resource–consumer biomass flux was also strongly suppressed by disturbance, yet several network-level properties (such as connectance and interaction diversity) were conserved, driven by consumer resource fidelity and a substantial reconfiguration of fluxes within the webs as production shifted down the size spectrum from large to small species. Our research demonstrates that drier climates could have far-reaching impacts on the functioning of freshwater ecosystems.

Accurate quantification of biodiversity is fundamental to understanding ecosystem function and for environmental assessment. Molecular methods using environmental DNA (eDNA) offer a non-invasive, rapid, and cost-effective alternative to traditional biodiversity assessments, which require high levels of expertise. While eDNA analyses are increasingly being utilized, there remains considerable uncertainty regarding the dynamics of multispecies eDNA, especially in variable systems such as rivers. Here, we utilize four sets of upland stream mesocosms, across an acid–base gradient, to assess the temporal and environmental degradation of multispecies eDNA. Sampling included water column and biofilm sampling over time with eDNA quantified using qPCR. Our findings show that the persistence of lotic multispecies eDNA, sampled from water and biofilm, decays to non-detectable levels within 2 days and that acidic environments accelerate the degradation process. Collectively, the results provide the basis for a predictive framework for the relationship between lotic eDNA degradation dynamics in spatio-temporally dynamic river ecosystems. Mathew Seymour et al. investigate the persistence of environmental DNA (eDNA) in river systems in environments of varying pH. Using four sets of upland stream mesocosms, they find that eDNA degrades to non-detectable levels within two days and this degradation is accelerated in acidic environments.

High-resolution monitoring of water quality and ecosystem functioning over large spatial scales in expansive lowland river catchments is challenging. Therefore, we need modeling tools to predict these processes at locations where observations are absent. Here, we present a new approach to estimate ecosystem metabolism underpinned by a high-resolution, process-based model of in-stream flows and water quality. The model overcomes the current challenges in metabolism modeling by accounting for oxygen transport under varying flows and oxygen transformations due to biogeochemical processes. We implement the model in a 62-km-long stretch of the River Thames, England, using observations spanning 2 yr. Model outputs suggest that the river is primarily autotrophic from mid-spring to mid-summer due to high biomass during low-flow periods, and is heterotrophic during the rest of the year. Ecosystem respiration in upstream reaches is driven mainly by biochemical oxygen demand, autotrophic respiration, and nitrification processes, whereas downstream sites also show a control of benthic oxygen demand in addition to the aforementioned processes. Using empirical modeling, we analyze the sensitivity of our estimated metabolism rates to multiple environmental stressors. Results demonstrate that empirical models could be useful for rapid river health assessments, but need improvements to reproduce peak metabolism rates. The process-based model, although more complex than existing in situ approaches to metabolism quantification, allows inference when gaps in continuous observations are present. The model offers additional benefits for predicting metabolism rates under future scenarios of environmental change incorporating multiple stressor effects.

The subphylum Crustacea includes the most successful species among aquatic alien invaders. The impacts of invasive alien crustaceans (IAC) are often substantial, due to the complex trophic role of most of these species leading to cascading effects throughout the invaded ecosystems. IAC also have the potential to cause a shift in the ‘keystone’ ecosystem functions, changing energy flux and nutrient cycles which together affect critical ecosystem services such as biodiversity, fisheries yield and water quality. Although no individual trait appears to be a good predictor of invasion success, a combination of some characteristics such as eurytolerance, omnivory and certain r-selected life-history traits results in a high probability of alien crustacean species becoming invasive. Both environmental factors, such as habitat heterogeneity in the invaded ecosystems, and evolutionary factors, such as adaptations to new environmental conditions, also play important roles during establishment. Therefore, individual environmental niche models, including genetic algorithm, have the highest likelihood of providing useful predictive information about invasion success and spread of alien Crustacea. Attempts to control IAC through biocides or mechanical removal have had mixed success in the past but a strategic combination of different methods may lead to some success in the future.

The Glastir Monitoring and Evaluation Programme (GMEP) ran from 2013 until 2016 and was probably the most comprehensive programme of ecological study ever undertaken at a national scale in Wales. The programme aimed to (1) set up an evaluation of the environmental effects of the Glastir agri-environment scheme and (2) quantify environmental status and trends across the wider countryside of Wales. The focus was on outcomes for climate change mitigation, biodiversity, soil and water quality, woodland expansion, and cultural landscapes. As such, GMEP included a large field-survey component, collecting data on a range of elements including vegetation, land cover and use, soils, freshwaters, birds, and insect pollinators from up to three-hundred 1 km survey squares throughout Wales. The field survey capitalised upon the UK Centre for Ecology & Hydrology (UKCEH) Countryside Survey of Great Britain, which has provided an extensive set of repeated, standardised ecological measurements since 1978. The design of both GMEP and the UKCEH Countryside Survey involved stratified-random sampling of squares from a 1 km grid, ensuring proportional representation from land classes with distinct climate, geology and physical geography. Data were collected from different land cover types and landscape features by trained professional surveyors, following standardised and published protocols. Thus, GMEP was designed so that surveys could be repeated at regular intervals to monitor the Welsh environment, including the impacts of agri-environment interventions. One such repeat survey is scheduled for 2021 under the Environment and Rural Affairs Monitoring & Modelling Programme (ERAMMP). Data from GMEP have been used to address many applied policy questions, but there is major potential for further analyses. The precise locations of data collection are not publicly available, largely for reasons of landowner confidentiality. However, the wide variety of available datasets can be (1) analysed at coarse spatial resolutions and (2) linked to each other based on square-level and plot-level identifiers, allowing exploration of relationships, trade-offs and synergies. This paper describes the key sets of raw data arising from the field survey at co-located sites (2013 to 2016). Data from each of these survey elements are available with the following digital object identifiers (DOIs): Landscape features (Maskell et al., 2020a–c), https://doi.org/10.5285/82c63533-529e-47b9-8e78-51b27028cc7f, https://doi.org/10.5285/9f8d9cc6-b552-4c8b-af09-e92743cdd3de, https://doi.org/10.5285/f481c6bf-5774-4df8-8776-c4d7bf059d40; Vegetation plots (Smart et al., 2020), https://doi.org/10.5285/71d3619c-4439-4c9e-84dc-3ca873d7f5cc; Topsoil physico-chemical properties (Robinson et al., 2019), https://doi.org/10.5285/0fa51dc6-1537-4ad6-9d06-e476c137ed09; Topsoil meso-fauna (Keith et al., 2019), https://doi.org/10.5285/1c5cf317-2f03-4fef-b060-9eccbb4d9c21; Topsoil particle size distribution (Lebron et al., 2020), https://doi.org/10.5285/d6c3cc3c-a7b7-48b2-9e61-d07454639656; Headwater stream quality metrics (Scarlett et al., 2020a), https://doi.org/10.5285/e305fa80-3d38-4576-beef-f6546fad5d45; Pond quality metrics (Scarlett et al., 2020b), https://doi.org/10.5285/687b38d3-2278-41a0-9317-2c7595d6b882; Insect pollinator and flower data (Botham et al., 2020), https://doi.org/10.5285/3c8f4e46-bf6c-4ea1-9340-571fede26ee8; and Bird counts (Siriwardena et al., 2020), https://doi.org/10.5285/31da0a94-62be-47b3-b76e-4bdef3037360.

1. A fundamental goal of ecological network research is to understand how the complexity observed in nature can persist and how this affects ecosystem functioning. This is essential for us to be able to predict, and eventually mitigate, the consequences of increasing environmental perturbations such as habitat loss, climate change, and invasions of exotic species. 2. Ecological networks can be subdivided into three broad types: 'traditional' food webs, mutualistic networks and host-parasitoid networks. There is a recent trend towards cross-comparisons among network types and also to take a more mechanistic, as opposed to phenomenological, perspective. For example, analysis of network configurations, such as compartments, allows us to explore the role of co-evolution in structuring mutualistic networks and host-parasitoid networks, and of body size in food webs. 3. Research into ecological networks has recently undergone a renaissance, leading to the production of a new catalogue of evermore complete, taxonomically resolved, and quantitative data. Novel topological patterns have been unearthed and it is increasingly evident that it is the distribution of interaction strengths and the configuration of complexity, rather than just its magnitude, that governs network stability and structure. 4. Another significant advance is the growing recognition of the importance of individual traits and behaviour: interactions, after all, occur between individuals. The new generation of high-quality networks is now enabling us to move away from describing networks based on species-averaged data and to start exploring patterns based on individuals. Such refinements will enable us to address more general ecological questions relating to foraging theory and the recent metabolic theory of ecology. 5. We conclude by suggesting a number of 'dead ends' and 'fruitful avenues' for future research into ecological networks.

Chalk streams are among the most species-rich and productive of all temperate ecosystems. Despite this, a few keystone species have the potential to exert disproportionately powerful effects on community structure and ecosystem processes. Two of these are the bullhead Cottus gobio, a small benthic fish that is an extremely abundant, voracious predator, and the freshwater shrimp Gammaruspulex, which dominates the prey assemblage and is the principal detritivore. Field experiments detected a bullhead-Gammarus-detritus trophic cascade, with detrital processing rates slowed dramatically in the presence of the predator. In addition, survey data also revealed strong negative density-dependence between bullhead and brown trout, adding a further link in the cascade. However, although bullhead also depressed the abundance of a dominant grazer, the snail Potamopyrgus antipodarum, there was no cascading effect upon algal production, suggesting that autochthonous inputs were not controlled by top-down effects. This skewed effect of the predator upon autochthonous versus allochthonous basal resources stresses the need to consider both pathways of energy flux into the food web, whereas many previous studies have potentially overemphasized the importance of predator-herbivore-primary producer cascades. The wider community food web contained 142 species and 1383 feeding links. This complex network exhibited \"small world\" properties, such as high clustering (unlike many other food webs) and shortest path lengths between species were small (in common with many other food webs). In particular, each of the four members of the detrital cascade could be connected to any other species by three links or fewer. Our data revealed that powerful cascading effects can be imbedded within even very complex ecological networks.