The invasion of the North Atlantic by pink salmon Oncorhynchus gorbuscha has raised concerns regarding their impact on coastal rivers. Although the influence of marine-derived nutrients from returning adult O. gorbuscha on rivers in their native range has received much attention, the ecological consequences of invasive O. gorbuscha for ecosystems outside the native range are largely unknown. To investigate the impact on the density and community structure of benthic macroinvertebrates, O. gorbuscha carcasses were added to 12 experimental channels for 60 days at three treatment levels (control, no carcass; low and high, loading rates). Stable isotopes of carbon (δ13C) and nitrogen (δ15N) were used to determine if nutrients from carcasses were incorporated into native biota. The density of macroinvertebrates increased close to the carcasses in the high-addition treatment, suggesting aggregation. Furthermore, macroinvertebrates had a higher δ15N near to the carcasses in the low- and high-addition treatments after 30 days, indicating uptake from the carcasses. The higher δ15N of willow moss Fontinalis antipyretica in carcass-addition treatments indicated that primary producers could also assimilate nutrients from the decomposition of carcasses. Whilst the addition of carcasses resulted in the increased density of small individuals of macroinvertebrates, this did not propagate to changes in community composition in this relatively short experiment. Overall, the results suggest that native biota might benefit from the marine-derived nutrients transported to streams by invasive O. gorbuscha, however, the long-term effects of such nutrient/energy subsidies on receiving ecosystems require further investigation.

Advancements have been made in the use of DNA-based methods for the detection of single species. However, the routine application of DNA-based methods to monitor whole communities using a metabarcoding approach and derive ecosystem status continues to be limited. We undertook a structured experiment to assess factors influencing the precision and accuracy of molecular methods for freshwater macroinvertebrate community assessment. Macroinvertebrates were sorted, identified and counted from kick-net samples using standard morphometric protocols, then reconstituted. These bulk specimen samples were then homogenised. From each bulk sample, aliquots of the homogenate were distributed among seven laboratories across Europe for DNA extraction, PCR amplification, library preparation and sequencing. Additionally, each laboratory was provided with a DNA extract from the bulk homogenate to allow for amplification through to sequencing, in order to assess the influence of DNA extraction. Hierarchical nested analyses were used to identify sources of uncertainty in the data and explore factors influencing the returned community and metrics of ecosystem quality. Furthermore, comparison with morphological analysis enabled us to benchmark molecular to traditional processing approaches. Metrics derived by morphological and DNA-based methods differed substantially. The strongest methodological influences on variance in metrics derived from DNA-based methods were attributed to sequencing (read count per sample) and laboratory (strong variation within samples). Furthermore, the performance of the DNA-based method varied among samples, most likely due to differences in the composition of the invertebrate community. Logistic regression indicated that detectability did not vary among taxa, but that taxa occurring in low abundance (less than five individuals in our samples) were more likely to be missed via metabarcoding. Our findings highlight sources of uncertainty influencing DNA-based methods and, particularly, the variability of results generated from individual laboratories. Our findings underline the need for clear guidelines, standardisation and quality assurance schemes.

Monitoring programmes worldwide use biota to assess the "health" of water bodies. Indices based on biota are used to describe the change in status of sites over time, to identify progress against management targets and to diagnose the causes of biological degradation. A variety of numerical stressor-specific biotic indices have been developed based on the response of biota to differences in stressors among sites. Yet, it is not clear how variation in pressures within sites, over what time period, and in what combination has the greatest impact on different biotic groups. An understanding of how temporal variation in pressures influences biological assessment indices would assist in setting achievable targets and help focus catchment-scale mitigation strategies to ensure that they deliver the desired improvements in biological condition. Hydrochemical data provided by a network of high-frequency (15 or 30 min) automated monitoring stations over 3 years were matched to replicated biological data to understand the influence of spatio-temporal variation in pollution pressures on biological indices. Hydrochemical data were summarised in various ways to reflect central tendency, peaks, troughs and variation over 1-90 days before the collection of each biological sample. An objective model selection procedure was used to determine which hydrochemical determinand, and over what time period, best explained variation in the biological indices. Stressor-specific indices derived from macroinvertebrates which purportedly assess stress from low flows, excess fine sediment, nutrient enrichment, pesticides and organic pollution were significantly inter-correlated and reflected periods of low oxygen concentration, even though only one index (ASPT(WHPT), average score per taxon) was designed for this purpose. Changes in community composition resulting from one stressor frequently lead to confounding effects on stressor-specific indices. Variation in ASPT(WHPT) was best described by dissolved oxygen calculated as Q(5) over 10 days, suggesting that low oxygen events had most influence over this period. Longer-term effects were apparent, but were masked by recovery. Macroinvertebrate abundance was best described by Q(95) of stream velocity over 60 days, suggesting a slower recovery in numbers than in the community trait reflected by ASPT(WHPT). Although use of ASPT(WHPT) was supported, we recommend that additional independent evidence should be used to corroborate any conclusions regarding the causes of degradation drawn from the other stressor-specific indices. The use of such stressor-specific indices alone risks the mistargeting of management strategies if the putative stressor-index approach is taken to be more reliable than the results herein suggest.

Historical mining activities cause widespread, long-term trace metal contamination of freshwater ecosystems. However, measuring trace metal bioavailability has proven difficult, because it depends on many factors, not least concentrations in water, sediment and habitat. Simple tools are needed to assess bioavailabilities. The use of biomonitors has been widely advocated to provide a realistic measure. To date there have been few attempts to identify ubiquitous patterns of trace metal accumulation within and between freshwater biomonitors at geographical scales relevant to trace metal contamination. Here we address this through a nationwide collection of freshwater biomonitors (species of Gammarus, Leuctra, Baetis, Rhyacophila, Hydropsyche) from 99 English and Welsh stream sites spanning a gradient of high to low trace metal loading. The study tested for inter-biomonitor variation in trace metal body burden, and for congruence amongst accumulations of trace metals within taxa and between taxa across the gradient. In general, significant differences in trace metal body burden occurred between taxa: Gammarus sp. was the most different compared with insect biomonitors. Bivariate relationships between trace metals within biomonitors reflected trace metal profiles in the environment. Strong correlations between some trace metals suggested accumulation was also influenced by physiological pathways. Bivariate relationships between insect biomonitors for body burdens of As, Cu, Mn and Pb were highly consistent. Our data show that irrespective of taxonomic or ecological differences, there is a commonality of response amongst insect taxa, indicating one or more could provide consistent measures of trace metal bioavailability.

We used the systematic review procedure to assess the evidence available on the analysis, prevalence and impact of microplastics in freshwater and estuarine environments. As the study of microplastics in freshwaters is relatively new, measurement methods are yet to be standardized, and a wide variety of methods of variable robustness have been used. Critically, the sampling methodology used in the literature had a systematic influence on the concentration of microplastic particles returned. The volume of water sampled varied over many orders of magnitude, and there was a direct relationship between the size of the smallest particles studied and the volume of water sampled in both freshwater and estuaries: large volumes of water can only be sampled using nets of relatively coarse mesh, which in turn do not capture smaller particles. Consequently, the mean abundance of microplastic particles reported was inversely correlated with both the volume of water sampled and the size of particles studied. The size of microplastic particles had a substantial and overriding effect on threshold concentrations above which microplastics affect freshwater and estuarine biota. For the ecotoxicological endpoints of feeding, behaviour, growth and survival there was a clear relationship between the size of the particles used in the test and the threshold concentration at which an effect was seen. Although the taxonomic coverage of test organisms was limited, there were sufficient data to test the influence of taxonomic group used on size-specific thresholds for Crustacea, fish and algae. There was no significant effect of either the endpoint measured or the taxonomic group used, suggesting that there might not be any difference in sensitivity among different taxa. In order to establish a threshold concentration where microplastics present a hazard to a limited number of taxa, quantile regression was used to determine the size-specific concentration of microplastics that was lower than 90% of the thresholds identified for survival and, as a more conservative limit, across all endpoints tested including sublethal effects. By comparing these thresholds with the data on concentrations of microplastics reported by field studies, it was apparent that the calculated size specific threshold concentration for lethal effects was considerably higher than 99% of reported environmental concentrations. Lethal effects of microplastics on freshwater and estuarine biota are likely to be limited to exceptional circumstances. Over certain size ranges the calculated size specific threshold concentration for sublethal effects was exceeded by the highest 10% of concentrations reported from environmental samples, suggesting that there is a risk of sublethal effects in a small proportion of sites.

Despite growing interest in the environmental impact of microplastics, a standardized characterization method is not available. We carried out a systematic analysis of reliable global data detailing the relative abundance of polymers in freshwaters and estuaries. The polymers were identified according to seven main categories: polyethylene terephthalate, polyethylene, polyvinyl chloride, polypropylene, polystyrene, polyurethane and a final category of miscellaneous plastic. The results show that microplastics comprised of polyvinyl chloride and polyurethane are significantly less abundant than would be expected based on global production, possibly due to their use. This has implications for models of microplastic release into the environment based on production and fate. When analysed by matrix (water, sediment or biota) distinct profiles were obtained for each category. Polyethylene, polypropylene and polystyrene were more abundant in sediment than in biota, while miscellaneous plastics was more frequent in biota. The data suggest that environmental sorting of microplastic particles, influenced by physical, chemical and biological processes, may play a key role in environmental impact, although partitioning among matrices based on density was not realized. The distinct profile of microplastics in biota raises an important question regarding potential selectivity in uptake by organisms, highlighting the priority for more and better-informed laboratory exposure studies.

Metal-rich sediments have the potential to impair life in freshwater streams and rivers and, thereby, to inhibit recovery of ecological conditions after any remediation of mine water discharges. Sediments remain metal-rich over long time periods and have long-term potential ecotoxicological interactions with local biota, unless the sediments themselves are physically removed or replaced by less metal-rich sediment. Laboratory-derived environmental quality standards are difficult to apply to the field situation, as many complicating factors exist in the real world. Therefore, there is a strong case to consider other, field-relevant, measures of toxic effects as alternatives to laboratory-derived standards and to seek better biological tools to detect, diagnose and ideally predict community-level ecotoxicological impairment. Hence, this review concentrated on field measures of toxic effects of metal-rich sediment in freshwater streams, with less emphasis on laboratory-based toxicity testing approaches. To this end, this review provides an overview of the impact of metal-rich sediments on freshwater stream life, focusing on biological impacts linked to metal contamination.

Bovington Camp has been a main training area for armoured vehicles since 1916. Over the years, this has resulted in the erosion and mobilisation of soil particles into watercourses draining the area. Sediment-rich water from a main tributary, the Bovington Stream, discolours the River Frome during high flows. Regulatory agencies and angling groups were concerned that the quality of the river was being affected by run-off from the military ranges. Macroinvertebrate communities were studied between 1998 and 2016 at two sites on the Bovington Stream above and below a sediment trapping pond and two sites on the River Frome above and below the confluence. Samples were collected in Spring and Autumn and ecological quality was assessed using the River Invertebrate Classification Tool (RICT). There were marked increases in the ecological quality of the Bovington sites over time with most change occurring in the period 1998 to 2004. The two Frome sites were consistently classed as high quality throughout the study period. Increases in taxon richness between 1998–2004 and 2011–2016 were significant in the Bovington Stream but no change was observed in the River Frome. Faunal abundance at all sites fluctuated but there were no significant trends. The Combined Fine Sediment Index (CoFSI), assesses the impact of fine sediment pressure on the macroinvertebrate community. Low CoFSI scores were recorded in the early years of the study (1998–2000) at both Bovington sites but showed a steady increase until around 2010–2011, suggesting a degree of improvement/recovery from fine sediment pressure. No similar changes in CoFSI scores occurred in the River Frome. In the absence of any major disturbances in the Bovington catchment other than sediment remediation we conclude that the observed ‘improvements’ in the Bovington Stream are in large part due to the sediment control measures (sediment trapping and diversion) implemented since 1998. The project has shown how relatively simple sediment control strategies can improve ecological quality and has provided information on long-term faunal responses which can help managers plan the extent and timing of effective mitigation measures.

1. Improved water quality, through a reduction in diffuse pollution from agricultural sources, is an expected benefit of agri-environment schemes, but this has yet to be demonstrated in practice. Here, we evaluate the impact of Welsh agri-environment schemes on water quality and freshwater ecosystem condition through a combined monitoring and modelling framework.2. To determine the influence of the agri-environment schemes on ecosystem condition, spatially independent catchments dominated by a single scheme (>40% of catchment) were compared to control catchments dominated (>70%) by agricultural land that was not part of any scheme. Biological indicators of water quality were monitored at the outfall of each catchment and a spatially explicit modelling framework of diffuse pollutant emissions applied to each of the 80 catchments.3. Direct comparison ( scheme/non-scheme) was unable to identify any significant effect of agri-environment scheme participation. However, derived biological indicators that reflected organic pollution, eutrophication and pesticide run-off were strongly correlated with modelled concentrations of corresponding diffuse pollutants, thus providing a ground-truth for the models. Scenarios that assessed the correct counterfactuals ( i.e. the influence of scheme entry on pollutant output) were developed for the whole of Wales. The models indicated an important effect of scheme entry on water quality, but this effect was not evenly distributed across the landscape.4. Synthesis and applications. Our results indicate that agri-environment schemes can deliver improvements in water quality, through a reduction in diffuse pollution from agricultural sources. However, it is not easy to demonstrate scheme effectiveness; the combination of field survey and modelling used here provides a framework for addressing these difficulties. A spatially targeted approach for agri-environment scheme options to protect water resources from diffuse pollution is likely to be most effective at delivering water quality improvements.

Excessive delivery of fine sediments to water bodies has a detrimental impact on the biotic elements used for water body status classification. Although diatoms are typically used to assess stress from eutrophication, as fine sediment has the potential to impact diatoms in many ways, it is not surprising that an index based on benthic diatom assemblages has been proposed: the relative abundance of motile species. This measure is based on the fact that many raphid diatom species are capable of migrating through deposited sediment to avoid negative impacts. However, the use of such an index has yet to be fully tested.Various data analysis techniques were used to explore how indices based on diatom assemblages (related to eutrophication and siltation), diatom species, the traits motility, and nutrient affinity responded to a gradient of percentage cover of fine sediment. Although diatom species showed marked variation in their affinity for percentage cover of fine sediment, the relationship between motility (both percent motile and the trait motility) and deposited fine sediment is not sufficiently strong to be used as a reliable indicator of fine sediment stress. We present an approach, which could potentially be used to develop a new index (diatom indicator of sediment conditions) on the basis of the response of diatoms to fine sediment, but caution that this index requires further development before use. Despite the hydromorphology having considerable potential to affect benthic diatoms, existing indices designed to assess eutrophication were robust to hydromorphological modification, reducing the possibility of false diagnosis of impacts.

Excessive inputs of fine‐grained sediment can damage aquatic ecosystems both by degrading habitat condition and by directly impairing biota. Recent research has improved our understanding of how benthic macroinvertebrates respond to fine‐grained sediment stress, leading to the development of a variety of bioassessment indices based on changes in taxonomic composition and biological trait composition. Use of biological traits as indicators of stress has been advocated on the basis of a better mechanistic understanding of the biotic and abiotic factors acting on benthic communities. We quantified changes in the macroinvertebrate biological trait assemblage from a large number of river reaches spanning a national‐scale gradient of increasing agricultural fine sediment delivery and retention, having first factored out variation associated with the natural environmental gradient, with the aim of robustly testing predictions of trait response. We found strong support for 2 of 18 predictions of how macroinvertebrate traits would respond to fine sediment stress. Furthermore, using an independent dataset, we were able to confirm the response of 5 of 6 trait classes that partial RLQ‐fourth corner analysis found to be significantly associated with the fine sediment gradient. Prevalence of eggs as a resistant form, in combination with either an adult aquatic life stage or crawling, provided the best indication of fine sediment conditions in streams, approaching the performance of taxonomic composition‐based sediment indices, CoFSIsp and EPSImtl. This study has robustly confirmed the potential of macroinvertebrate biological traits as indicators of fine sediment impacts.

Excessive sediment pressure on aquatic habitats is of global concern. A unique dataset, comprising instantaneous measurements of deposited fine sediment in 230 agricultural streams across England and Wales, was analysed in relation to 20 potential explanatory catchment and channel variables. The most effective explanatory variable for the amount of deposited sediment was found to be stream power, calculated for bankfull flow and used to index the capacity of the stream to transport sediment. Both stream power and velocity category were highly significant (p ≪ 0.001), explaining some 57% variation in total fine sediment mass. Modelled sediment pressure, predominantly from agriculture, was marginally significant (p < 0.05) and explained a further 1% variation. The relationship was slightly stronger for erosional zones, providing 62% explanation overall. In the case of the deposited surface drape, stream power was again found to be the most effective explanatory variable (p < 0.001) but velocity category, baseflow index and modelled sediment pressure were all significant (p < 0.01); each provided an additional 2% explanation to an overall 50%. It is suggested that, in general, the study sites were transport-limited and the majority of stream beds were saturated by fine sediment. For sites below saturation, the upper envelope of measured fine sediment mass increased with modelled sediment pressure. The practical implications of these findings are that (i) targets for fine sediment loads need to take into account the ability of streams to transport/retain fine sediment, and (ii) where agricultural mitigation measures are implemented to reduce delivery of sediment, river management to mobilise/remove fines may also be needed in order to effect an improvement in ecological status in cases where streams are already saturated with fines and unlikely to self-cleanse.

Summary Detrimental impacts of excessive fine‐grained sediment inputs to streams and rivers are well established. What is less well understood is the susceptibility of different elements of the freshwater biota to such perturbations and how such knowledge of their susceptibility could aid in identifying where excessive fine‐grained sediment is impairing ecological condition. Following the collection of biological and sediment data from 179 streams across England and Wales, representative of a range of river types over a gradient of fine sediment loading, objective statistical approaches were applied to establish relationships between the macroinvertebrate assemblage and fine‐grained sediment inputs to river channels. Having factored out that portion of the biological variation associated with natural environmental gradients, a model comprising mass of organic sediment in erosional areas of the stream bed [predominantly associated with the first axis of the partial canonical correspondence analysis (pCCA)], and mass of fine‐grained sediment in the surface drape of depositional areas and % organic content in erosional areas (associated with the second axis of the pCCA) as explanatory variables best accounted for the residual variation in the macroinvertebrate assemblage. The relative position of taxa along both axes of the pCCA, provided a ranking of taxa in relation to the two gradients of fine‐grained sediment and provided the basis for a new empirically derived diagnostic index for fine‐grained sediment stress in rivers. Two sub‐indices were derived to capture the assemblage responses to both the gradient of organic sediment in erosional areas and the gradient of total fines in depositional areas. The two sub‐indices were then combined to derive the new combined fine sediment index (CoFSIsp). The index was tested on an independent test data set (comprising 127 samples from 83 sites) and was found to provide a robust indication of benthic fine‐grained sediment conditions (Spearman's rank correlations ρ = −0.519 to −0.703). The strength of correlation with the total fine‐grained sediment gradient was always greater than that for other routinely used indices, confirming that CoFSIsp offered additional explanatory power when assessing this stressor of aquatic environments.

Due to a combination of local methanogenesis and high background concentrations in the groundwater, water in the River Lambourn is 51 times supersaturated with methane (162 nmol CH4 L−1). Pore‐water concentrations of methane in the gravels of the riverbed were much lower throughout the year (71 nmol CH4 L−1), suggesting significant methane oxidation. To investigate the potential for methane oxidation as a novel chemosynthetic source of carbon to the food web, we made simultaneous measurements, in laboratory chambers, of primary production, respiration, and methane oxidation associated with the gravels. Biomass‐specific net primary production was up to 2.7 µmol O2 mg−1 chlorophyll (Chl) h−1 and was similarly high for respiration (2.7 µmmol O2 mg−1 Chl h−1). We also found active methane (CH4) oxidation with the rate increasing in proportion to concentration. At the maximum rate of 0.18 µmol CH4 mg−1 Chl h−1 and a growth efficiency of 0.8, net carbon fixation via methane oxidation was equivalent to 6% of the carbon fixed via net photosynthetic primary production. However, production via methane oxidation could be proportionately much greater under the shade of the profuse instream or riparian vegetation, deep in the gravels, and especially during winter, when light is limiting (< 25 µmol quanta m−2 s−1).

We measured the δ13C values of dominant primary consumers and their potential food sources in a groundwater‐fed lowland river. The δ13C of most consumers, such as Gammarus and Simulium, reflected that of the dominant forms of photosynthetic production, whereas the cased larvae of two caddis flies (Agapetus and Silo) were consistently 13C depleted (mean δ13C: ‐41.2% and ‐40.4%, respectively) throughout the year. The river water was supersaturated (approximately 50 times atmospheric) with methane, reflecting both supersaturation in the groundwater and local production in fine sediments. We measured appreciable rates of methane oxidation, relative to water only controls, in the biofilms on gravel, on the caddis fly cases, and on the bottom of larger rocks. In addition, there was a marked difference in the ratio of methane‐oxidizing potential to chlorophyll across those substrata. This ratio was below detection in the biofilm (i.e., no methane oxidation) on the tops of rocks, greater on the bottom of rocks, and maximal for the gravels and the caddis cases. If the caddis larvae acquire most of their carbon by grazing the tops of such rocks (where they are normally found), then they must acquire their depleted δ13C values by occasionally grazing biofilm where the ratio of methane oxidation to chlorophyll was much greater, and the most likely candidate is from their own or conspecific cases. Grazing methane‐oxidizing bacteria could provide the caddis larvae with up to 30% of their carbon, which could represent a true subsidy from an ancient groundwater source.

Background to research The Regulatory Agencies in the UK (the Environment Agency; Scottish Environment Protection Agency; and the Environment & Heritage Service) currently use RIVPACS III+ software to classify the ecological quality of rivers. However, because RIVPACS III+ pre-dates the WFD, there has been a requirement to ensure that the RIVPACS reference sites are fully WFD compliant, to add new biotic indices to the RIVPACS models, and to improve the robustness of the RIVPACS software to fully meet the needs of the Agencies in their delivery of WFD monitoring. These issues have been addressed in this project and have led to the development of new RIVPACS IV predictive models that will be programmed into a new River Invertebrate Classification Tool being built by SEPA. This new system will be based on a modern software programming language, be compatible with the agencies’ computer systems and include the ability to predict new biological indices, produce biological status assessments based on these new indices and be able to estimate the errors involved in using these new indices. Because access to the new system will be essential for the UK Agencies to be able to implementation the WFD, the new tool will be readily and freely available to anyone who might seek to use it. Objectives of research • The overall objective of the project was to produce a new set of RIVPACS predictive models for use within a new River Invertebrate Classification Tool that will be used to classify the ecological status of rivers for Water Framework Directive compliance monitoring • The new RIVPACS models constructed with this project required considerably enhanced functionality compared to RIVPACS III+ to properly address the monitoring requirements of the UK Agencies in their implementation of the Water Framework Directive. Key findings and recommendations This project has produced new RIVPACS IV models with considerably enhanced functionality compared to RIVPACS III+. These models incorporate: • A full revision of the taxonomic framework of RIVPACS to bring the taxonomy up-to-date and enable compatiability across the revised Miatland, Furse code and National Biodiversity Network taxon coding systems used across the UK Agencies and beyond • Predictions that fully satisfy the WFD definition of ‘reference condition’ by adjusting predictions for certain stream types and by removal of sites that were not in reference condition when sampled • Allocation of actual abundance values to family level records in the RIVPACS reference data set. Lack of actual abundance data, especially at family level, has affected all versions of RIVPACS and has constrained the types of biotic indices that RIVPACS can predict • Extension to the suite of biotic indices so that the new system can predict a wider range of reference state “expected” index values. This enables full WFD quality reporting capabilities as well as providing the system with the general functionality to predict a much wider range of indices e.g. intercalibration indices (e.g. ICMi), stress-specific indices, and ecological and functional trait indices • Extension of the uncertainty/errors module to estimate and assess uncertainty in (i) assignment to status class and (ii) comparison of samples for temporal change in quality and status. This needs to be done for a wider range of biotic indices (including those incorporating abundance data) These new RIVPACS IV models can be used by the UK Agencies across Great Britain and Northern Ireland in their WFD compliance monitoring. All of the algorithms, variables and data necessary to build these models have been provided to SEPA for programming into a new River Invertebrate Classification Tool that will be disseminated made free of charge to all interested users