Maximising environmental pressure-response relationship signals from diatom-based metabarcoding in rivers.

DNA metabarcoding has been performed on a large number of river phytobenthos samples collected from the UK, using rbcL primers optimised for diatoms. Within this dataset the composition of non-diatom sequence reads was studied and the effect of including these in models for evaluating the nutrient gradient was assessed. Whilst many non-diatom taxonomic groups were detected, few contained the full diversity expected in riverine environments.

Setting nutrient boundaries to protect aquatic communities: The importance of comparing observed and predicted classifications using measures derived from a confusion matrix.

Defining nutrient thresholds that protect and support the ecological integrity of aquatic ecosystems is a fundamental step in maintaining their natural biodiversity and preserving their resilience. With increasing catchment pressures and climate change, it is more important than ever to develop clear methods to establish thresholds for status classification and management of waters. This must often be achieved using complex data and should be robust to interference from additional pressures as well as ameliorating or confounding conditions.

Estimating nutrient thresholds for eutrophication management: Novel insights from understudied lake types.

Nutrient targets based on pressure-response models are essential for defining ambitions and managing eutrophication. However, the scale of biogeographical variation in these pressure-response relationships is poorly understood, which may hinder eutrophication management in regions where lake ecology is less intensively studied. In this study, we derive ecology-based nutrient targets for five major ecoregions of Europe: Northern, Central-Baltic, Alpine, Mediterranean and Eastern Continental.

Establishing ecologically-relevant nutrient thresholds: A tool-kit with guidance on its use.

One key component of any eutrophication management strategy is establishment of realistic thresholds above which negative impacts become significant and provision of ecosystem services is threatened. This paper introduces a toolkit of statistical approaches with which such thresholds can be set, explaining their rationale and situations under which each is effective. All methods assume a causal relationship between nutrients and biota, but we also recognise that nutrients rarely act in isolation.

Diatom DNA metabarcoding for ecological assessment: Comparison among bioinformatics pipelines used in six European countries reveals the need for standardization.

Ecological assessment of lakes and rivers using benthic diatom assemblages currently requires considerable taxonomic expertise to identify species using light microscopy. This traditional approach is also time-consuming. Diatom metabarcoding is a promising alternative and there is increasing interest in using this approach for routine assessment.

European aquatic ecological assessment methods: A critical review of their sensitivity to key pressures.

The European Union has embarked on a policy which aims to achieve good ecological status in all surface waters (i.e. rivers, lakes, transitional and coastal waters).

Executing multi-taxa eDNA ecological assessment via traditional metrics and interactive networks.

Current approaches to ecological assessment are limited by the traditional morpho-taxonomic methods presently employed and the inability to meet increasing demands for rapid assessments. Advancements in high throughput sequencing now enable rapid high-resolution ecological assessment using environmental DNA (eDNA). Here we test the ability of using eDNA-based ecological assessment methods against traditional assessment of two key indicator groups (diatoms and macroinvertebrates) and show how eDNA across multiple gene regions (COI, rbcL, 12S and 18S) can be used to infer interactive networks that link to ecological assessment criteria.

Nutrient criteria for surface waters under the European Water Framework Directive: Current state-of-the-art, challenges and future outlook.

The aim of European water policy is to achieve good ecological status in all rivers, lakes, coastal and transitional waters by 2027. Currently, more than half of water bodies are in a degraded condition and nutrient enrichment is one of the main culprits. Therefore, there is a pressing need to establish reliable and comparable nutrient criteria that are consistent with good ecological status.

Diat.barcode, an open-access curated barcode library for diatoms.

Diatoms (Bacillariophyta) are ubiquitous microalgae which produce a siliceous exoskeleton and which make a major contribution to the productivity of oceans and freshwaters. They display a huge diversity, which makes them excellent ecological indicators of aquatic ecosystems. Usually, diatoms are identified using characteristics of their exoskeleton morphology.

Establishing nutrient thresholds in the face of uncertainty and multiple stressors: A comparison of approaches using simulated datasets.

Various methods have been proposed to identify threshold concentrations of nutrients that would support good ecological status, but the performance of these methods and the influence of other stressors on the underlying models have not been fully evaluated. We used synthetic datasets to compare the performance of ordinary least squares, logistic and quantile regression, as well as, categorical methods based on the distribution of nutrient concentrations categorised by biological status. The synthetic datasets used differed in their levels of variation between explanatory and response variables, and were centered at different positions along the stressor (nutrient) gradient.

Connecting the morphological and molecular species concepts to facilitate species identification within the genus Fragilaria (Bacillariophyta).

This paper explores the diversity and taxonomy of species within Fragilaria sensu stricto, an abundant and ecologically important diatom genus, taking advantage of cultured and DNA-barcoded material. The goal is to facilitate the identification of European taxa within this complex, providing a unified view on morphological and molecular diversity. There is a general agreement that the separation of species within the group of Fragilaria is difficult because morphological descriptions of species are not consistent between authorities, ongoing taxonomic revisions have resulted in species described with standards of the late 20th and 21st centuries alongside descriptions based on 19th century (light microscopical) criteria, and because not all diagnostic characters can be seen in all specimens encountered in routine analyses.

Protecting and restoring Europe's waters: An analysis of the future development needs of the Water Framework Directive.

The Water Framework Directive (WFD) is a pioneering piece of legislation that aims to protect and enhance aquatic ecosystems and promote sustainable water use across Europe. There is growing concern that the objective of good status, or higher, in all EU waters by 2027 is a long way from being achieved in many countries. Through questionnaire analysis of almost 100 experts, we provide recommendations to enhance WFD monitoring and assessment systems, improve programmes of measures and further integrate with other sectoral policies.

Macrophyte assessment in European lakes: Diverse approaches but convergent views of 'good' ecological status.

The European Water Framework Directive has been adopted by Member States to assess and manage the ecological integrity of surface waters. Specific challenges include harmonizing diverse assessment systems across Europe, linking ecological assessment to restoration measures and reaching a common view on 'good' ecological status. In this study, nine national macrophyte-based approaches for assessing ecological status were compared and harmonized, using a large dataset of 539 European lakes.

Deriving nutrient criteria to support 'good' ecological status in European lakes: An empirically based approach to linking ecology and management.

European water policy has identified eutrophication as a priority issue for water management. Substantial progress has been made in combating eutrophication but open issues remain, including setting reliable and meaningful nutrient criteria supporting 'good' ecological status of the Water Framework Directive. The paper introduces a novel methodological approach - a set of four different methods - that can be applied to different ecosystems and stressors to derive empirically-based management targets.

Small Water Bodies in Great Britain and Ireland: Ecosystem function, human-generated degradation, and options for restorative action.

Small, 1st and 2nd-order, headwater streams and ponds play essential roles in providing natural flood control, trapping sediments and contaminants, retaining nutrients, and maintaining biological diversity, which extend into downstream reaches, lakes and estuaries. However, the large geographic extent and high connectivity of these small water bodies with the surrounding terrestrial ecosystem makes them particularly vulnerable to growing land-use pressures and environmental change. The greatest pressure on the physical processes in these waters has been their extension and modification for agricultural and forestry drainage, resulting in highly modified discharge and temperature regimes that have implications for flood and drought control further downstream.

Declining glacier cover threatens the biodiversity of alpine river diatom assemblages.

Climate change poses a considerable threat to the biodiversity of high altitude ecosystems worldwide, including cold-water river systems that are responding rapidly to a shrinking cryosphere. Most recent research has demonstrated the severe vulnerability of river invertebrates to glacier retreat but effects upon other aquatic groups remain poorly quantified. Using new data sets from the European Alps, we show significant responses to declining glacier cover for diatoms, which play a critical functional role as freshwater primary producers.

Implementation options for DNA-based identification into ecological status assessment under the European Water Framework Directive.

Assessment of ecological status for the European Water Framework Directive (WFD) is based on "Biological Quality Elements" (BQEs), namely phytoplankton, benthic flora, benthic invertebrates and fish. Morphological identification of these organisms is a time-consuming and expensive procedure. Here, we assess the options for complementing and, perhaps, replacing morphological identification with procedures using eDNA, metabarcoding or similar approaches.

A Water Framework Directive-compatible metric for assessing acidification in UK and Irish rivers using diatoms.

Freshwater acidification continues to be a major problem affecting large areas of Europe, and while there is evidence for chemical recovery, similar evidence for biological recovery of freshwaters is sparse. The need for a methodology to identify waterbodies impacted acidification and to assess the extent of biological recovery is relevant to the EU Water Framework Directive, which requires methods to quantify differences in biology between impacted and unimpacted or reference sites. This study presents a new WFD-compliant metric based on diatoms (Diatom Acidification Metric: DAM) for assessing the acidification status of rivers.

Benthic algal assessment of ecological status in European lakes and rivers: Challenges and opportunities.

This opinion paper introduces a special series of articles dedicated to freshwater benthic algae and their use in assessment and monitoring. This special series was inspired by talks presented at the 9th International Congress on the Use of Algae for Monitoring Rivers and Comparable Habitats (Trento, Italy, 2015), the latest of a series of meetings started in 1991. In this paper, we will first provide a brief overview of phytobenthos methods in Europe.

Redundancy in the ecological assessment of lakes: Are phytoplankton, macrophytes and phytobenthos all necessary?

Although the Water Framework Directive specifies that macrophytes and phytobenthos should be used for the ecological assessment of lakes and rivers, practice varies widely throughout the EU. Most countries have separate methods for macrophytes and phytobenthos in rivers; however, the situation is very different for lakes. Here, 16 countries do not have dedicated phytobenthos methods, some include filamentous algae within macrophyte survey methods whilst others use diatoms as proxies for phytobenthos.

RAPPER: A new method for rapid assessment of macroalgae as a complement to diatom-based assessments of ecological status.

Most methods for ecological assessment developed since the onset of the Water Framework Directive require substantial effort by skilled analysts and are therefore expensive to use. RAPPER ("Rapid Assessment of PeriPhyton Ecology in Rivers") is a high level ecological "triage" method that enables rapid screening of sites within a water body to enable managers to identify areas subject to nutrient pressures. The method involves a survey of macroscopic algae within 10m lengths of watercourses, taking samples for subsequent identification, and assessing cover.

Interactions between pH and nutrients on benthic algae in streams and consequences for ecological status assessment and species richness patterns.

Eutrophication and acidification are among the major stressors on freshwater ecosystems in northern Europe and North America, but possible consequences of interactions between pH and nutrients on ecological status assessment and species richness patterns have not previously been assessed. Using data from 52 river sites throughout Norway, we investigated the combined effects of pH and nutrients on benthic algae assemblages, specifically 1) taxa-specific couplings between nutrient and acidity traits, 2) the degree of consistency between different biotic indices, separately for nutrients and acid conditions, 3) the impact of pH on nutrient indices and phosphorus on indices of acid conditions, and 4) the impact of pH and phosphorus supply on diatom and non-diatom taxon richness. We found that 1) acid-tolerant taxa are generally associated with nutrient-poor conditions, with only a few exceptions; this is probably more a consequence of habitat availability than reflecting true ecological niches; 2) correlation coefficients between nutrient indices and TP, as well as acid conditions indices and pH were barely affected when the confounding factor was removed; 3) the association of acid-tolerant taxa with nutrient-poor conditions means that the lowest possible nutrient index at a site, as indicated by benthic algae, is lower at acid than at circumneutral sites.

The European reference condition concept: A scientific and technical approach to identify minimally-impacted river ecosystems.

One objective of the European Union (EU)'s Water Framework Directive (WFD: Directive 2000/60/EC) is for all European surface waters to achieve 'good status' by 2015. In support of this objective, the EU has facilitated an intercalibration exercise to ensure harmonized definitions of the status of water bodies, reflecting the deviation of their properties (mainly biotic assemblages) from a minimally disturbed state, termed the "reference condition". One of the major challenges of the WFD has been to find common approaches for defining reference conditions and to define the level of anthropogenic intervention allowed in reference sites.

Effect of phosphorus stripping on water chemistry and diatom ecology in an eastern lowland river.

The effect of nutrient removal at Ashford sewage treatment works on the benthic diatom ecology of the River Stour was studied. This paper describes assemblages above and below the works both before and after phosphorus stripping was installed. Taxa typical of eutrophic conditions dominated all samples, including those upstream of the works, suggesting that the river was already eutrophic before receiving the sewage effluent.

Short term dynamics of diatoms in an upland stream and implications for monitoring eutrophication.

A study designed to examine the effect of intermittent diffuse nutrient inputs on diatom assemblages in a small upland stream in northern England was confounded by other changes that occurred in the stream just before fertilisation started. A flora dominated by attached diatoms changed, over a period of about 3 weeks to one dominated by motile Nitzschia spp. In the absence of any likely human causes, the most likely reason for this change was undercutting of a meander upstream, releasing fine silts which favoured motile diatoms.