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The protection, preservation and restoration of aquatic ecosystems and their functions are of global importance. For European states it became legally binding mainly through the EUWater Framework Directive (WFD). In order to assess the ecological status of a given water body, aquatic biodiversity data are obtained and compared to a reference water body. The quantified mismatch obtained determines the extent of potential management actions. The current approach to biodiversity assessment is based on morpho-taxonomy. This approach has many drawbacks such as being time consuming, limited in temporal and spatial resolution, and error-prone due to the varying individual taxonomic expertise of the analysts. Novel genomic tools can overcome many of the aforementioned problems and could complement or even replace traditional bioassessment. Yet, a plethora of approaches are independently developed in different institutions, thereby hampering any concerted routine application. The goal of this Action is to nucleate a group of researchers across disciplines with the task to identify gold-standard genomic tools and novel ecogenomic indices for routine application in biodiversity assessments of European fresh- and marine water bodies. Furthermore, DNAqua-Net will provide a platform for training of the next generation of European researchers preparing them for the new technologies. Jointly with water managers, politicians, and other stakeholders, the group will develop a DNAqua-Net: Developing new genetic tools for bioassessment and monitoring conceptual framework for the standard application of eco-genomic tools as part of legally binding assessments.

Several national and international environmental laws require countries to meet clearly defined targets with respect to the ecological status of aquatic ecosystems. In Europe, the EU-Water Framework Directive (WFD; 2000/60/EC) represents such a detailed piece of legislation. The WFD that requires the European member countries to achieve an at least 'good' ecological status of all surface waters at latest by the year 2027. In order to assess the ecological status of a given water body under the WFD, data on its aquatic biodiversity are obtained and compared to reference status. The mismatch between these two metrics then is used to derive the respective ecological status class. While the workflow to carry out the assessment is well established, it relies only on few biological groups (typically fish, macroinvertebrates and a few algal taxa such as diatoms), is time consuming and remains at a lower taxonomic resolution, so that the identifications can be done routinely by non-experts with an acceptable learning curve. Here, novel genetic and genomic tools provide new solutions to speed up the process and allow to include a much greater proportion of biodiversity in the assessment process. further, results are easily comparable through the genetic 'barcodes' used to identify organisms. The aim of the large international COST Action DNAqua-Net (http://dnaqua.net/) is to develop strategies on how to include novel genetic tools in bioassessment of aquatic ecosystems in Europe and beyond and how to standardize these among the participating countries. It is the ambition of the network to have these new genetic tools accepted in future legal frameworks such as the EU-Water Framework Directive (WFD; 2000/60/EC) and the Marine Strategy Framework Directive (2008/56/EC). However, a prerequisite is that various aspects that start from the validation and completion of DNA Barcode reference databases, to the lab and field protocols, to the analysis processes as well as the subsequently derived biotic indices and metrics are ...

Several national and international environmental laws require countries to meet clearly defined targets with respect to the ecological status of aquatic ecosystems. In Europe, the EU-Water Framework Directive (WFD; 2000/60/EC) represents such a detailed piece of legislation. The WFD that requires the European member countries to achieve an at least 'good' ecological status of all surface waters at latest by the year 2027. In order to assess the ecological status of a given water body under the WFD, data on its aquatic biodiversity are obtained and compared to reference status. The mismatch between these two metrics then is used to derive the respective ecological status class. While the workflow to carry out the assessment is well established, it relies only on few biological groups (typically fish, macroinvertebrates and a few algal taxa such as diatoms), is time consuming and remains at a lower taxonomic resolution, so that the identifications can be done routinely by non-experts with an acceptable learning curve. Here, novel genetic and genomic tools provide new solutions to speed up the process and allow to include a much greater proportion of biodiversity in the assessment process. further, results are easily comparable through the genetic 'barcodes' used to identify organisms. The aim of the large international COST Action DNAqua-Net (http://dnaqua.net/) is to develop strategies on how to include novel genetic tools in bioassessment of aquatic ecosystems in Europe and beyond and how to standardize these among the participating countries. It is the ambition of the network to have these new genetic tools accepted in future legal frameworks such as the EU-Water Framework Directive (WFD; 2000/60/EC) and the Marine Strategy Framework Directive (2008/56/EC). However, a prerequisite is that various aspects that start from the validation and completion of DNA Barcode reference databases, to the lab and field protocols, to the analysis processes as well as the subsequently derived biotic indices and metrics are dealt with and commonly agreed upon. Furthermore, many pragmatic questions such as adequate short and long-term storage of samples or specimens for further processing or to serve as an accessible reference need also be addressed. In Europe the conformity and backward compatibility of the new methods with the existing legislation and workflows are further of high importance. Without rigorous harmonization and inter-calibration concepts, the implementation of the powerful new genetic tools will be substantially delayed in real-world legal framework applications. After a short introduction on the structure and vision of DNAqua-Net, we discuss how the DNAqua-Net community considers possibilities to include novel DNA-based approaches into current bioassessment and how formal standardization e.g. through the framework of CEN (The European Committee for Standardization) may aid in that process (Hering et al. 2018, Leese et al. 2016, Leese et al. 2018. Further we explore how TDWG data standards can further facilitate swift adoption of the genetic methods in routine use. We further present potential impacts of the legislative requirements of the Nagoya Protocol on the exchange of genetic resources and their implications for biomonitoring. Last but not least, we will touch upon the rather unexpected influence that the new General Data Protection Regulation (GDPR) may have on the bioassessment work in practice.

Abstract
Background
Freshwater ecosystem degradation and biodiversity decline are strongly associated with intensive agricultural practices. Simultaneously occurring agricultural stressors can interact in complex ways, preventing an accurate prediction of their combined effects on aquatic biota. Here, we address the limited mechanistic understanding of multiple stressor effects of two globally important stressors, an insecticide (chlorantraniliprole), and increased fine sediment load and assessed their impact on the transcriptomic profile of two stream macroinvertebrates: the amphipod Gammarus pulex and the caddisfly Lepidostoma basale.

Results
We identified mainly antagonistic stressor interactions at the transcriptional level, presumably because the insecticide adsorbed to fine sediment particles. L. basale, which is phylogenetically more closely related to the insecticide's target taxon Lepidoptera, exhibited strong transcriptional changes when the insecticide stressor was applied, whereas no clear response patterns were observed in the amphipod G. pulex. These differences in species vulnerability can presumably be attributed to molecular mechanisms determining the cellular affinity toward a stressor as well as differential exposure patterns resulting from varying ecological requirements between L. basale and G. pulex. Interestingly, the transcriptional response induced by insecticide exposure in L. basale was not associated with a disruption of the calcium homeostasis, which is the described mode of action for chlorantraniliprole. Instead, immune responses and alterations of the developmental program appear to play a more significant role.

Conclusions
Our study shows how transcriptomic data can be used to identify multiple stressor effects and to explore the molecular mechanisms underlying stressor-induced physiological responses. As such, stressor effects assessed at the molecular level can inform about modes of action of chemicals and their interplay with non-chemical stressors. We demonstrated that stressor effects vary between different organismic groups and that insecticide effects are not necessarily covered by their described mode of action, which has important implications for environmental risk assessment of insecticides in non-target organisms.

Abstract
Background
Fungicides are frequently used in agriculture and can enter freshwater ecosystems through multiple pathways. The negative impacts of fungicides on microorganisms, fungi in particular, and their functions such as leaf decomposition have been repeatedly shown. In our previous microcosm experiment with three consecutive cycles of fungicide exposure and colonisation of leaf substrate, we found clear functional changes, but no differences in fungal community structure could be detected using morphological identification by analysing the spores of aquatic hyphomycetes. In this study, we examined the effects on fungal and bacterial community composition in detail using ITS and 16S metabarcoding and comparing the results to morphologically assessed community composition.

Results
While we found fewer species with metabarcoding than with morphological identification, metabarcoding also enabled the identification of several fungal species that were otherwise unidentifiable morphologically. Moreover, by distinguishing individual amplicon sequence variants (ASVs) metabarcoding provided greater taxonomic resolution. In line with the morphological results, metabarcoding neither revealed effects of fungicides on the aquatic hyphomycetes nor on the total fungal or bacterial community composition. However, several ASVs responded significantly to fungicides, demonstrating variable tolerances within species.

Conclusions
Overall, the absence of detectable effects of fungicides on the community structure despite clear functional effects, suggests a complex relationship between community structure and the ecosystem function of leaf decomposition.

The protection, preservation and restoration of aquatic ecosystems and their functions are of global importance. For European states it became legally binding mainly through the EUWater Framework Directive (WFD). In order to assess the ecological status of a given water body, aquatic biodiversity data are obtained and compared to a reference water body. The quantified mismatch obtained determines the extent of potential management actions. The current approach to biodiversity assessment is based on morpho-taxonomy. This approach has many drawbacks such as being time consuming, limited in temporal and spatial resolution, and error-prone due to the varying individual taxonomic expertise of the analysts. Novel genomic tools can overcome many of the aforementioned problems and could complement or even replace traditional bioassessment. Yet, a plethora of approaches are independently developed in different institutions, thereby hampering any concerted routine application. The goal of this Action is to nucleate a group of researchers across disciplines with the task to identify gold-standard genomic tools and novel ecogenomic indices for routine application in biodiversity assessments of European fresh- and marine water bodies. Furthermore, DNAqua-Net will provide a platform for training of the next generation of European researchers preparing them for the new technologies. Jointly with water managers, politicians, and other stakeholders, the group will develop a conceptual framework for the standard application of eco-genomic tools as part of legally binding assessments. ; European Cooperation in Science & Technology program (EU COST) ...

Water and water bodies are vital for people and nature. They provide both important resources and valuable habitats for life. The conservation and use of water systems must hence be reconciled to ensure the best possible path to sustainable development. Pressure on water resources and aquatic ecosystems increases continuously both in Germany and worldwide. Agriculture, industries, the energy and water economy, settlements and traffic, as well as recreation contribute to this development. Climate change, including increases in the frequency and severity of extreme events such as droughts and intense precipitation, exacerbates the situation. Water is becoming scarce for people and ecosystems or is getting out of control during extreme rainfall. Increased damage to infrastructure, water pollution and degraded ecosystems limited in their functionality ensue. Water policy must cope with conflicting goals of water resource use and conservation. The resulting conflicts are serious and complex, and have evident repercussions for practical water management, calling for new approaches to solve the pressing issues. In view of the complex task and rapidly changing framework conditions, a comprehensive understanding of water systems is imperative to implementing viable prevention and adaptation strategies. The topics to consider must range from individual hydrological, ecological and technical processes to system interrelationships and dynamics, and to economic, social and political issues. This breadth is a challenge for water research. Accordingly, the German Water Science Alliance aspires to link fundamental scientific insights across disciplines to practical solutions of water issues with a view to promote evidence-based water policy supporting sustainable water resource and ecosystem management – in Germany, Europe and worldwide. The present framework paper identifies four central thematic challenges along these lines: 1. Hydrological extremes - developing sustainable adaptation options to cope with increasingly ...

The protection, preservation and restoration of aquatic ecosystems and their functions are of global importance. For European states it became legally binding mainly through the EU-Water Framework Directive (WFD). In order to assess the ecological status of a given water body, aquatic biodiversity data are obtained and compared to a reference water body. The quantified mismatch obtained determines the extent of potential management actions. The current approach to biodiversity assessment is based on morpho-taxonomy. This approach has many drawbacks such as being time consuming, limited in temporal and spatial resolution, and error-prone due to the varying individual taxonomic expertise of the analysts. Novel genomic tools can overcome many of the aforementioned problems and could complement or even replace traditional bioassessment. Yet, a plethora of approaches are independently developed in different institutions, thereby hampering any concerted routine application. The goal of this Action is to nucleate a group of researchers across disciplines with the task to identify gold-standard genomic tools and novel eco-genomic indices for routine application in biodiversity assessments of European fresh- and marine water bodies. Furthermore, DNAqua-Net will provide a platform for training of the next generation of European researchers preparing them for the new technologies. Jointly with water managers, politicians, and other stakeholders, the group will develop a conceptual framework for the standard application of eco-genomic tools as part of legally binding assessments. ; © Leese F et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

The protection, preservation and restoration of aquatic ecosystems and their functions are of global importance. For European states it became legally binding mainly through the EU-Water Framework Directive (WFD). In order to assess the ecological status of a given water body, aquatic biodiversity data are obtained and compared to a reference water body. The quantified mismatch obtained determines the extent of potential management actions. The current approach to biodiversity assessment is based on morpho-taxonomy. This approach has many drawbacks such as being time consuming, limited in temporal and spatial resolution, and error-prone due to the varying individual taxonomic expertise of the analysts. Novel genomic tools can overcome many of the aforementioned problems and could complement or even replace traditional bioassessment. Yet, a plethora of approaches are independently developed in different institutions, thereby hampering any concerted routine application. The goal of this Action is to nucleate a group of researchers across disciplines with the task to identify gold-standard genomic tools and novel eco-genomic indices for routine application in biodiversity assessments of European fresh- and marine water bodies. Furthermore, DNAqua-Net will provide a platform for training of the next generation of European researchers preparing them for the new technologies. Jointly with water managers, politicians, and other stakeholders, the group will develop a conceptual framework for the standard application of eco-genomic tools as part of legally binding assessments. ; ISSN:2367-7163

The protection, preservation and restoration of aquatic ecosystems and their functions are of global importance. For European states it became legally binding mainly through the EUWater Framework Directive (WFD). In order to assess the ecological status of a given water body, aquatic biodiversity data are obtained and compared to a reference water body. The quantified mismatch obtained determines the extent of potential management actions. The current approach to biodiversity assessment is based on morpho-taxonomy. This approach has many drawbacks such as being time consuming, limited in temporal and spatial resolution, and error-prone due to the varying individual taxonomic expertise of the analysts. Novel genomic tools can overcome many of the aforementioned problems and could complement or even replace traditional bioassessment. Yet, a plethora of approaches are independently developed in different institutions, thereby hampering any concerted routine application. The goal of this Action is to nucleate a group of researchers across disciplines with the task to identify gold-standard genomic tools and novel ecogenomic indices for routine application in biodiversity assessments of European fresh- and marine water bodies. Furthermore, DNAqua-Net will provide a platform for training of the next generation of European researchers preparing them for the new technologies. Jointly with water managers, politicians, and other stakeholders, the group will develop a conceptual framework for the standard application of eco-genomic tools as part of legally binding assessments. ; Funding program European Cooperation in Science & Technology program (EU COST)

Aquatic biomonitoring has become an essential task in Europe and many other regions as a consequence of strong anthropogenic pressures affecting the health of lakes, rivers, oceans and groundwater. A typical assessment of the environmental quality status, such as it is required by European but also North American and other legislation, relies on matching the composition of assemblages of organisms identified using morphological criteria present in aquatic ecosystems to those expected in the absence of anthropogenic pressures. Through decade-long and difficult intercalibration exercises among networks of regulators and scientists in European countries, a pragmatic biomonitoring approach was developed and adopted, which now produces invaluable information. Nonetheless, this approach is based on several hundred different protocols, making it susceptible to issues with comparability, scale and resolution. Furthermore, data acquisition is often slow due to a lack of taxonomic experts for many taxa and regions and time-consuming morphological identification of organisms. High-throughput genetic screening methods such as (e)DNA metabarcoding have been proposed as a possible solution to these shortcomings. Such "next-generation biomonitoring", also termed "biomonitoring 2.0", has many advantages over the traditional approach in terms of speed, comparability and costs. It also creates the potential to include new bioindicators and thereby further improves the assessment of aquatic ecosystem health. However, several major conceptual and technological challenges still hinder its implementation into legal and regulatory frameworks. Academic scientists sometimes tend to overlook legal or socioeconomic constraints, which regulators have to consider on a regular basis. Moreover, quantification of species abundance or biomass remains a significant bottleneck to releasing the full potential of these approaches. Here, we highlight the main challenges for next-generation aquatic biomonitoring and outline principles and good ...

This report presents the outcome of the joint work of PhD students and senior researchers working with DNA-based biodiversity assessment approaches with the goal to facilitate others the access to definitions and explanations about novel DNA-based methods. The work was performed during a PhD course (SLU PNS0169) at the Swedish University of Agricultural Sciences (SLU) in Uppsala, Sweden. The course was co-organized by the EU COST research network DNAqua-Net and the SLU Research Schools Focus on Soils and Water (FoSW) and Ecology - basics and applications. DNAqua-Net (COST Action CA15219, 2016-2020) is a network connecting researchers, water managers, politicians and other stakeholders with the aim to develop new genetic tools for bioassessment of aquatic ecosystems in Europe and beyond. The PhD course offered a comprehensive overview of the paradigm shift from traditional morphology-based species identification to novel identification approaches based on molecular markers. We covered the use of molecular tools in both basic research and applied use with a focus on aquatic ecosystem assessment, from species collection to the use of diversity in environmental legislation. The focus of the course was on DNA (meta)barcoding and aquatic organisms. The knowledge gained was shared with the general public by creating Wikipedia pages and through this collaborative Open Access publication, co-authored by all course participants.