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AbstractCentral European riverine networks are subject to widely varying local anthropogenic pressures, forcing species with limited dispersal abilities to adapt or become locally extinct. Previous catchment-wide studies have shown that some invertebrates tend to have pronounced population structuring throughout mountainous river networks, raising the question of whether this also translates into small-scale phenotypic differentiation and adaptation to local stressors. One such species is the headwater crustacean species Gammarus fossarum clade 11 (or lineage B), which we restudied in terms of population structure four years after first assessment. Our aim was not only to document the temporal stability/dynamics of the population structure, but we asked whether a small-scale genetic structuring also results in phenotypic differentiation and different susceptibility to a commonly applied pesticide. Therefore, we re-assessed population structure based on COI haplotypes and their frequencies, and quantified key parameters related to morphological and life-history differentiation. Furthermore, we examined the difference in sensitivity towards the pyrethroid insecticide deltamethrin. COI haplotype patterns were found to be stable over time and confirmed the small-scale population structuring within the catchment, with isolated headwater populations and connected downstream populations. While little life-history differentiation was observed, marked differences in susceptibility to the pyrethroid insecticide were found. Populations from pristine sites responded significantly more tolerant than populations from anthropogenically impacted sites—showing that prior exposure to a spectrum of stressors does not automatically increase tolerance to a specific stressor. Therefore, our study demonstrates that limited dispersal capacity is reflected not only in population structure, but also in small-scale variation in susceptibility to anthropogenic disturbance. The system thus provides a suitable experimental landscape to test the impact of further stressors (e.g., other novel entities, including pesticides with other modes of action) on locally isolated populations. Based on these findings, important recommendations for the protection of riverine species and their intraspecific genetic variation can be developed.

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.

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.

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.

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.

24 páginas, 2 figuras, 1 tabla. ; 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. ; Peer reviewed

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.