Projects

WP 1 first aims at identifying and quantifying relevant test substances from the groups of pharma-ceuticals (antidiabetics and neuroactive drugs such as selective serotonin reuptake inhibitors) and food additives (artificial sweeteners) in surface waters partly using existing expertise. The focus is on substances of high consumption and environmental relevance. Based on this analysis, the final test compounds will be defined (e.g. metformin, fluoxetine and sucralose), and specific methods for
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The objectives of WP 10 are to prepare the ground for WP 11, which seeks to assess how the key actors perceive biological hazards in aquatic ecosystems. Before these insights can be provided – and guidelines for changing the actors’ perceptions and actions produced –, the actors involved in the evaluation of biological hazards in aquatic ecosystems must be identified, which represents the objective of this WP. The proceeding outline here corresponds to the approach developed by Lang and Tosun
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This WP aims at understanding the awareness of the different actor groups to the impact of two types of substances on aquatic ecosystems in terms of biodiversity and human health: pharmaceu-ticals and food additives. The WP brings together both quantitative and qualitative methods for gen-erating and analyzing data. Each research step will be informed by the findings of the science teams and make an effort to integrate as much of the knowledge produced with regard to toxicolog-ical and ecological
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WP 2 will address metabolites and transformation products (TPs) of compounds selected in WP 1. These will be generated and identified by means of electrochemical synthesis mimicking mainly Phase I metabolism and mass spectrometric detection. The mixture of metabolites and TPs will be used for ligand screening with nuclear receptors (WP 3) using the analytical methodology devel-oped as well as for further toxicity screening for effects on cells (WP 4) and in biota (WPs 5 and 6).

This WP will provide data for receptor binding of the tested xenobiotics at a molecular level in cell-free systems, since in most cases the binding of a compound to a receptor molecule implicates a biological effect, either agonistic or antagonistic. However, since biological approaches are not ca-pable of identifying the chemical structures of environmental contaminants, feasible combinations of bioassays and chemical analysis will be developed and tested. As a pretreatment step concepts of an
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The purpose of WP 4 is to provide a set of toxicological base data on the test substances at the mo-lecular and cellular levels. The focus will not be restricted to basic acute toxicity, but will also cover specific endpoints such as interaction with DNA and DNA damage, the induction of cytochrome P450 as well as the response to glucocorticoid stress hormones and metabolic signaling by glucose. With respect to the selection of test compounds, WP 4 depends on the outcome of WPs 1 and 2.

As an intermediate stage of complexity, fish embryos will be used as a tool to mediate the transfer of data compiled at the molecular and cellular levels (WPs 3 and 4) to intact, fully developed fish (WP 6). To this end, similar endpoints as in WP 4 will be recorded in modified zebrafish embryo assays [85]. In addition, extension of the test duration will allow adding of teratogenic effects to the list of endpoints.

In contrast to WP 5, which addresses in vivo effects in early developmental stages of zebrafish as an established model organism in ecotoxicology, WP 6 will use one indigenous fish species and one species representative of invertebrates in order to verify and, thus, validate the data obtained with zebrafish and also those generated at the in vitro levels (WPs 3 and 4) with respect to their ecologi-cal relevance. Possible adverse effects of the test substances identified in WP 1 will be assessed
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Given that the ultimate goal of ecotoxicology is the identification of effects at the population and ecosystem levels, microbial communities were picked for reasons of their supreme importance in the environment. Two microbial communities (intestinal microbiomes and environmental microbial biofilms) will be investigated with respect to effects by the test substances (cf. WPs 1 and 2). The intestinal biofilm population shifts may be responsible for the health status of the aquatic organisms, whereas
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Results of the multi-level scale approach on effects for the selected micropollutants and their TPs and metabolites will be evaluated and integrated as a base for the creation of a tool box for the ef-fect-based risk assessment. Results will stimulate further work in the development of models linking exposure and effect data with hydrological models. This will require the correlation of results from receptor assays, cell lines and animal models. Microbial data will be linked to the health status
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All data from chemical and biological analyses will be used as sources of data for WP 9. It is not the purpose of Eff-Net to provide a tool to fully model the effects of carbohydrate metabolism-related and neurotoxic substances. Rather, in the sense of an Adverse Outcome Pathway (AOP [33]), the data pool will be analyzed for the suitability of lower scale data (e.g. receptor assay data) to predict the adverse outcome of the test substances at higher levels of organization (e.g. fish). In addition,
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