WP1 – Detection and quantifications of emerging contaminants
WP1 will focus on the identification of pollutants due to human pressure (tourism in particular), which may be dangerous for the mountain aquatic organisms. A numerical index will be used to identify potentially dangerous molecules and propose a theoretical classification of toxicity. The algorithm underlying classification indices combines information on the consumption / use of the chemicals, their toxicity on different aquatic organisms and on their dispersion and environmental persistence. The result of the index, which is the list of hazards of chemicals, will be used for planning the subsequent monitoring of EC waters of the Noce River. Following the approach of the European guidelines for the characterization of the ecotoxicological risk, the data generated will be used to evaluate the real level of risk and for new guidelines to reduce the impact on the ecosystem safeguarding the local productive activities.
WP2 – Metagenomic analysis of the microbial communities exposed to emerging contaminants
The WP2 will study the microbial communities as bioindicators of pollution by emerging contaminants (EC) identified in WP1. Automated fingerprinting analyses will be performed to study the genetic structure of the fluvial bacterial and fungal communities in relation to spatial and temporal changes in environmental contamination. The diversity of microbial communities will be analyzed by environmental metagenomic analysis. Bioinformatics analyses will be useful to detect the presence of pathogenic microorganisms and genetic changes of microbial communities in relation to levels of contamination, the functional annotation of genes in order to reconstruct the metabolic pathways, the identification of genes associated with resistance to EC. Metabolic and catabolic genes as the genes involved in the cycles of heavy metals, sulfur and nitrogen, the genes for antibiotic resistance, and metals and related integrons will be quantified and used as bioindication of the presence of the EC.
WP3 – Ecotoxicity and genotoxicity assay: screening of risk of emerging contaminants in macroinvertebrate community
WP3 will focus on the assessment of the effects of the EC identified in WP1 on aquatic animal wildlife. The effects of EC, as individual and as mixture, will be evaluated on macroinvertebrates at community and species level, contributing to the identification of biomarkers useful in the monitoring of EC contamination in rivers. We will study the structure and functional characteristics of the macroinvertebrate community of the selected river reaches, in different seasons, by collecting samples and their subsequent quantitative taxonomic analysis. On the basis of the observed structure (richness, diversity, density) and of the functional groups present, target species belonging to different trophic levels will be selected to perform toxicity and genotoxicity tests. These tests will allow to quantify the toxic resistance of the different species, with different roles in the trophic chain, and to establish environmental concentrations limits for their survival.
WP4 – Ecotoxicity and genotoxicity assay: screening of risk of emerging contaminants in amphibians; effects on chytridiomycosis
WP4 will focus on the assessment of EC effects identified in WP1 on some species of amphibians. Eggs will be collected in the Noce River and reared in the laboratory to the adult stage. Genotoxic damage will be assessed at different development stages. WP4 will investigate (using molecular techniques) also the relationship between the incidence of chytridiomycosis in wild adult specimens and the presence of the EC in the water. Overall, the results of toxicity and genotoxicity assays will allow predictions on the conservation state of endangered species of amphibians.
WP5 – Genotoxic damage in human cell lines caused by mixtures of emerging contaminants
WP5 aims at unearthing any possible effect of EC on human cells and identifying possible alterations of normal cellular physiology upon EC exposure. In order to do that, we will perform dedicated assays to establish the level of cytoxicity and genotoxicity induced by EC mixtures on two human cell lines: the HeLa cells of epithelial origin (uterine cancer) and the breast cancer cells MCF7. Upon identification of the most active mixture, we will further analyze the changes induced in cellular transcription and, eventually, translation employing gene expression analyses, biochemical assays and, if needed, Next Generation Sequencing.
WP6 – Communication and dissemination
WP6 is devoted to dissemination activities, communication and public awareness. The project results will be released to the scientific community (congresses, publications, etc.), the stakeholders and citizenship, explaining their applications in health, environment, hospital-pharmaceutical and engineering-hydraulic field. Within the project the partnership will organize workshops for experts on specific aspects, meetings with stakeholders and local administrators, and events for the public, even to collect opinions and open discussions on this topic of great international relevance. The communication program (including the website) will promote the project and enhance the visibility of the partnership and the Caritro Foundation of Trento.