1. 1 CMEP Plenary Meeting - Prague, 30/06/2011 SETAC Europe 21st Annual Meeting, Milan Joint Research Centre Institute for Environment and Sustainability http://ies.jrc.ec.europa.eu/ CMEP Task 3.2 – Effect based monitoring tools “ Omics” technologies towards the water quality risk assessment for environmental monitoring Raquel Negrão Carvalho Teresa Lettieri

2. 2 CMEP Plenary Meeting - Prague, 30/06/2011 SETAC Europe 21st Annual Meeting, Milan Mode of Action (MoA): chemical-specific molecular signature Omics approaches, including DNA microarray are used to determine MoA of priority substances in model organisms of relevant trophic levels Unaffected by treatment Up-regulated by treatment Down-regulated by treatment

3. 3 CMEP Plenary Meeting - Prague, 30/06/2011 SETAC Europe 21st Annual Meeting, Milan Test species At beginning the sequencing was concentrated on pathogens or species relevant for biomedical field Many other organisms relevant in environment field were sequenced Caulobacter crescentus (Bacteria) Sea squirt (Eukaryota) Fugu fish Diatom (Eukaryota) Daphnia pulex (Eukaryota) rainbow trout Zebrafish fathead minnow Chlamydomonas (green algae) Synechoccocus (cyanobacteria)

4. 4 CMEP Plenary Meeting - Prague, 30/06/2011 SETAC Europe 21st Annual Meeting, Milan Whole systems approaches to environmental assessment Perturbation Pollutants Physical stressors Locations OMICS Metabolome Proteome Transcriptome Interactions Organism Acute Endpoints (EC/LC50) Survival Reproduction Development Available bioassays Vitelogenin CYP450 Androgen receptor Estrogen receptor … Pathway inference Co-regulation discovery MoA Cellular reporters/ Biomarkers Bioassay Identify hazard Classes of pollutants

5. 5 CMEP Plenary Meeting - Prague, 30/06/2011 SETAC Europe 21st Annual Meeting, Milan Molecular based bioassay Saccharomyces cerevisiae Unicellular eukaryote Highly conserved with larger eukaryotes including mammals Not pathogenic easy to handle Does not require any biosafety level Cytotoxicity analysis (selected compounds ) Up regulated Down regulated No change Selection of the mutant strain More sensitivity to the specific class of compounds. Easy approach to generate mutants in the same strain to target different compounds

6. 6 CMEP Plenary Meeting - Prague, 30/06/2011 SETAC Europe 21st Annual Meeting, Milan 1μM Flu. Molecular-based Bioassay Wild-type S288c W303.1a RAD1 YGR131w Control1 μM Flu. 10 3 10 5 10 4 10 2 1010 3 10 5 10 4 10 2 10 3μM Flu. 10 3 10 5 10 4 10 2 10 Development of more sensitive strains e.g. Knock-out of genes involved in Oxidative stress response

7. 7 CMEP Plenary Meeting - Prague, 30/06/2011 SETAC Europe 21st Annual Meeting, Milan - Molecular level response could be linked to a dose-response and allow sample classification regarding environmental hazard - Omics approaches allow exposure of environmental sample without pre- treatment/ fractionation steps - reduced costs, assessment of chemical mixtures and new substances - Different models could be used by different laboratories (if MoA known and biomarkers have been validated for that system) - Information on MoA allows the development of even more sensitive tools FINAL REMARKS

8. 8 CMEP Plenary Meeting - Prague, 30/06/2011 SETAC Europe 21st Annual Meeting, Milan Thank you!