Spin-Off of Polytechnic University of Marche

1. Spin-Off of Polytechnic University of Marche Environmental Research and Development of Eco-compatible technologies ULIXES kick-off meeting Milan, 28th February 2011 www.ecots.it; www.ecotechsystems.it e-mail: info@ecotechsystems.it Tel/Fax: +39 071 204903 ISO 9001:2008

2. PROFILE Founded in 2003 as Spin-off of Polytechnic University of Marche by marine biologists and ecologists of Department of Marine Sciences (DiSMar) EcoTechSystems L.t.d. provides: • consultancy and services to privates companies for monitoring and management of marine environments • research and development of ecotechnologies for restoration of polluted sites. EcoTechSystems is located at Ancona (Italy) is supported for laboratory analyses by the DiSMar.

3. ULIXES • EcoTechSystems is partner 11 whitin ULIXES and is involved in the following Work Packages: • WP2 - Sampling and characterization of the Mediterranean Sea sites (2 MM) • WP3 - Microbial diversity exploration in the Mediterranean Sea sites (6 MM) • WP4 - Establishment of collections of microbial isolates and metabolites (17 MM) • WP7 - Establishing new product/processes for bioremediation (19 MM) • WP8 - Testing new product/processes by pilot bioremediation treatments in the field (3 MM)

4. ULIXES SCIENTIFIC RATIONALE OF ETS RESEARCH ACTIVITIES Contamination of marine sediments by petroleum hydrocarbons – a widespread problem of coastal anthropized areas Needing of environmental-friendly technologies of bioremediation able to reduce contaminant concentrations to threshold levels Biodegradation of hydrocarbons may be accelerated by enhancing biomass and/or activity of hydrocarbon-degrading microorganisms through biostimulation as well as bioaugmentation strategies Oxygen and nutrient availability are generally assumed limiting factors influencing the rates of biodegradation of hydrocarbons

5. ULIXES Biodegradation of hydrocarbons in the sediment can efficiently occur also in anaerobic conditions mainly due to sulfate and metal (Fe3+/Mn4+) reducers The relatively high biodegradation rates reported in anoxic conditions open new perspectives for the in situ treatment of contaminated sediments where reducing conditions below the sediment surface limit the usefulness of O2 as an electron acceptor which could be supplied to stimulate the degradation of petroleum hydrocarbons Information on how to enhance microbial growth and biodegradation performance in anoxic marine sediments is still very limited

6. ULIXES Marine sediments subjected to strong anthropogenic inputs are often characterized not only by high concentrations of petroleum hydrocarbons, but also by high heavy metal contents The mobility of heavy metals in the sediments depends upon their oxidation-reduction state and their repartition in the different geochemical phases Microbial processes can either increase or decrease heavy metal mobility The bioremediation of marine sediments contaminated by organic and inorganic pollutants should not only identify the best conditions for increasing the biodegradation yields of organic xenobiotics, but also, assess the potential risks associated to changes in heavy metal speciation

7. ULIXES KEY QUESTIONS: Which are the most effective manipulative conditions able to maximize the biodegradation performance of hydrocarbons and the bio-immobilization of metals? Do different metals undergo to different fate (increase/decrease of the mobility) during bio-treatments? Do the geochemical and biological characteristics of the sediments influence the efficiency of the bio-treatments? Which are the relationships between changes of prokaryotic metabolism and diversity induced by bio-treatments and the performance of hydrocarbon degradation and heavy metal mobility?

8. ULIXES • WP2 - Sampling and characterization of the Mediterranean Sea sites • Deliverables: • D2.1 (Report on the geochemical characterization of the sites) – Month 20 • D 2.2 (Report on the overall sampling activity) – Month 30. • Milestones: • MS3 (Achievement of coastal samples) – Month 6 • MS10 (Geochemical characterization of the sites) – Month 20 • MS14 (Achievement of deep sea samples) – Month 26

9. ULIXES Specific activities of ETS within WP2 Two-three different contaminated sites have been already identified: Ancona, Livorno and Piombinoharbours These sites have been selected because background information is already available and because sediments are characterized by different contamination levels as well as typology of contaminants Analyses of organic pollutants (with special focus on hydrocarbons) and heavy metals including their repartition in the different geochemical phases, grain size, mineralogical composition, organic matter content These data, together with the study of microbial biodiversity (WP3), will provide background information for the isolation of bacterial strains which can be involved in the bio-immobilization of metals (WP4) and for the bioremediation experiments which will be carried out in the framework of (WP7).

10. ULIXES • WP3 - Microbial diversity exploration in the Mediterranean Sea sites • Deliverables: • D3.1 (Diversity database of microbial species present in the different environments) – Month 15 • D 3.2 (Diversity database of metabolic functions characterizing the polluted environments) – Month 20. • D3.4 (Microbial biodiversity map) – Month 36 • Milestones: • MS 37 (Definition of the microbial species diversity) – Month 15 • MS11 (Definition of the metabolic function diversity) – Month 20

11. ULIXES • Specific activities of ETS within WP3 • The information on chemical contamination acquired in WP2 will be coupled with information on abundance, biomass and biodiversity of the microbial benthic community. • Total prokaryotic abundance and biomass • Physical-chemical cell detachment , staining and counts under epifluorescence microscopy • Prokaryotic biodiversity • T-RFLP and ARISA analyses coupled with gene cloning and sequences and/or 454 pyrosequencing

12. ULIXES • WP4 – Establishment of collections of microbial isolates and metabolites • Deliverable: • D 4.4. Establishment of collection 4 • METAL BIOSTABILIZING PROKARYOTES – Month 24. ETS Lead beneficiary • ETS will provide a collection of microbial strains (both aerobic and anaerobic) with can decrease the mobility of heavy metals thorough specific metabolic reactions (e.g. formation of metal-sulphide complexes) and/or extracellular compounds produced during their growth (exopolysaccarides). Subcultures and Identification of microbial strains Test for effectiveness Cultures and screening

13. ULIXES • WP7 - Establishing new product/processes for bioremediation • Deliverables: • D 7.1 (Establishment of a metal/metalloids fate model in sediments) – Month 28. ETS Lead beneficiary • Milestones: • MS15 (Optimization of the novel biodegradation processes) – Month 35. • ETS will perform specific bioremediation experiments in oxic and anoxic conditions to identify potential relationships between changes in prokaryotic growth and biodiversity, biodegradation potential of hydrocarbons and effects on the mobility of metals and/or metalloids. • Microcosm experiments: anoxia induction, addition of SRB strains, sodium acetate addition (as electron donor for stimulating sulfate-reducing processes), inorganic nutrients addition, changing of physico-chemical conditions

14. ULIXES • WP8 - Testing new product/processes by pilot bioremediation treatments in the field • Mesocosm experiments in coastal sites used for sampling activities • Treatments to be planned on basis of results coming from WP7 • Time series incubations (1 week or more) • Regular monitoring of environmental and microbial parameters