Seven research and communication based work packages (WPs) of NanoFATE were, in turn, grouped into the three main research components of the project. Two separate work packages (WP 8, WP 9) were dedicated to dissemination and project management and administration. The overall design of the project achieved a consistent flow of data and information to complete an integrated development of tools for assessing the environmental fate of ENPs, their specific toxicological effects and the risks posed. The 9 Project Objectives were mapped onto the WPs, in some cases directly to an individual WP, but in many cases transversally across WPs. With this integration, NanoFATE project results not only provided a template for ENP risk assessment, but also supported technological innovation centered on the eco-responsible design of ENPs and methods for safe ENP disposal/recycling. Work package 1 Characterisation and tracking of ENPs during processes involved in fate and toxicity Objective 1: Design and manufacture of tagged ENPs for tracking in fate and toxicity studies. Direct contributions by work package Objective 3: Analyse ENP interactions with environmental and biological entities using advanced microscope and physical analysis. Direct contributions by work package Objective 4: Study ENP fate and behaviour through wastewater treatment processes and in soils. Direct contributions by work package Objective 6: Establish and model how environment physico-chemical properties in wastewater, natural waters and soil govern ENP parameters that each may ultimately affect bioavailability to organisms. Indirect contributions by work package Objective 7: Establish the mechanisms of uptake, internal trafficking and toxicity of ENPs. Indirect contributions by work package Objective 9: Improve stakeholder understanding of ENP risks. Indirect contributions by work package Work package leader Dr. Alison Crossley (UOXF.DJ) Participants UOXF.DJ, NT, IHPP, CU, AXME, UGOT Objectives Manufacture, procurement and detailed characterisation of the two versions of each ENP (CeO2 , ZnO and Ag) provided by the NanoFATE industrial partners and selected as the focus for risk characterisation, fate, and effects experiments and risk assessment. This characterisation built on manufacturers' and existing data where possible and considered carefully the forms in which ENPs would be delivered (e.g. powders or suspensions, including issues of carriers and stabilisers). (Main participants: UOXF.DJ, NT, AXME) Production of ZnO ENPs tagged with rare earth elements in very low natural concentrations (e.g.Ytterbium or Luthetium) that match as closely as possible the ZnO nanoparticles common in commercial products. This is needed to allow tracking of the ZnO ENPs by ICP-based element analysis in the experiments to measure fate and effects. (For Ce and Ag based ENPs the natural background is so low they can be tracked by their core elements, while Zn has too high natural background for this to work if not tagged). An alternative approach, although significantly more expensive, is to synthesize the ZnO nanoparticles from a stable enriched isotope, e.g. 67 Zn or 70 Zn. (Main participant: IHPP) Characterisation of the ENPs used initially in the standard exposure medias of WP3. For ZnO this was done for both tagged and untagged ZnO to check how the behaviour of tagged particles matches that of untagged particles. (Main participants: IHPP, UOXF.DJ, UGOT) Validation of element-based analysis for gross tracking of ENPs. Method development and validation for the detailed tracking and localisation of ENPs in exposure and tissues relevant in WP2, WP3 and WP4 to refine methods for detecting particle in environmental media and in cells and tissue of exposed organisms. (Main participants: IHPP, UOXF.DJ, UGOT) Identification, quantification and characterisation of ENPs in samples provided from WPs 2, 3 and 4 to provide: (1) qualitative and quantitative data on environment-ENP interactions, and (2) the association of ENPs with both matrix and organism components in different environmental systems. (Main participants: IHPP, UOXF.DJ, UGOT, NERC, VUA, CU, UAVR) Note that the deliverables of WP1 were not public reports. However many lessons have been learnt that will aid future research at the level of characterisation. See our Advice Notes and also Newsletters for an indication of the work, and revisit the "legacy site" at the end of the project (Spring 2014) for a summary of lessons learnt. On 6th March 2014 our Training Course includes a module on: Making your own Gold NPs and characterising them and other NPs. Tasks 1.1) Manufacture, procurement and characterisation of ENPs. 1.2) Production and testing of the tagged versions of ENPs, and characterisation of ENPs in the environmental matrices of interest. 1.3) Tracking and characterisation of ENPs in the definitive fate and ecotoxicology experiments. Work package 2 ENP environmental behaviour and fate modelling Objective 2: Generate models for predicting the likely levels and states of ENPs in receiving waters and soils. Direct contributions by work package Objective 3: Analyse ENP interactions with environmental and biological entities using advanced microscope and physical analysis. Direct contributions by work package Objective 4: Study ENP fate and behaviour through wastewater treatment processes and in soils. Direct contributions by work package Objective 8: Develop risk assessment models(s) that integrate ENP fate, availability, accumulation and toxicity over the full post production life-cycle including provisions of data fro use in full life-cycle assessment. Indirect contributions by work package Objective 9: Improve stakeholder understanding of ENP risks. Indirect contributions by work package Work package leader Dr. Martin Hassellöv (UGOT) Participants NERC, UOXF.DJ, F+B, IHPP, AXME, UGOT Objectives Identify required input parameters to support analysis of scenarios of current and future ENP release developed in WP6 to parameterise initial worst case exposure assessments and derive predicted environmental concentrations for receiving aquatic and soil ecosystems. (Main participants: NERC, F+B, UGOT) Develop experimental methods to study ENP-environment interactions and their consequences for ENPs fate and behaviour through wastewater and sewage treatment processes including in receiving natural waters and sludge. (Main participants: UGOT, NERC, UOXF.DJ, IHPP) Apply refined experimental approach to investigate the effects of particle properties on ENP fate and behaviour through wastewater treatment processes and associated effluents. (Main participants: UGOT, NERC, UOXF.DJ, IHPP) Track effects of particle properties and environmental physicochemical properties on the rates of dissolution of ENPs during wastewater treatment and in a range of natural soils and waters. (Main participants: UGOT, UOXF.DJ, IHPP) Iteratively incorporate available information on ENP-environment interactions and their consequence for ENPs fate into the developed exposure models allowing for a refined PEC for waters and soils. (Main participant: NERC) Tasks 2.1) Development and refinement of Fate models. 2.2) Studies of ENP fate and behaviour through the wastewater treatment processes. 2.3) Building an understanding of how ENP properties and wastewater treatment processes and capability affect the final treatment success and fate of ENPs under current and predicted future loads. Work package 3 ENP ecotoxicology Objective 2: Generate models for predicting the likely levels and states of ENPs in receiving waters and soils. Direct contributions by work package Objective 5: Determine the chronic toxicity of ENPs of different properties, including co-exposures with other stressors. Direct contributions by work package Objective 6: Establish and model how environment physico-chemical properties in wastewater, natural waters and soil govern ENP parameters that each may ultimately affect bioavailability to organisms. Indirect contributions by work package Objective 7: Establish the mechanisms of uptake, internal trafficking and toxicity of ENPs. Indirect contributions by work package Objective 8: Develop risk assessment models(s) that integrate ENP fate, availability, accumulation and toxicity over the full post production life-cycle including provisions of data fro use in full life-cycle assessment. Indirect contributions by work package Objective 9: Improve stakeholder understanding of ENP risks. Indirect contributions by work package Work package leader Dr. Susana Loureiro (UAVR) Participants NERC, VUA, UOXF.DJ, UAVR, UNIPMN, IHPP, CU, AXME, UGOT Objectives Collate information on the suitability of existing ecotoxicological tests systems for ENP testing and subsequent optimisation of the NANOFATE biotest systems. (Main participants: NERC, VUA, UOXF.DJ, UAVR, UNIPMN, CU, UGOT) Ecotoxicological hazard characterisation of pure ENPs using the optimised media to establish chronic thresholds of concern (no observed effect concentrations, benchmark concentrations) for population relevant endpoints (e.g. reproduction) for selected aquatic and terrestrial test species. (Main participants: NERC, VUA, UOXF.DJ, UAVR, UNIPMN, CU, UGOT) Determination of the ecotoxicology of the selected ENP types in environmentally realistic situations for aquatic and terrestrial systems, including the assessment of interactive effects with physiochemical components of different exposure media and interactions with UV and organic chemicals as potential co-stressors. (Main participants: NERC, VUA, UOXF.DJ, UAVR, UNIPMN, UGOT) Identification of the major drivers for the ecotoxicological effects of ENPs, such as the contribution of the ENP itself and the metal ions they liberate, the coating of the ENPs after passage through STPs, the dependence on the exposed species and considered exposure route. Compilation of the experimental information and existing information from the literature and other R&D projects. Knowledge transfer to WP 4 (bioavailability studies) and WP 6 (risk assessment). (Main participants: NERC, VUA, UOXF.DJ, UAVR, UNIPMN, IHPP, CU, AXME, UGOT) Tasks 3.1) Establishment of exposure systems. 3.2) Ecotoxicological hazard characterisation of pure ENPs. 3.3) Advanced hazard characterisation of ENPs under environmentally realistic conditions, especially taking into account major confounding factors (natural media, UV and PAHs from combustion processes). 3.4) Synopsis: Identification of critical drivers for the ecotoxicology of ENPs. Work package 4 ENP bioavailability - relations between soil and water chemistry and particle properties Objective 3: Analyse ENP interactions with environmental and biological entities using advanced microscope and physical analysis. Indirect contributions by work package Objective 6: Establish and model how environment physico-chemical properties in wastewater, natural waters and soil govern ENP parameters that each may ultimately affect bioavailability to organisms. Direct contributions by work package Objective 8: Develop risk assessment models(s) that integrate ENP fate, availability, accumulation and toxicity over the full post production life-cycle including provisions of data fro use in full life-cycle assessment. Indirect contributions by work package Objective 9: Improve stakeholder understanding of ENP risks. Indirect contributions by work package Work package leader Dr. Kees van Gestel (VUA) Participants NERC, VUA, UOXF.DJ, UAVR, UNIPMN, IHPP, CU, UGOT Objectives Review and utilise existing data-holdings and where necessary primary research articles that contain information on ENP and media properties and their relationship with acute and chronic toxicity in aquatic and terrestrial species. (Main participants: VUA, UAVR, NERC) Establish and model how environment physicochemical properties govern ENP stability, soil--solution partitioning and transformation (e.g. dissolution) in natural waters and soils utilising time dependent kinetic modelling; provide chemical speciation modelling using established tools such as the Windermere Humic Acid Model and an adapted version of the Biotic Ligand Model including both particle dependent and dissolved metal ion toxicity. (Main participants: UGOT, NERC, VUA) Undertake specifically designed studies to investigate interactive effects among factors (ENP and media characteristics) that influence availability and uptake of ENPs in key species. (Main participants: NERC, UAVR, UOXF.DJ, UGOT, IHPP, DISAV) Develop kinetic models that describe environmental effects on particle dissolution rates and incorporate both free ion and particle associated toxicity to address the relationship between the relative contribution of ENP- and free metal ion-derived toxicity for exposed species. (Main participants: VUA, NERC) Tasks 4.1) Collation, data basing and mining of existing and newly available data. 4.2) Detailed experiments on how ENP and media properties interact to affect ENP presentation in environmental media and their availability for uptake. 4.3) Integrated assessment and modelling of ENP uptake and toxicity across soils and waters of varying chemical properties. Work package 5 ENP toxicokinetics and toxicodynamic Objective 5: Determine the chronic toxicity of ENPs of different properties, including co-exposures with other stressors. Indirect contributions by work package Objective 6: Establish and model how environment physico-chemical properties in wastewater, natural waters and soil govern ENP parameters that each may ultimately affect bioavailability to organisms. Direct contributions by work package Objective 7: Establish the mechanisms of uptake, internal trafficking and toxicity of ENPs. Direct contributions by work package Objective 9: Improve stakeholder understanding of ENP risks. Indirect contributions by work package Work package leader Dr. Francesco Dondero (UNIPMN) Participants NERC, VUA, UOXF.DJ, UAVR, UNIPMN, IHPP, CU, UGOT Objectives Identify the internal fate of trackable particles in exposed organisms by combining time series exposures with detailed tissue and cellular morphology using qualitative and quantitative particle detection methods. (Main participants: UNIPMN, CU, UOXF.DJ, IPHH) Investigate the link between internalisation and toxicological effects in time series exposures, and use model prediction of toxicokinetic parameters to study the internal tissue distribution and form of ENPs (using analytical electron microscopy) to attribute causal effects to intact ENPs and dissolved free metal ions. (Main participants: VUA, NERC, UGOT, CU, UOXF.DJ) Investigate the suitability of existing biomarkers for major modes of action known to be linked to ENP toxicity (e.g. reactive oxygen species (ROS) generation, lysosomal responses) for identification of ENP exposure and early-stage effects. (Main participants: UNIPMN, UWC/NERC, VUA, CU) Utilise a toxicogenomic approach to investigate novel modes of action associated with ENP exposure when compared to effects attributable to the free metal ions in samples from organisms exposed at ion levels close to concentrations known to cause life-history effects. (Main participants: UNIPMN, CU, VUA) Tasks 5.1) Modelling and measurement of ENP toxicodynamics and toxicokinetics. 5.2) Comparative mechanisms of action of dissolved, bulk, and nanoparticulate forms. Work package 6 Integrated environmental risk Objective 2: Generate models for predicting the likely levels and states of ENPs in receiving waters and soils. Direct contributions by work package Objective 6: Establish and model how environment physico-chemical properties in wastewater, natural waters and soil govern ENP parameters that each may ultimately affect bioavailability to organisms. Direct contributions by work package Objective 8: Develop risk assessment models(s) that integrate ENP fate, availability, accumulation and toxicity over the full post production life-cycle including provisions of data fro use in full life-cycle assessment. Direct contributions by work package Objective 9: Improve stakeholder understanding of ENP risks. Direct contributions by work package Work package leader Dr. Andrew Johnson (NERC) Participants NERC, VUA, UAVR, F+B, NT, UNIPMN, IHPP, CU, UGOT Objectives Conduct literature review and a stakeholder consultation to define current and potential future uses of ENPs and determine the driving parameters (including associated uncertainties) that define plausible future release scenarios. Collate consumption/discharge information and fate information (WP2) for use in exposure modelling. (Main participants: F+B, NERC, NT, AXME, SYMLOG) Conduct initial estimates of the concentrations of nanoparticles in different environmental compartments (air, sludge/soil, water and sediments) using the standard EU EUSES modelling approach based on the scenarios developed in Objective 1. (Main participants: NERC, F+B, UGOT) Generate robust Predicted No Effect Concentrations (PNECs) by integration of the experimental hazard data from WP3, the bioavailability data from WP4 and ecotoxicology data from the literature with the exposure assessments to develop a sound understanding of the environmental hazards of ENPs and how environmental media (water and soil) affect this. (Main participants: NERC, VUA, UGOT) Incorporate information on PNECs into multi-media (EUSES) and in GIS based models for soil (using ADMS) and receiving waters (using LF2000-WQX) to allow risk visualisation analyses for using generic and taxa specific PNEC and other endpoint data as appropriate. (Main participants: F+B, NERC) Undertake appraisal of the current and potential future environmental risks of CeO 2, ZnO and Ag ENPs, based on the work conducted in WPs 2, 3, 4, 5, including gap analysis relating to the uncertainties in hazard, fate and exposure assessment as well as the use and emission scenario definitions. (Main participants: NERC, VUA, F+B, UGOT, UAVR, CU) Develop appropriate project material to disseminate results and applied methodologies to relevant stakeholders, in order to improve understanding of ENP risks. (Main participants: NERC, VUA, F+B, UGOT, UAVR, CU, SYMLOG) Tasks 6.1) Literature review for scenario development. 6.2) Product information. 6.3) Exposure estimation. 6.4) Analysis of hazard and effect. 6.5) Gap analysis. 6.6) Development of dissemination material. Work package 7 Dissemination and training Objective 8: Develop risk assessment models(s) that integrate ENP fate, availability, accumulation and toxicity over the full post production life-cycle including provisions of data fro use in full life-cycle assessment. Direct contributions by work package Objective 9: Improve stakeholder understanding of ENP risks. Direct contributions by work package Work package leader Dr. Claire Mays (SYMLOG) Participants NERC, SYMLOG Objectives Develop dissemination plans, taking into account stakeholder needs. (Main participants: NERC, SYMLOG) Develop and maintain the electronic communication and dissemination platforms for the project during and beyond the lifetime of NanoFATE. (Main participants: NERC, SYMLOG) Develop internal training program. (Main participants: NERC, SYMLOG) Coordinate or perform their implementation. (Main participants: NERC, SYMLOG) Tasks 7.1) Dissemination and stakeholder interaction. 7.2) Newsletters; project electronic platform (data holding, intra and internet) including the NanoFATE legacy. 7.3) Training.
