Final Report Summary - FACEIT (Fast Advanced Cellular and Ecosystems Information Technologies)
The focus of FACEIT was pollution disaster management and in particular pollution disasters resulting from petroleum oil spills, either in marine or freshwater aquatic systems. The FACEIT project was carried out in the Thematic Sub-Priority Area 6.3 'Global change and ecosystems', with particular relevance to area III.3 'Risk assessment, management, conservation and rehabilitation options in relation to terrestrial and marine ecosystems'. The main position of the FACEIT consortium in the area of pollution disaster management was that of a strong biological actor.
Disaster management involves integrating different lines of information from and among different societal actors, such as pollution source, localisation, nature of chemicals involved, type of environment, responsible persons, or casualties. Essential elements of disaster management must include the rapid determination of the magnitude of the catastrophe, the protection of (aquatic) ecosystems, goods and persons from further damage and the suggestion of remediation measures which will restore the diversity and functioning of the affected ecosystem. The main focus of the FACEIT project was to provide a biomonitoring approach that can contribute to these three essential elements.
One of the key concepts of FACEIT was to use organisms, cell lines or biological molecules (DNA, RNA, proteins) to monitor pollution disasters and predict long-term effects and restoration. The importance of biomarkers has long been recognised and different endpoint markers are effectively used in monitoring programs. However, many biomarker tests were not sufficiently rapid or ethical and one of the goals of FACEIT was to redesign such tests by new combinations of methods or new concepts, such as by flow cytometric analyses of cell viability. In addition, FACEIT proposed to increasingly replace biomonitoring tests with unicellular organisms or cell-lines, which have the primary advantage of being much more rapid than tests with whole complex organisms and potentially reduce measurement costs. However, by proposing this, FACEIT had to demonstrate that responses measured in unicellular organisms or cell lines can be extrapolated and validated to effects on whole organisms and ecosystems, which is one of the most important research objectives of the program. Essentially, no tests existed at the starting date which monitor and predict medium- and long-term consequences of a pollution disaster for ecosystems. FACEIT, therefore, proposed a few key areas for new method developments. These were: biological diversity, multifunctionality or self-regeneration capacity, specifically focused on petroleum hydrocarbon pollution and its bioavailability, microbial communities, and their biodegradation potential.
The consortium was strongly convinced that biological monitoring tools for affected ecosystems are an essential element of disaster management, and should be used by other actors in the landscape of disaster management to aid in decision making and in developing remediation strategies. They realised therefore, that the tools and methods developed within the realm of FACEIT can only make the transition to management and application if they were sufficiently supported by research findings, validation and training. In order to promote the use of biomonitoring tools, which could lead to their testing and validation by third persons, FACEIT attempted to actively inform different communities of actors in the disaster management 'landscape' or via web-based resources. Apart from various brochures, press releases, smaller meetings, this has culminated in one 'hands-on' training course in July 2008, a final two-day symposium in January 2009 and a new handbook on microbiology of hydrocarbons, oils and lipids appeared in summer 2009 with Springer Publishers.
One of the main technological developments in FACEIT was the application of bacterial reporter assays. Such assays consist of relatively simple incubations in aqueous media with genetically modified bacteria, engineered to produce bioluminescence or fluorescence in contact to a set of target chemicals. A large number of reporter strains were developed targeting key components of oil (e.g. alkanes, BTEX, PAHs). Overall, this work package has been very successful and most of its goals could be realised. A concise set of bioreporter strains with the same chassis and reporter output for use in assays to detect key oil-borne pollutants was developed. Different assay formats were designed, extensively calibrated and tested by various users, which makes us confident that we have realised a very simple and rapid set of assays to quantitatively analyse oil pollution in water with bacterial bioreporters. The assays were also validated on a real-life scenario of an open sea spill and were robust to be carried on board and perform analysis in the first two hours after a spill. We thus feel very confident that we have produced a potentially very useful tool for spill analysis that can find its place among a biotool set. The concept of bioreporter assays was further taken up in instrumentation and product development. A semicontinuous standalone bacterial bioreporter system for on-line analysis of general inducible stress was instrumentalised. A functional eight chamber system was developed based upon the so-called ROTAS system. Within this system freeze-dried bioreporter strains were capable of being rehydrated and functioning effectively in response to contaminated samples. A limitation of the work was the propensity of the peizoactuated microfluidic pumps to block and the need for upfront filter development, however, a few effective prototypes were generated.
FACEIT has added a number of useful tools and concepts for disaster management strategies. However, not one tool will hold all, and regulatory agencies or policy and decision makers often require a single integrated value to assess the risks of contamination. FACEIT hereto favours integrative approaches, which would lead to colour scales or a single numbering system for quality assessment.
Disaster management involves integrating different lines of information from and among different societal actors, such as pollution source, localisation, nature of chemicals involved, type of environment, responsible persons, or casualties. Essential elements of disaster management must include the rapid determination of the magnitude of the catastrophe, the protection of (aquatic) ecosystems, goods and persons from further damage and the suggestion of remediation measures which will restore the diversity and functioning of the affected ecosystem. The main focus of the FACEIT project was to provide a biomonitoring approach that can contribute to these three essential elements.
One of the key concepts of FACEIT was to use organisms, cell lines or biological molecules (DNA, RNA, proteins) to monitor pollution disasters and predict long-term effects and restoration. The importance of biomarkers has long been recognised and different endpoint markers are effectively used in monitoring programs. However, many biomarker tests were not sufficiently rapid or ethical and one of the goals of FACEIT was to redesign such tests by new combinations of methods or new concepts, such as by flow cytometric analyses of cell viability. In addition, FACEIT proposed to increasingly replace biomonitoring tests with unicellular organisms or cell-lines, which have the primary advantage of being much more rapid than tests with whole complex organisms and potentially reduce measurement costs. However, by proposing this, FACEIT had to demonstrate that responses measured in unicellular organisms or cell lines can be extrapolated and validated to effects on whole organisms and ecosystems, which is one of the most important research objectives of the program. Essentially, no tests existed at the starting date which monitor and predict medium- and long-term consequences of a pollution disaster for ecosystems. FACEIT, therefore, proposed a few key areas for new method developments. These were: biological diversity, multifunctionality or self-regeneration capacity, specifically focused on petroleum hydrocarbon pollution and its bioavailability, microbial communities, and their biodegradation potential.
The consortium was strongly convinced that biological monitoring tools for affected ecosystems are an essential element of disaster management, and should be used by other actors in the landscape of disaster management to aid in decision making and in developing remediation strategies. They realised therefore, that the tools and methods developed within the realm of FACEIT can only make the transition to management and application if they were sufficiently supported by research findings, validation and training. In order to promote the use of biomonitoring tools, which could lead to their testing and validation by third persons, FACEIT attempted to actively inform different communities of actors in the disaster management 'landscape' or via web-based resources. Apart from various brochures, press releases, smaller meetings, this has culminated in one 'hands-on' training course in July 2008, a final two-day symposium in January 2009 and a new handbook on microbiology of hydrocarbons, oils and lipids appeared in summer 2009 with Springer Publishers.
One of the main technological developments in FACEIT was the application of bacterial reporter assays. Such assays consist of relatively simple incubations in aqueous media with genetically modified bacteria, engineered to produce bioluminescence or fluorescence in contact to a set of target chemicals. A large number of reporter strains were developed targeting key components of oil (e.g. alkanes, BTEX, PAHs). Overall, this work package has been very successful and most of its goals could be realised. A concise set of bioreporter strains with the same chassis and reporter output for use in assays to detect key oil-borne pollutants was developed. Different assay formats were designed, extensively calibrated and tested by various users, which makes us confident that we have realised a very simple and rapid set of assays to quantitatively analyse oil pollution in water with bacterial bioreporters. The assays were also validated on a real-life scenario of an open sea spill and were robust to be carried on board and perform analysis in the first two hours after a spill. We thus feel very confident that we have produced a potentially very useful tool for spill analysis that can find its place among a biotool set. The concept of bioreporter assays was further taken up in instrumentation and product development. A semicontinuous standalone bacterial bioreporter system for on-line analysis of general inducible stress was instrumentalised. A functional eight chamber system was developed based upon the so-called ROTAS system. Within this system freeze-dried bioreporter strains were capable of being rehydrated and functioning effectively in response to contaminated samples. A limitation of the work was the propensity of the peizoactuated microfluidic pumps to block and the need for upfront filter development, however, a few effective prototypes were generated.
FACEIT has added a number of useful tools and concepts for disaster management strategies. However, not one tool will hold all, and regulatory agencies or policy and decision makers often require a single integrated value to assess the risks of contamination. FACEIT hereto favours integrative approaches, which would lead to colour scales or a single numbering system for quality assessment.
