Biosensors & Bioelectronics 14 (1999) 541 544 www.elsevier.

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BIOSET: Biosensors for Environmental Technology EU Workshop Biosensors for Environmental Monitoring: Technology Evaluation Held in Kinsale, Ireland, 1215 May 1998.
C. OSullivan a, S.J. Alcock b
a

Uni6ersity College Cork, Cork, Ireland b Craneld Uni6ersity, Craneld, UK

Received 3 January 1999; received in revised form 17 May 1999

Biosensors combine the power of microelectronics with the selectivity and sensitivity of biological components such as whole cells, organelles or biomolecules e.g. antibodies, receptors, enzymes and nucleic acids. Biosensors offer unique advantages for environmental monitoring because of their specicity, fast response times, low cost, ease of use and continuous real-time signal. Biosensors can detect biological effects such as genotoxicity, immunotoxicity and endocrine responses and can be used to detect individual substances or groups of substances in the environment, such as industrial emissions and can also be used for control of domestic-waste water treatment and ensuring the success of sanitary measures. The universal applicability of biosensors places them clearly at the core of any programme to address future technology for environmental monitoring in real matrices. In many monitoring situations biosensors can be expected to be the best available technology not entailing excessive costs. BIOSET, the EU Concerted Action on biosensors for environmental monitoring/environmental technology, has the role of guiding technological developments in the eld of biosensors for environmental monitoring. The aim is to enhance the development of biosensors for practical applications in monitoring environmental pollutants in water, air, soil and waste. A forum is provided for exchange of information, involving universities, research centres, manufacturers and end users in order to facilitate the transfer of knowledge to industry and others and to identify gaps in knowledge and the respective research and development projects to address them. BIOSET will enhance understanding of new analytical demands, including new parameters to describe

environmental phenomena and effects, and the potential of biosensors to satisfy these demands. Priority is being given to development of existing biosensors for routine application in environmental monitoring, measurement of new parameters of relevance to the environment, and the development of integrated sensor systems capable of measuring several parameters simultaneously under real operational conditions. The rst full Concerted Action Workshop entitled Biosensors for Environmental Monitoring: Technical Evaluation, was held in Kinsale, Ireland, 1215 May 1998. The Workshop was attended by participants from the UK (11), Ireland (18), Germany (11), Spain (5), Italy (3), Portugal (2), France (2) and Belgium, Sweden, Norway, Greece, Austria and Israel (1 each), a total of 58 delegates. The heart of the scientic content was based on 26 high quality presentations, ve discussions and 20 posters. The workshop was opened by Prof. Guilbault (University College Cork) and Dr Busing (European Commission), who warmly welcomed all the delegates before Prof. Turner and Dr Alcock (Craneld University) introduced BIOSET and highlighted the objectives, current and future activities and overall aim of the Concerted Action. The project involves four major tasks: Involvement of industrial producers and users in addressing problems of mass production and of performance in real conditions, in order to bring to fruition the useful application of existing biosensors. Enhancement of understanding of new analytical demands, including new parameters to describe environmental phenomena and effects, and the potential of biosensors to satisfy these demands.

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Denition of specic environmental tasks as targets for biosensor development. Identication of paths to resolve the most effective biosensors for integrated management of industrial waste (including air contaminants) and for use in wastewater management in order to promote a clean environment. Dr Alcock reported on the objectives that had been achieved, the scientic and technical progress and the role of publication and technology transfer. Prof. Fiksdal (Norwegian University of Science and Technology) reported on the rst BIOSET focus workshop, which was held in Trondheim, Norway on 3 4 April 1998 and Prof. Hansen (TU Berlin) summarised the main results of the biosensor eld experiment that took place on 1619th September, 1997 in a municipal monitoring station in the State of Brandenburg in Germany. Mr. Nowak (DNU Nowak Umweltanalysen, Berlin) went on to give a demonstration of the CDROM results of this biosensor eld experiment. Profs. Guilbault (University College Cork) and Mascini (University of Firenze) gave an overview of the limitations in the application of existing biosensors, receptors and transducers to monitor pollutants in environmental matrices which was followed by an in depth discussion on the topic. This discussion highlighted the limitations of the application of existing biosensors as well as bringing to attention the fact that many existing biosensors are useful but need to be utilised in a more focused manner, i.e. to compliment existing technologies and to identify user requirements and tailor biosensor design to meet these needs. Prof. Dominguez (University of Alcala, Madrid) began the in-depth discussion by stressing that biosensors can introduce new information which is not available by other means. Dr Busing, who added that the aim of biosensor development should be to see what additional information can be gleaned with biosensors rather than seeking to replace existing methods, supported this opinion. Dr Etienne (CNRS, Paris) continued by giving an example of such a use, citing the ability of biosensors to be utilised to determine bioeffects as well as analyse presence/concentration. This viewpoint was also upheld by Dr Forrow (Environment Agency for England and Wales), who quoted the CARACAS report that highlighted the need for regulators to have available effects-based methods, a requirement that could be met by biosensors. Prof. Hansen proceeded with the discussion by emphasising, for example, the usefulness of an immunotoxicity index and Prof. Mascini talked about the application of sensors to eld use and as fast alarm systems. Prof. Guilbault continued with this theme by commenting that the B.O.D. sensor had been a success and there was a real potential application for sensors for xenobiotics and nitrates, nitrites and phenols amongst others but went on to say

that the key to successful commercialisation was to have a high volume, low cost market more applicable to the clinical market than the environmental sector at the moment. Another slant presented in the discussion was the necessity to focus on the nal use of the biosensor. Dr Busing stressed the need to have clearly dened objec tives and to be specic in these aims (e.g. to develop a biosensor specically for process control for a sewage plant and not broadly for the environment). Dr Steinwand (Bodenseewerk Perkin Elmer GmbH) picked up the discussion, commenting that the nal user of the biosensor has a heavy inuence on how the product is designed. This was further elaborated on by Prof. Coulet (University of Lyon) who stressed that the user needs results that he can relate to. The discussion was concluded with the question posed as to why biosensors have focused on water analysis and have not yet been established for air/soil monitoring. This question was replied to by Prof. Turner who described the difculties of getting biological systems to withstand exposure in air, but gave some examples of success with biosensors for both gas and soil monitoring. Dr Garnham (BNFL, Preston) outlined the future challenges in environmental monitoring and commented that the future strategy of BNFL in the reprocessing of nuclear fuels was the use of on-line, novel and cheap sensors to monitor efuent and other processes in plants. Dr Forrow presented information on the UK Demonstration Programme on Direct Toxicity Assessment. In the UK limits are set on discharge controls to control the release of single substances to fall into line with EU guidelines. Currently, there are 100 000 chemicals on the market with this number growing by 200 annually. In addition to these known chemicals, unknown substances exist and furthermore there can be interaction between substances. In some cases, it is possible to have more than 20 000 substances in a single efuent. As every substance cannot be monitored, the Environment Agency has derived protocols for toxicitybased control of efuents. Rapid tests are thus required for cost-effective monitoring and it is in this area that biosensors could contribute, either as broad-spectrum toxicity biosensors or more specic diagnostic biosensors. The coupling of these two types of biosensors could lead to tracing of toxicity sources up waste streams and the potential to track and identify responsible chemicals. Dr Forrow concluded by re-emphasising that efuent control is a promising area for biosensors and that groups should be encouraged to interact with end-users. Prof. Dominguez gave a highly insightful talk on the most promising technologies to enhance sensors. She began by highlighting the fact that the goal of environ-

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mental analysis is sustainable development, providing more from less in order to ensure quality of life. She proceeded by stressing the importance of focused data analysis to solve the problem in question. Old characterised systems can be used in new applications (e.g. glucose, lactate, gas-phase detection of phenol) and the use of novel orientated immobilisation techniques (gene fusion, site-directed mutagenesis, self-assembled monolayers and Langmuir Blodgett lms) can make these systems more effective. A new application of biosensors is toxicity screening; biosensors are particularly suitable, being both cost-effective and adaptable to the mode of continuous operation an example of an area for potential biosensor application is the continuous analysis of water for Cryptosporidium, which currently claims the lives of 11 000 children every day. Prof. Dominguez proceeded by saying that she felt the area of DNA sensing is promising and perhaps superior to that of immunosensors, which are wholly dependent on antibody production, screening and purication, before concluding her talk by examining the pros and cons of the available transducers. Dr Busing added that in the future very large projects may be more suitable, and cited the example of the yeast genome project that involves about 100 laboratories which is highly benecial to all participants by sharing data and ideas. Prof. Turner picked up on this theme at a later stage in the workshop, focusing on strategies to enhance the performance of biosensors. He began by commenting that biosensors can operate selectively in real matrices but that there are major gaps in the engineering of complete systems, particularly in the areas of separation science, uidics, measurement, calibration, maintenance and data processing. A principal point Prof. Turner made was that there are limited further opportunities for single analyte biosensors and the most promising route to follow is that of the development of multichannel biosensors, as the market demands adaptable multianalyte devices. This area, however, requires some further advancement, particularly in the area of array fabrication, miniaturisation, microuidics, regeneration, stability, adaptability and chemometrics. Another area for exploitation is the combination of the inherent advantages of biological components with the reproducibility of synthetic chemical biomimics, the latter being more stable, cheaper and more amenable to deposition in arrays. Industries that could be targeted include the water, leather, paper and textile industries and these could be used to illustrate the usefulness of biosensors. Industry and regulatory bodies need to accept biosensors in terms of robustness and convenience and thus, if biological components are to be utilised, advances in strategies to enhance stability are needed. In conclusion, Prof. Turner highlighted the major priorities for further progression as: (i) identication of stable receptors for key analyses/effects; (ii)

sensing systems with an ability to yield required analytical performance in real matrices; (iii) fabrication and operation of microchannel arrays; (iv) engineering of integrated systems for specic applications and nally (v) a true appreciation of the market and legislative requirements. Prof. Cammann (ICB, Munster) followed this up by presenting results of a parallel Concerted Action on Biosensor Stability (funded under the EU Industrial and Materials Technologies Programme), providing useful information to aid in the development of effective biosensors for environmental monitoring. Dr Alcock gave an Internet demonstration on biosensors for environmental monitoring and Ms. Barzen (University of Tubingen) demonstrated a biosensor device. Thirteen groups funded under the EU Environment and Climate programme gave high quality scientic reports on the status of their collaborative projects. Mr. Nowak led a discussion on the involvement of SMEs in BIOSET and asked why so few companies attend Biosensors for Environmental Monitoring workshops. Mr. ODonovan (University College Cork) of Audit Diagnostics, a locally based SME, replied that SMEs require some form of funding and stressed that small companies are full of ideas and enthusiasm but do not have access to extra funds nor do they have their own R&D facilities. Dr Schneider of Sension GmbH, Augsburg, added that it is very important to have innovative and energetic partners and that working with SMEs can be very fruitful in comparison to large companies, who may only be interested if there is a nished product that is ready to sell. Prof. Turner said that one-to-one personal contacts can lead to real involvement e.g. in EU projects and Dr Busing sup ported this, commenting that the CRAFT network has national focal points for SMEs and that focus meetings with these SMEs may be arranged. Dr Pfeiffer (BST, Berlin) concluded this discussion by pointing out the fact that SMEs need real support, both nancial and technical, at the prototype stage, adding that a prototype can be extremely expensive and mass production of the device cannot be assessed without knowing exactly the market capacity. A poster session was held with representation from most participating groups. A selection committee (Prof. Dominguez, Prof. Hansen and Dr Barcelo (CID-CSIC, Barcelona) shortlisted three posters: 1. Integrated Mach-Zehnder sensors based on arrow structures as optimised transducers for biosensing technology, Lechuga, L.M., Prieto, F., Calle, A. and Dominguez, C., IMM-CMN, Spain. 2. Competitive assay for the herbicide 2,4dichlorophenoxyacetic acid using imprinted polymers and electrochemical detection, Kroger, S., Turner, APF., Craneld University, England; Mosbach, K. and Haupt K., Lund University, Sweden.

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3. Assay for Okadaic Acid, Kreuzer, M.P., OSullivan, C.K. and Guilbault G.G., University College Cork, Ireland. The BIOSET Steering Committee awarded a prize for the best poster to Mr. Kreuzer (Cork). Dr Brecht (University of Tubingen) gave an overview of integrated sensor systems for simultaneous measurement of multiple parameters. The logic behind the use of multiparameter systems is for use in legislative problems, the making of decisions, as alarms, for direct feedback on process control, to improve resolution and overall to give a better understanding of a process or a situation. The rationale for incorporating a sensor system is its cost-effectiveness and speed and a multiparameter sensor would provide much more useful information. Examples include the coulometric detection of heavy metals, the hyphenation of separation steps and on-line bioassays, miniaturised electrophoretic systems and multisensor systems with discrete sensing elements. An example of a commercially successful integrated sensor system for multiple parameter analysis is the i-STAT, which is a small, point-of-care analysis system based on multiparameter sensors, (Na + , K + , Cl , Ca2 + , pH, pCO2, pO2, glucose, urea). It is a reagentless, disposable system that can work either as a hand-held device or linked up to other equipment. No additional research was involved in its inception, simply a development of existing technologies. An example of multi-parameter detection with afnity-based systems is the Ekins Microspot test by Roche (formerly Boehringer). In summary, the technology for multisensor systems is available, and the fruition of such a commercial device depends on the sensor concept and on the need for such a system. Finally, Dr Brecht highlighted some niche areas for biosensors such as waste deposit sites (monitoring of run-off); on site analysis (site clean-up) and analysis of drinking water. The nal of eight sessions was chaired by Dr J. Busing of the European Commission and focused on EU Framework Programme 5, future priorities and conclusions of the workshop. Dr Busing began by saying that there will be no specic funding for biosensors in Framework 5 and that there will be fewer programmes than in Framework 4, with four or ve themes being highlighted and the emphasis being very much problem-orientated. A clustering of projects will be favoured with basic research receiving funding where appropriate. In principle, it will be more difcult for biosensor experts to come together in meetings in the

future, but he commented that there will be useful Concerted Actions. In preparing project proposals, the projects should not be too ambitious and should start at a point where the chance of success is high. SMEs will still be important but Framework 5 will be more open to industry in general. It is important to maintain contact with the users in order to identify exact problems. Moreover, it will be useful to take an already developed idea and to create common testing platforms where technologies can be tested. The most advisable path to take is to develop an overall picture, predene the cluster groups and then invite groups to undertake specic components. All Concerted Actions inform the EU of their scientic information and in this way there is an easily accessible database where scientists can nd information. BIOSET is the most effective and efcient Concerted Action with an exchange system set-up as well as effective dissemination of results etc. The tools that have been set up by the Concerted Actions should be used and exchange and dialogue emphasised. The BIOSET group should show visibility, and press releases are very important to the EU. In closing this talk, Dr Busing stressed that BIOSET should, further more, be known as a centre of excellence and this is the image that should be developed. As a conclusion to the workshop, Dr Busing led a discussion on what had been achieved and what information had been gleaned. The message of the Concerted Action should be taken and disseminated to promote awareness of available information and platforms for exchange and dialogue and all co-ordinators of EU funded projects should undertake to inform their partners of all signicant points made. The BIOSET team is orientated to solving problems and, although there is a huge task ahead, with the mechanisms created by the Concerted Action ideas may be submitted and various groups informed and brought together in an effort to have the rst commercial environmental biosensors as a reliable agship. This highlights the usefulness of the Concerted Action.

Acknowledgements The organisers would like to thank the European Commission, University College Cork and the Royal Society of Chemistry for nancial support. For further information about BIOSET please contact s.alcock@craneld.ac.uk