Team:Utrecht/Human Practices

Synthetic biology is a rapidly evolving field, with a bright future perspective. Synthetic biology-based systems are sustainable, innovative and have many possibilities. While developing systems it is important to look for the most suitable, safe and widely accepted application. To determine what problem would be most relevant and best suitable for our system, it was essential to acquire information of both experts and stakeholders. This shaped the focus of our social relevance, which started as a detection method for botulism and shifted towards a system aiding detection of oestrogens and oestrogen-like compounds. Furthermore, the extensive feedback aided the design of a device that is convenient to use, low-cost, safe and meets all other stakeholder requirements. To give an impression of the evolution of our project, the project timeline is illustrated below. A summary of the most influential insights is provided in the last section, underneath the infographic.

Pim Klaassen, Niek Savelkoul, Jaco Westra and Korienke Smit, RIVM


About RIVM

The RIVM is the Dutch National Institute for Public Health and the Environment. They conduct research and provide advice to government authorities to improve policy relating to environmental and public health and safety. One of their core focusses is on policy surrounding safe use of GMO.

  • The development of a general lab tool is less interesting and powerful than an applied solution.
  • Contacting relevant organizations and businesses to figure out their needs is a good way to determine if problems in their field could benefit from our project.
  • Fields of interest could be food safety, environmental pollution, water quality, and chemical compounds.
  • It is likely unfeasible to engineer our GMO biosensor in such a way that introduction in the environment could legally be allowed.
  • Changed the aim of our project from a general lab tool to a specified sensor-solution.
  • Established contact with relevant organizations to catalogue interesting applications of our biosensor platform.

Jos van der Vosse, TNO


About Jos van der Vosse

Jos van der Vosse is a senior scientist, currently working for the Netherlands Organisation for Applied Scientific Research (TNO), where he develops detection systems to improve food quality and to ensure microbiological quality and safety.

  • Measuring specific contaminant concentration is crucial for application in food industry.
  • When using our biosensor for detecting contaminants, such as hormones and medicines in surface water, a less specific concentration might be sufficient.
  • A method using quantitative measurement is preferred. This technique does not require the emitted light to be visible for the eye.
  • Our method seems to be preferable above other methods, because in the future we might also be able to locate the source of the contamination.
  • Change in social relevance to surface water contamination
  • End product must quantitatively measure light
  • Designing an end product able to safely detect contamination source has high potential
Full Summary

Onno Epema, Rijkswaterstaat


About Onno Epema

Onno Epema is Head of Laboratory of Inorganic analysis at Rijkswaterstaat. One of the tasks of Rijkswaterstaat is the control of surface water quality. Its multiple measuring stations use both chemical, as well as biological systems to examine water quality.

  • The presence of pharmaceutical waste products in water is a relevant issue.
  • Toxicity of compounds in water is currently measured using bioassays. These assays include daphnia, fish or bacteria.
  • Additional chemical analysis to bioassay analysis is required to specifically identify, and quantify the presence of the toxic compound.
  • Direct identification of compound class (e.g. functional groups, medicinal group, hormones etc.) would be a leap forward in identification efficiency in chemical analysis. Information obtained during chemical analysis is often complex, as there are tens of thousands of potential toxic compounds in the water.
  • DeTaXion could potentially replace the microtox assay, providing a method which can both detect the presence, and concentration of toxic pharmaceutical waste products.
  • Confirmation of the importance of our biosensors to detect multiple concentrations.
  • The end product should be able to detect compounds classes instead of one specific compound. This is possible with the expression of multiple modified Tar receptors, each with a specific ligand, such as hormones.
Full Summary

Roberd Boer, Tauw


About Roberd Boer

Roberd Boer is currently working at Tauw Group and focuses on testing wastewater for issuing permits for discharge into the sewage system. The Tauw Group is an European consultancy and engineer agency, focusing on environment and sustainable development.

  • Taking a correct water sample is harder than it looks. The Tauw Group recommended the Nen 6600-1 method, a strict protocol aiming to sample the water correctly.
  • Waste product detection in water is based on the activities of the company. For pharmaceutical companies, the raw materials to produce the drugs are tested, not the drug itself as this is carefully filtered out of the water. Pharmaceutical water pollution is mostly caused by hospital waste water, which commonly contain: solvents, Iodide, Bromide, volatile aromatic hydrocarbons and heavy metals.
  • Currently, most assays are based on mass spectrometry, specified on the contaminant. These techniques show results within 24 hours.
  • It would be a great advantage that some biosensors are able to detect various compounds in wastewater. This way, the efficiency of the water purification and the concentration of contaminants can be measured at the source.
  • Shift the focus of measuring the presence of drugs to the presence of drugs residues.
  • Integrate the matter of difficulties of water sampling into the end product.
  • It will be an advantage if our end product can measure multiple compounds or groups of compounds at the same time.
Full Summary

Gert-Jan Euverink, Wetsus


About Gert-Jan Euverink

Prof. dr. Gert-Jan Euverink is Professor of the Faculty of Science and Engineering from the University of Groningen, where he was a supervisor of the winning iGEM team Groningen several years ago. Furthermore, he is deputy scientific director of Wetsus, Centre for sustainable Water Technology in Leeuwarden.

  • On existing biosensors:
    • A biosensor used currently at Wetsus is the lux system. This system utilizes an E.coli-based luciferase assay, which will emit light when a carcinogen is present.
    • Other biosensors are fish and daphnia, which rely on the behaviour of these organisms, resulting in a lot of false positives.
  • On our project:
    • Difficulties: docking molecules in an active site is rather difficult, multiple techniques can be used to tackle the problems
    • Random mutagenesis
    • These techniques require a lot of time or luck.
    • Dictyostelium bacteria have a stronger chemotaxis-mediated swarming effect than E. coli.
  • On the safety of our product:
    • When using this kind of biosensor, materials can be easily disinfected. Cleaning-up the water after the measurements might be harder, because you have to be certain all bacteria are eliminated.
  • Acquiring info about Dictyostelium bacteria to make sure the organism with the highest potential is used in our system.
  • Focussing on effective ways of sterilization and cleaning of the water used by the biosensor.

Martin van den Berg


About Martin van den Berg

Prof. Dr. Martin van den Berg is Professor in Toxicology, deputy director of the Institute of Risk Assessment Sciences(formerly RITOX) of the University of Utrecht and head of the Toxicology and Pharmacology Division of IRAS.

  • Waste water levels of selective serotonin reuptake inhibitors (SSRI) have increased to levels that influences the behaviour of fish and birds. SSRIs are pharmacologically active in vertebrates, as they use similar serotonin systems. Thus, SSRIs potentially affects other smaller animals in lower concentrations.
  • WSSRIs are soluble in water and will not easily evaporate. Therefore, we should study the effect of the SSRIs on the aquatic environment first.
  • The receptor might not be as selective in ditch water as in filtered water.
  • Methodological analyses is needed to investigate the effect of waste water on false positives/negatives.
  • E. coli is common in the human body and can survive and multiply easily in various circumstances. When a strain gets pathogenic by accident and released in the environment, it could have severe outcomes for society.
  • Find more on the social relevance of SSRIs and their impact.
  • We will apply a purification step before measuring to lose lipids and other substances which disturb the outcome.
  • In the end product, there have to be measures to make sure the bacteria can only survive in the device to prevent bacterial leak into the environment.
Full Summary

Marijan Uytewaal-Aarts and Marlies Verhoeven, Hoogheemraadschap de Stichtse Rijnlanden (HRSR)


About Marlies Verhoeven and Marijan Uytewaal-Aarts

Marlies Verhoeven works in sewage treatment for HRSR and is process technologist. Marijan Uytewaal-Aarts used to work for KWR in analysis of microbiology and toxicology. Now she works for HRSR as advisor of monitoring chemical water analysis.

  • Drug use and thereby the concentration of drug residues in surface water has increased steadily over past years and will continue to grow in the future.
  • Water quality is also checked at treatment plants to measure the efficiency of water purification.
  • Dutch surface water is a source for drinking water. Therefore, compounds in the surface water end up in the drinking water, posing a threat for public health.
  • There is a purification obligation in horticulture.
  • There are no set rules on the concentration of drug residues. Also, determining the effects of cocktails or low concentrations of drug residues is very difficult.
  • Our biosensor potentially replaces animal behavioural toxicity test, reducing the use of experimental animals.
  • Constant feedback and reflections on our project is needed, due to the use of GMO’s.
  • On potential stakeholders:
    • Companies producing or using pharmaceuticals and other chemical compounds are also stakeholders, since they have a purification obligation.
    • Water treatment plants also require concentration measuring methods.
  • On our biosensor:
    • Water samples should be filtered before applying it to our biosensor. As well, we need to make sure our biosensor survives in surface water and not only water used in laboratories (Milli Q water).
    • Consider techniques needed to test for E. coli presence in case some kind of leak in the system occurs.
Full Summary

Michael Bentvelsen, Dutch Water Authorities


About Michael Bentvelsen

Michael Bentvelsen is a policy advisor for the Dutch Water Authorities (Unie van Waterschappen), that represents all Water Authorities in the Netherlands. Water authorities are responsible for both water quality and water purification.

  • There is a variety of filter methods that can be used for water purification needed for water analysis.
  • For water purification, BDS uses a method with one purification step, in stead of the chemical purification methods, which needs two more expensive steps.
  • Although the government seems a logical stakeholder of biosensors, in practice the producers of the specific compounds, like pharmaceutical companies, are the major clients of BDS biosensors.
  • Biosensors have great advantages compared to chemical analysis methods. They are cheaper, faster and often measure a broader range of compounds.
  • End product integration:
    • Experiments have to be conducted to determine what filter method will be sufficient for DeTaXion. The needed filter methods are more likely to be similar to the methods used by BDS than by chemical analysis, because the systems of DeTaXion and BDS biosensors are more alike.
    • So far, we have set our minds on producing a kit, enabling buyers to do the experiments by themselves. However, end product design will be easier and the results will be more reliable when the measurements are done at the lab. We will have to reconsider our choices based on these newly acquired advantages and disadvantages.

Peter Behnisch, Biodetection Systems


About Peter Behnisch

Peter Behnisch is Chief Commercial Officer of the Dutch company BioDetection Systems b.v. (BDS). This company is specialized in bio-based screening technologies for safety, quality and bioactivity assessment.

  • Chemical analysis most often used, but very costly and often irrelevant results.
  • Water authorities are already familiar with bioassays. SIMONI and Daphnia-dependent assays are currently used methods. Since these techniques mostly rely on organism behaviour, they are not always reliable and measurements can be disturbed due to polluted water.
  • There are no national or European policies yet, although water authorities in the Netherlands are stimulated to approaching the problem.
  • There is a purification obligation in horticulture.
  • Ensuring the low-costs and high speed of DeTaXion, will make it a very beneficial and applicable system.
  • Confirmation that DeTaXion could ease chemical analysis and could be used as a precheck for chemical analysis.
  • Potential applications of DeTaXion would involve local measurements, such as:
    • Method for the detection of the health of the neighborhood
    • A quality check of water purification at the source.
    • Efficiency of pharmaceutical uptake of patients
    • Method to confirm predictions and national measurements
Full Summary

Marlies Kampschreur and Maarten Nederlof, Waterschap Aa and Maas


About Marlies Kampschreur and Maarten Nederlof.

Maarten Nederlof is involved in water purification by sewage plants and Marlies Kampschreur is responsible for the innovation at Aa and Maas. Water Authority Aa and Maas is responsible for water quality and water purification for their district.

  • Identifying the impact and setting a concentration limit of compounds or mixtures of compounds is rather difficult.
  • Pilots on techniques with a higher yield of water purification are being done. It is important that these techniques are as cheap as possible.
  • Germany and Switzerland already have such high yield techniques. In the Netherlands a pilot is held.
  • There is an increase in bioassays being developed and used. Water authorities are testing whether these can be meaningful to determining the effect of compound mixtures in water on organisms, rather than the existing compounds. SIMONI counts as golden standard. a follow up on the compounds with an effect is still needed.
  • The disposal of waste from illegal drug laboratories led to shutting down sewage treatment companies.
  • Considering the designing of an end product that can track down illegal drug laboratories.
Full Summary

Integration of human practices into our project

The input of the stakeholders, experts and public considerably influenced our project, altering the application of our sensor from a biosensor introduced in the environment to a handheld-device. Safety, low-costs and rapidly available quantified data were leading values in both project and product design processes. All requirements and difficulties emanating from the interactions were considered and integrated into the project, which is summarized below.

Finding a suitable application

We commenced with a method detecting botulism toxin in surface water, a serious issue for both wild animals and humans. In this case, quick demonstration of botulism presence and finding its source is important. However, upon discussing the topic with experts of the National Institute for Public Health and the Environment (RIVM), we decided that introduction of GMO into the environment would not be feasible. Therefore, we shifted our attention to the application of the sensor as a lab tool to measure the presence of a certain compound.

Since our system was not specifically aimed at botulism toxins, the decision was made to detect toxins in food. However, after a literature search we concluded that the size of toxins would be too large to be able to be detected by our system. This lead us to focus on the detection of small molecules in food. Jos van der Vosse from the Netherlands Organisation for applied scientific research (TNO) advised us to focus on the detection of small molecules in water, since in this area measured concentrations are supposedly lower. Furthermore, he recommended us to assure the quantification of the signal and preferentially the concentrations as well.

Onno Epema from Rijkswaterstaat (part of the Dutch Ministry of Infrastructure and Water Management) pointed out the importance of pharmaceutical waste products in water, for which our system could improve the detection.

Surface water contaminants contain a wide variety of compounds, all with a different ecological impact. The meeting with Martin van den Berg, Deputy Director of the Institute Risk Assessment Sciences (IRAS), guided us to focus on one of the forthcoming most hazardous emerging contaminants: Selective Serotonin Reuptake Inhibitors (SSRI). These hormone mimicking compounds are used as antidepressant, and include pharmaceuticals such as Prozac. Even in low concentrations SSRIs are pharmacological active in vertebrates and have an ecological impact.

The interactions with both stakeholders and experts provided us with a number of applications for our biosensor, such as a pre-check for the chemical analysis and local sampling. To enable the use of DeTaXion to these ends, multiple requirements had to be integrated into the applied design. Key values for this purpose were that the device has to be quick, safe, accurate and quantitative. More information on our applied design and the integration of insights of experts and stakeholders can be found on the Applied Design page.

Ethical issues among GMO’s and Detaxion

The use of GMO has increasingly come under scrutiny in a range of different fields, with public resistance mainly focussed on its appearance in our diet. Herein, the application of the GMO matters: use of GM animals for the production of pharmaceuticals is viewed far less negatively than use for food (PEGASUS. (2012). Final Report - PEGASUS (Public perception of genetically modified animals - science, utility and society). European Commission CORDIS, United Kingdom). Being a subject of strong debate, it is important to ascertain these ethical concerns for our project and product design, as our biosensor naturally is a GMO. As our sensor is best utilized directly in the field, it is implied that our genetically modified bacteria leave the laboratory to enter the outside world - safely contained within our device - leading to the primary concern of introduction into the environment. This introduction of a modified bacterium is potentially harmful, as it could disrupt ecosystems, introduce antibiotic resistances in other species, and have harmful health effects on humans. These concerns were echoed by Martin van den Berg (deputy director of the Institute of Risk Assessment Sciences), who stressed the importance of containment of our E.coli within our device to prevent introduction. Our device therefore contains a number of safety measures to prevent this, such as adequate sealing and storage, and stringent cleaning and washing steps. For more information on how we integrated these safety issues into our applied design, visit our applied design page. Bacteria themselves also contain a number of safety measures: it is firstly comprised of a weak lab strain, with poor survival chances as a result. The bacteria also contain a killswitch-system preventing survival outside of the device. With this approach, we ensure safe use of our device outside of the laboratory.

Ultimately, utilitarian ethics plays a large role in the public perception of GMO. If the public is sufficiently aware of the urgency and importance of achieving a specific goal, it will be more likely to be open to the use of GMO (Hersh, M. A. (2002). Ethical analysis of automation: a comparison of different ethical theories through case studies. IFAC Proceedings Volumes, 35(1), 351-356.). In the case of our biosensor we theorize this means that the public is likely to be more open to the use of GMO, as the potentially harmful effects of pharmaceutical pollution are not currently being addressed sufficiently. Multiple experts confirmed this sentiment: as long as the genetically modified bacteria in our device are safely contained, a majority of people will not oppose their use due to their important utility in solving the detection problem of pharmaceutical pollution (Martin van den Berg, Institute of Risk Assessment Sciences). In a similar vein, our biosensor makes current detection methods (i.e. those utilizing Daphniae and fish) redundant, leading to a reduction in the use of laboratory animals.