One of the pillars supporting a good and responsible iGEM project is human practices. We prioritize human practices to ensure the real-world effectiveness of our product and access the impact on Danish society, globally, and the scientific community.
Therefore, we aim to address the fundamental iGEM human practice questions:
How does our project influence the world around us? And how does the world around us influence our project?
Our goal is to prevent PFAS from causing harm to humans and the environment by removing the hazardous chemicals from our drinking- and wastewater. This is achieved through our vitroZymes, bioengineered enzymes that degrade PFOA molecules. To grasp the extent of the PFAS issue, we engaged with our community.
We explored citizens’ concerns, sought the perspectives of Danish politicians on biotechnology, and discussed our project with experts in PFAS pollution, as well as larger companies involved in water-related industries to gather their insight for a more sustainable solution for PFAS disposal.
Our goal was to incorporate a broad spectrum of viewpoints from Danish citizens to larger companies to consider all possible perspectives that our project might entail.
Now you can choose to read about the people we have worked with, or if you want to delve into our considerations about the concerns of PFAS and GMO in society. Clicking this button will hide the other section.
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To understand the potentially far-reaching impact of our work on the community and the environment, we visited various waterworks and consulted experts in water quality, GMOs, and ethics.
Our journey began with the understanding that clear and accessible communication is essential when dealing with a complex topic like PFAS. We discovered that there is no one-size-fits-all substitute for PFAS, but alternatives to PFAS are slowly emerging in consumer products and the industry. Through our conversations with experts in the field, the environmental and health costs associated with PFAS contamination were highlighted. They emphasized the urgency of finding an effective and sustainable solution, and we hoped that our enzymes could provide that solution.
To understand how to implement our solution industrially, we visited waterworks and learned about how their functioning water filtration systems work. The simplicity of their water filtration methods inspired our own approach. We discovered that effective solutions can often be straightforward and efficient.
These insights have deepened our understanding of the PFAS problem and have guided our efforts to develop a solution.
Natascha Dimple - Philosophy and Ethics
Why did we establish contact?
We wanted to gain a deeper understanding of the ethical considerations around the area of synthetic biology, laboratory work, and GMOs. We wanted to gain some tools for ethical analysis and awareness. Therefore, we contacted 2021 SDU iGEMer Natasha Dimple who has a bachelor's degree in philosophy and is doing a master’s in engineering. During their time in iGEM, Nastasha Dimple’s team made an extensive ethical analysis based on their project description.
What we learned
We should consider the desired influence we want our iGEM project to have.
We should keep evaluating if our product benefits the world and what potential harm the fluoride by-products could have.
The importance of using the right terminology and analytic strategies when discussing our ethical considerations.
Reflection
Natasha Dimple was a great choice for a guide to the understanding of ethical considerations. Since she has experience analyzing the ethical aspects of an iGEM project, she had some very relevant pointers for us. We found the analytical strategies she suggested helpful when building the structure of our ethical analysis.
Philippe Grandjean – Professor in Environmental Medicine
Why did we establish contact?
Professor Philippe Grandjean is a prominent figure in the field of PFAS, renowned for his research on long-term exposure to pollutants at Harvard University and the University of Southern Denmark. We were eager to learn about his findings and the observations he has made throughout his lifelong dedication to this field. This knowledge would help us gain insight into the side effects of PFAS, and how this problem started.
What we learned
The chemical structure of PFAS molecules resembles fatty acids.
Phillipe mentioned that exposure to PFAS-molecules can reduce our antibody production by up to 50%.
Large PFAS manufacturers, such as Dupont, hid their discoveries about the harmful effects of PFAS, resulting in a large-scale scattering of PFAS across the country. These companies are now facing huge lawsuits9,10.
He went on to explain that production of both PFAS-precursors and branched PFAS continues in Chinese products.
Misinterpretation of data from one of his studies by the Danish government has resulted in the PFAS legal limit in drinking water being set too high. Based on his calculations, the limit should be set even lower.
The Uppsala Commission released data that showed that the societal cost of removing a PFAS product is 1000-fold higher than its purchase price.
Reflection
After our conversation with Philippe, we contemplated the significant environmental costs of PFAS. We reviewed the articles he provided, which proved to be quite informative.
We also contemplated the origin of this problem and began to brainstorm ways to raise awareness of PFAS. Many of the issues could be mitigated if we, as humans, ceased production of these persistent chemicals. Both the societal costs of PFAS and the associated side effects reinforced our perspective on the PFAS problem and the urgent need for ways to solve it.
Bioomix
Why did we establish contact?
We contacted this company because they utilize microorganisms for agricultural improvement. Our purpose was to know their approach to GMO regulations. In that regard, we wanted their input on our product, and to explore potential areas in which we could improve. The CEO and Co-founder of Bioomix, Morten Andersen, kindly provided us with some advice.
What we learned
Our enzymes might struggle to bind to PFOA molecules due to their relatively low abundance.
The slightest change in releasing GMOs, especially transgenic bacteria, into the environment would not be approved by GMO laws.
Reflection
Our primary concerns regarding Morten’s comments centered around navigating GMO regulations and finding ways to degrade PFOA using biotechnology. We considered isolating enzymes and incorporating them into a filter to bypass the challenges that live-cell systems pose.
As for the problem of low PFOA abundance, we decided to optimize our dehalogenases at high PFOA concentrations and reach out to companies that work with PFAS to try our enzymes on some water samples. Unfortunately, our vitroZymes were not ready to be tested before the deadline.
Vandcenter Syd - Drinking Water Department
Why did we establish contact?
To get the angle of our local PFAS problem, we contacted VandCenter Syd. A local waterwork in Odense. It is one of the oldest in Denmark and was dug by hand in 1853. We contacted them because we wanted to understand the work that is already implemented in providing clean and safe drinking water to our city.
What we learned
There is yet no PFAS in Odense groundwater. However, VandCenter Syd considers the arrival of PFAS in the groundwater as inevitable. Ulla Brinkmann Trettenes, a specialist in drinking water quality, said, “It’s only a question of when and not if PFAS will show up in our drinking water”.
The waterworks filtration system is based on an almost 150-year-old method, where unwanted heavy metals in the water are filtered out by different types of sand and rocks. The natural bacteria in the sand and rocks remove chemicals such as ammonium from the water.
We should reach out to the VandCenter Syd wastewater department to hear more about PFAS in Odense.
Waterworks are interested in looking into any viable solution for the PFAS problem.
Reflection
The Danish waterwork companies pride themselves in the simplicity of the Danish method of cleaning water wherein we can pump the ground water up and run it through sand. They understand the extent of the PFAS problem and seem willing to try new solutions. The insights into how the filtration system works helped us understand some of the obstacles we had to face for our project to be ready to implement. We were very interested in their reasoning behind the different sand types because we saw an opportunity to insert our enzymes.
Tune Waterwork
Why did we establish contact?
Susan Münster from The Association of Waterworks in Denmark, whom we met at the People’s Meeting on Bornholm, had informed us about the PFAS challenge at Tune Waterwork We connected with Kurt Ardal Larsen, the operational manager at Tune Waterworks, who has been working on a solution for the PFAS problem for the last two years.
What we learned
Tune Waterwork struggles with high concentrations of mainly PFOA and Perfluorohexane sulfonate (PFHxS) in nearly all their boreholes.
Drilling just one new borehole costs a staggering 1.2 million DKK (approx. 174,000 USD).
Kurt told us about Silhorko, a company designing new water treatment solutions. Silhorko was the first to develop a PFAS filter, which they tested at a waterwork on Fanø.
Kurt also told us about Split Water Nordic filters, a company also working with water treatment solutions. They make everything from membrane filtering to Pulverized Active Coal (PAC) carbon filters.
Reflection
The shortcoming of these solutions was that they only filtered the water for PFAS, leaving the problem of degradation. The disposal of PFAS-accumulated water remains expensive.
We got blown away by the extortionate cost of drilling a new borehole. We saw the need for an easier and cheaper solution to clean the already existing boreholes, which we decided to work towards. From this conversation, we wanted to make a cheap PFAS degradation process, which could be used even by small waterworks or companies. Read more about this later.
We were also confirmed in our decision of choosing to focus on PFOA over other PFAS, as it was the main PFAS found at Tune Waterwork.
Frederikshavn Forsyning
Why did we establish contact?
We arranged a meeting with Rasmus Pedersen and Kenneth Fuglsang from Frederikshavn Forsyning. They manage a waterwork in the Danish city, Skagen, in Northern Jutland. At the boreholes in Skagen high concentrations of PFAS have been found. We wanted to learn about how they tried to resolve the issue and start a conversation about potentially implementing our synthetic biology solution.
What we learned
However, due to PFAS contamination, Frederikshavn had to shut down their borehole in Skagen.
The region is gradually converting to being supplied by other wells in Northern Jutland.
The resin filters were too expensive to implement at Frederikshavn vandværk.
The remaining waterworks are diluting the PFAS concentration from contaminated water by mixing the water from different non-contaminated boreholes.
The waterworks were captivated by the notion of biological degradation. They regarded it as a fantastic concept due to its sustainability and potential cost-effectiveness.
Towards the end of the talk, Kenneth said the following:
“It might be, that what we're seeing right now is just the tip of the iceberg – as our drinking water is currently 50 years old, but what will it look like when we reach the water which is younger?”
Reflection
After this meeting, we reflected on the fact that PFAS-contaminated water is diluted in our drinking water. We believe that this method is unsustainable in the long run. Given the scope of the PFAS problem, we believe that one day we will not have any clean water to dilute it with. This poses a problem that we hope our solution can help with.
Rikke Markfoged from the Danish Technological Institute
Why did we establish contact?
PFAS has become a source of anxiety and confusion for the average Danish citizen due to the way the topic has been extensively covered in the media. We consulted Rikke Markfoged, Senior Project Manager at the Danish Technological Institute, who has been featured in Danish media discussing PFAS. We wanted to get a better understanding of how PFAS is discussed in her work with waterworks versus how it is communicated to the public.
What we learned
When discussing a topic with many unknowns, such as PFAS, it is important to stick to the facts.
The public takes notice of shocking stories and creates a demand for more information.
Awareness about PFAS informs consumer choices.
Media coverage makes politicians take notice of the issue.
Accessible and accurate information is key to effective science communication.
Reflection
Rikke emphasized the importance of making information accessible during science communications to not scare the public too much, but still get the point through. We considered Rikke’s advice every time we mentioned PFAS to the public going forward.
Vandcenter Syd wastewater
Why did we establish contact?
Unlike in drinking water, PFAS is found in Odense’s wastewater. We wanted to discuss the issue of PFAS with employees at the wastewater department at Vandcenter Syd. We spoke with ecological biologist Rikke Hansen, wastewater manager Ivan Vølund, and laboratory team leader Lise Havsteen.
What we learned
The current methods for cleaning wastewater are very similar to the methods used for drinking water. Vandcenter Syd uses sand filters, aerate the water, and naturally occurring bacteria to remove waste products and metals from water.
VandCenter Syd measures 22 types of PFAS in their wastewater, even though they expect many more types of PFAS in their water.
Removing PFAS from wastewater might be more effective than drinking water because the cleaning process is slower and more extensive.
Reflection
From our visit to Vandcenter Syd, we gained insight into how the water filtration systems worked and how they had to treat drinking water and wastewater differently. We contemplated focusing on removing PFAS from wastewater, instead of drinking water, but we determined that our solution could potentially be implemented in both types of processes.
HOFOR
Why did we establish contact?
Hovedstadsområdets Forsyningsselskab (HOFOR) is the utility company that serves the capital region of Denmark. From our visit to Vandcenter Syd, we knew that HOFOR had already experienced PFAS-contaminated drinking water and we were therefore eager to understand how they handled the PFAS pollution. To achieve this, we reached out to Liselotte Clausen, the Chief Consultant at HOFOR.
What we learned
HOFOR has previous experience with a former DTU iGEM team, where they employed live bacteria to produce enzymes for pesticide removal in wastewater. However, the attempt stalled as natural bacteria outperformed the gene-modified bacteria over time.
Liselotte is open to synthetic biology approaches. She saw potential in our idea of using GMOs to clean PFAS-contaminated water.
Using a resin filter to remove PFAS could introduce resin contamination in the drinking water. Curious about this, we reached out to Silhorko, the creators of the resin filter, to gather their perspective on this take. Read more later.
Liselotte raised a concern that if a lot of cleaning and filtration systems were implemented, there might be less protection of our natural resource, groundwater.
Reflection
After learning that GMOs are outperformed in sand filters, we decided to find another way to integrate our enzymes into the water treatment process. We started looking into other parts of the process where we could implement our solution.
She gave us a reason to reflect on how to promote PFAS degradation solutions while advocating for the protection of our groundwater. Implementing a lot of filtration systems could potentially lead to even more contamination of water due to the lack of care. We want to make sure that in the branding of our solution, we present GMOs as an alternative to unsustainable incineration. PFAS production and consumption would remain unethical and unsustainable for humans and the environment.
Emil Damgaard-Møller from the Danish Technological Institute
Why did we establish contact?
Emil has been a prominent person in Danish media regarding the PFAS problem. We contacted him to discuss his insights into alternatives to PFAS. He was happy to work with us, and we met up with him at the 2023 People’s Climate Meeting in Middelfart. Read more about the event here.
What we learned
There is not one single substitute for PFAS, but new substitutes are beginning to develop.
Example: Some raincoat companies have begun using Polyurethane as a waterproof substitute.
Many areas in infrastructure and production have alternatives available so companies must start making the conscious decision.
Degradation of PFOA might release a harmful fluorine gas, but according to Emil’s past experiences, the gas would be easy to remove before it would become damaging. But it is something to consider.
Reflection
Our goal is to be able to degrade PFAS molecules, but without releasing another harmful pollutant. This conversation prompted us to investigate the byproducts of PFOA degradation. We specifically looked into how to dispose of the fluorine gas.
SDU RIO
Why did we establish contact?
The University of Southern Denmark (SDU) Research and Innovation Organization (RIO) is an organization that aids students with their business start-up dreams. We wanted to establish contact with SDU RIO to learn about Development and business ideas for a project like ours. We wanted to get a better understanding of how to develop pitches and presentations, apply for grants, and develop a prototype.
What we learned
The importance of a hook, visual aids, and speaking to the audience during a pitch.
Visual aids, such as a prototype, can help the audience grasp the idea.
Going to events and trying off our pitch is great practice.
Reflection
We learned through our discussions on how to make a pitch, that altering the level of detail when explaining synthetic biology would always be necessary to accommodate our audience. This is essential to keep the audience focused. We started developing ideas for visual aids to help the audience understand the PFAS problem and our product.
Silhorko - Fanø waterwork
Why did we establish contact?
Silhorko is a company that specializes in water filtration systems and implementation in water treatment facilities. We came across Silhorko in the news at the beginning of our project, where we learned they had developed a new PFAS resin bead filter on Fanø. We set up a meeting with Arne Koch from Silhorko and Jan Pedersen from Fanø Vand at the Fanø waterwork during a filter maintenance check. We were intrigued to gain insight into how the resin filter worked, the extent of the PFAS problem at Fanø, and their thoughts on the possibility of using GMOs to degrade PFAS from their filters.
What we learned
Silhorko’s filter was developed using resin chloride ion exchange technology.
Silhorko’s resin beads have a chloride group that is secreted into the water. PFAS replaces the chloride group and binds to the resin, thereby capturing the PFAS from the water.
The resin filters could operate with a flow rate of 250 liters of water per hour and they assured us that no signs of PFAS were detectable afterward.
The filter's lifespan is still under review. Current estimates of the life span are 2-3 years.
Arne is optimistic about implementing enzymes like our vitroZymes to extend the resin filter's longevity.
Fanø Vand has implemented two new filter tanks for PFAS filtration, each containing 4500 liters of resin. The resin cost approximately 900,000 DDK, (approx. 130,000 USD).
For safety purposes, before they started using the filters a contamination test was conducted to ensure no toxic chemicals were released. During this test, they detected a small amount of styrene, which Arne Koch assured is a molecule typically found in resin filters. He explained that it can be resolved by rinsing the filter a few times before use.
They provided us with some resin chloride beads, which we could use for a prototype, and the contact information of Søren Duch-Hennings, a chemist and former employee at Silhorko, who worked on the resin filter.
Reflection
We began concocting ideas for the design of a vitroFilter. We wanted a solution that built upon the resin filter.
We were inspired by the resin filter we had seen at Fanø. We decided to implement the beads they gave us in the prototype of our vitroFilter to demonstrate how our enzymes could be incorporated into real-world filters. Unfortunately, during the design process, we realized that our enzymes worked better as a filtration step rather than a physical filter. Read more later in the integration section.
Søren Duch-Hennings
Why did we establish contact?
Our conversation with Silhorko on Fanø led to Søren Duch-Hennings, a former Silhorko employee who worked on the development of the resin filter. During our discussions, we gained a deeper understanding of how the resin filter was constructed, including its chemical aspects.
What we learned
The resin filter is a Disposable solution, meaning it is not designed to be reused.
Silhorko uses chloride beads, to catch PFAS in a solid phase rather than a liquid phase.
PFAS is soluble in ethanol. This property can potentially be a way to release PFAS from the chloride beads.
There are alternative methods of accumulating PFAS, such as nano-filtration through membranes or the utilization of surface activation foam fractionation (SAFF).
We must follow the Drinking Water Executive Order when developing new water filtration technologies.
Reflection
The primary focus of this conversation centered around the implementation of our enzymes. We investigated the alternative methods for PFAS filtration to identify where our PFAS degradation solution would make the most sense to implement.
On a global scale, ensuring access to clean drinking water for all is a fundamental aspiration. However, access to clean drinking water is considered a human right, this core belief is now challenged by the presence of PFAS. Our team firmly upholds the principle that everyone should have access to safe drinking water. We greatly value our ecosystem, environment, and the health of all beings. All of which are greatly dependent on clean water.
The relevance of the problem was quickly confirmed at the People’s Climate Meeting in Middelfart, which we went to at the beginning of our project. The People’s Meeting on Bornholm gathers scientists, politicians, and regular citizens to talk about the relevant problems at the time. There was quite a lot of focus on PFAS, and we went to different discussions on the PFAS problem, where we gathered information and contacts. At an event arranged by the supermarket Coop, we learned that of their customers who were familiar with PFAS and participated in the survey, 67% were worried about it. That only confirmed our cause. At the event, we spoke with stakeholders from waterworks who were very interested in our solution. We spoke with politicians like the Environmental Minister of Denmark, Magnus Heunicke, about our solution, and when we asked him if he saw the potential in PFAS degrading enzymes, he said “Definitely”. Overall, the substantial attention on the PFAS problem suggests that our solution is well-timed and aligns with current priorities.
The idea of solving this problem started with input from team members’ families, who were worried about the chemicals in drinking water that are harming humans. PFAS’ harmful effects have been studied for many years and scientific studies are only increasing in amounts1. However, we found that knowledge about PFAS was not common outside the scientific environment. Before starting our iGEM project, our own team members were unsure of what the toxic substances were. At our education session at Nordfyns Højskole, only one person in a class of 30 people had heard about the PFAS problem before. This raises the question of whether it is relevant for citizens to learn about the harmful effects of PFAS. Read our thoughts in the Ethics section.
The Fear of Genetically Modified Organisms
Using GMOs for our solution comes with considerations of how the discourse in society affects our project. GMOs have been associated with fear and have historically been presented as unnatural in the media. We aim to alleviate these concerns with effective communication and information sharing.
GMOs are subject to strict regulation under the European Union's GMO legislation2, ensuring safety, transparency, and traceability.
The EU’s GMO legislation is a legal framework that ensures that the development of GMOs and biotechnology in general are done in safe conditions. They have high standards for GMO-containing products before they are introduced to the market. They also have risk assessments and authorizations of GMOs that are efficient, transparent, and easily understood. The EU makes sure that when GMOs go onto the market, they are traceable and clearly labeled, so consumers can make an informed choice of what they buy2.
Cognitive Bias against GMOs
In our conversation with former iGEMer Natascha, a graduate student in engineering with a bachelor's in philosophy, we discussed that many citizens and people who do not work in biology-related sciences might have a misguided idea of what GMOs entail. People might have a cognitive bias on the use of GMOs. Cognitive bias is when a person subconsciously overlooks one kind of information and in favor of another. Therefore, a part of the population might be influenced by media that produces fear of GMOs and ignore the science-based evidence that promotes the use of GMOs.
We also discussed pro-science deniers, people who are science supporters but deceit themselves, when science tells them something against their personal beliefs or values. In this instance, people might ignore the scientific research and statistics that show the dangers of PFAS, because they fear having to change their habits or the reality of how dangerous an invisible chemical can be.
These are aspects we have to keep in mind when communicating about our product.
Ethics
At vitroFAS, we conduct biotechnology research staying within the bounds of GMO laws and following laboratory safety regulations. Furthermore, we recognize that ethical and responsible scientific research does not only involve experts. To create impact, we must engage and involve citizens to make our ideas applicable to real-world challenges. We believe we must address people’s concerns about toxic chemicals and synthetic biology solutions.
Read more about our argument in the dropdown below.
In the discourse surrounding PFAS pollution and its associated side effects, ethical considerations play a pivotal role in determining whether the public should be informed about these issues. At vitroFAS, we want to be transparent about the various ethical perspectives on the topic.
We believe it is ethically imperative that people should have access to knowledge about PFAS pollution and its potential repercussions.
Therefore, we evaluate this perspective based on the term consequentialism. It is the judgement of actions based on the consequences of the action rather than the action itself.
The Case Against Sharing Information on PFAS
One argument against widespread knowledge of PFAS pollution lies in direct consequentialism. The Stanford Encyclopedia of Philosophy defines the ethical standpoint of direct consequentialism as whether an act is morally right depends only on the consequences of that act itself3. From this perspective, disseminating information about PFAS may only add fear and concern in the modern citizen who is already burdened with concerns about other issues such as the economy and the climate crisis.
Another argument against sharing information stems from the aggregative consequentialist perspective, that evaluates which consequences are best in some function of the values of parts of those consequences3. The idea that a vast number of people would need to become concerned about PFAS pollution before tangible effects, such as resource allocation for research or legislation, can be realized, raises concerns about potentially fearmongering the public. To avoid fearmongering, vitroFAS must stick to making accurate information accessible to the public. We have done so by reading research papers before putting information up on the Wiki and communicating the information at events.
The Case For Sharing Information on PFAS
What direct consequentialism and aggregative consequentialism fail to consider is the long-term effects and intentions of sharing information on PFAS. These shortcomings are offered in universal consequentialism, a perspective that evaluates moral rightness by the consequences for all people[3]. By ensuring that a smaller population is well-informed, we can initiate research and legislation that would benefit not only the present generation but also those to come. In the long run, the positive effects of informed decision-making significantly outweigh the immediate concerns. The longer the topic of PFAS pollution remains under wraps, the more PFAS pollution, and the bigger the consequences.
Furthermore, from an evaluative consequentialist standpoint, sharing our findings and the lessons we learned on our iGEM journey contributes to the collective pool of knowledge on the topic of PFAS and synthetic biology. The resources, time, and effort invested in the vitroFAS project will support the scientific community. This, even if we don't produce a tangible product, has the potential to inspire further research and innovation in the field of synthetic biology, as well as the development of groundbreaking solutions to mitigate the effects of PFAS pollution.
Lastly, we must evaluate the deontological perspective, which criticizes consequentialism for being able to justify any kind of act, for it does not matter how harmful it is to some so long as it is more beneficial to others4. Following deontological ethics, which emphasizes listening to our morals, we must share our knowledge. Having worked extensively with PFAS, we feel a moral obligation to address the consequences of its pollution, despite not being directly responsible for it. This sense of duty extends to rectifying the wrongs perpetrated by large companies and advocating for change.
In conclusion, a careful examination of the ethical dimensions surrounding sharing knowledge about PFAS pollution and its side effects, strongly suggests that it is not only beneficial, but also ethically imperative to disseminate this information. From various consequentialist perspectives, it serves the greater good, scientific innovation, and the duty to act morally to share what we have learned with the public. By doing so, we empower citizens to make informed choices and pave the way for a synthetic biology solution for PFAS pollution.
vitroFAS aims to address the concerns of PFAS pollution not only in the water treatment industry, but also among the public. We attended public events and schools to introduce citizens to the nuances of the PFAS problem and our project. We want to promote public awareness about innovative science approaches, such as our vitroZymes. Our biotechnology solution addresses concerns about the energy-consuming and ineffective practice of incineration for PFAS degradation by suggesting an alternative synthetic biology solution.
Demystifying GMOs and PFAS
Involving the opinions of the everyday citizen inspires us to work on science communication. By demystifying the technology and addressing misconceptions, we can bridge the gap between scientists and the public, encouraging support and trust for responsible research practices.
Engaging with the public, we found varying opinions on GMOs.
We visited the school Nordfyns Højskole, where we asked the students about their opinions of GMOs. None of the participants had any strong opinions or fears of GMOs. We also visited the People’s Climate Meeting in Middelfart, where people were split on this issue. Some did not like using GMOs to solve the PFAS problem, while others encouraged it. This might indicate a shift in public opinion about GMOs.
We also went to the People’s Meeting on Bornholm, where we had the opportunity to ask politicians and water experts their opinions on using bacteria to remove pollution and other synthetic biology endeavors. Once some of the citizens and politicians we talked to, heard examples of how far biotechnology has come and its current uses, they became excited about the field’s prospects.
To learn more about the PFAS problem, we talked with several waterworks, some of our most important stakeholders. From these conversations, we realized that PFAS is a major problem in Denmark. In a data sheet provided to vitroFAS by Danish Waterworks, 67 out of all 224 waterwork facilities in Denmark found levels of PFOA, PFOS, PFNA, and PFHxS contamination above the limit value in their raw water (groundwater and other water bodies)6. These elevated levels of PFAS molecules led to the shutdown of several facilities.
Past PFAS degradation methods include incineration, ultrasound degradation, microwave thermal treatment, supercritical treatments, thermal desorption, and vapor energy generators (4). Most of these methods are expensive, use large amounts of energy, and have inadequate results. For instance, incineration can release other types of PFAS into the environment as secondary pollution because the process degrades PFAS incompletely6,7,8. These approaches lack the innovation of synthetic biology.
The waterwork representatives are eager to find new solutions because current methods to remove PFAS are largely inefficient and impossible to implement on a large scale. We learned that should a viable alternative arise; the industry would be incentivised to cover the cost. The waterworks would no longer risk being shut down due to PFAS-contamination of their water if a solution like our vitroZymes was implemented.
We discovered that the waterworks were open to introduce biotechnology at their facilities. Larger waterworks, such as HOFOR and VandCenter Syd had previously collaborated with research groups and introduced GMOs among their natural bacteria. Read more in the integration section.
Integrating Stakeholders In Our Device Development
A solution for individuals, companies, or the state?
Through the development of our project, we reflected on who the consumer for our project would be. At one point, we considered a business-to-consumer model where our filtration system would be implementable in household faucets. Our team moved away from this idea because we agreed that it should fall on the government to ensure safe drinking water for its citizens. It is a human right to have access to clean water that encompasses water without PFAS. Therefore, we decided our solution should be as close to the source of PFAS contamination as possible, and that is how the idea of implementing our enzymatic step in waterworks, a business-to-business model, came to be. This would serve the population countrywide rather than forcing individual households to decide whether to invest in a PFAS filter and degradation system in their homes. A large-scale solution like ours also discourages more consumerism than single household applications, making it more sustainable.
Implementing our system into current water filtration platforms.
We have explored various potential methods for integrating our technology into the real world when the GMOs are successfully optimized for PFAS degradation.
Our first idea was to use a “submarine-like” device with live cells in an enclosed system that we wanted to submerge in water bodies like lakes and rivers contaminated with PFAS. Our logic was that if we could clean water bodies from PFAS, then it would not spread to the groundwater that we drink. While acknowledging the benefits of using GMOs in bodies of water, we recognize that a closed system offers a safer and more straightforward implementation approach. Such a system reduces the risk of unintended spread and interactions with local wildlife, thus lowering ecological risks.
Our goal was originally an E. Coli bacterium that could both absorb, degrade, and repurpose PFAS and its fluor contents. We, however, quickly realized this was very ambitious for the timeframe we had and decided to focus on the aspect of the project we found most important. To narrow it down we went with degradation of one of the most widespread and studied types of PFAS: PFOA.
Why PFOA?
Because there are thousands of PFAS types, we narrowed our focus to PFOA, one of the most widespread and researched molecules of PFAS11. The USAFA iGEM teams research, which was where we discovered the dehalogenases, used PFOA as their model substrate
Our talk with Tune Waterwork confirmed our choice, as their biggest problem was with PFOA. In our modeling work, we found that other types of PFAS possibly have a higher affinity for our enzymes than PFOA, so the vitroZymes might be able to degrade a larger range of PFAS types. Read more about that on the Model page.
We consulted the company Bioomix, who has experience with implementing GMOs into farming for agricultural improvement, for their insights on navigating GMO laws and regulations. They informed us that the current laws for GMOs use in ecosystems would prevent us from implementing this solution.
Our next implementation draft was to integrate our synthetically modified bacterial strain into the existing bacterial population in water treatment facilities. Through our dialogue with HOFOR and Vandcenter Syd, we learned that introducing new bacterial strains into existing populations often outcompeted the new strains, so we went back to the drawing board.
We had to readjust our implementation strategy, and we decided to build a filter that would only contain our purified dehalogenases.
The vitroFilter
Before designing this filter, which we named the vitroFilter, we needed to understand how the filters at treatment plants work. We met with five individual water treatment plants across Denmark to learn how each of them handles the PFAS in their water and learn more about their filtration method. We learned that all water treatment plants in Denmark use a sand-based ion exchange filtration system developed over 150 years ago. This ion exchange system removes metals from the water using bacteria in the sand.
Our dehalogenases would not be effective if implemented directly in the sand filters at waterworks. The utility company HOFOR, which supplies the capital region of Denmark, collects approximately 50 million cubic meters of groundwater for drinking water annually12, which means that HOFOR alone filters over 2000 tons of water per hour at their facilities. Liselotte from HOFOR explained that some boreholes at their Solhøj facility have PFAS concentrations above the drinking water limit13.
We want to get the most out of our enzymes. We determined that our vitroFilter would be more effective in water bodies with higher PFAS concentrations. So, we needed another solution.
Cleaning the resin bead filter
Fanø Vand tested to see if the resin filter was as effective as the carbon filters normally used for extracting PFAS. They found the resin filter to be more effective and less costly than the carbon filters other waterworks use. Despite removing PFAS more effectively from the water, it cannot degrade the chemicals, which are instead sent for incineration. Therefore, our proposition of developing an enzymatic degradation process was met with great enthusiasm.
Because the waterworks already have a means of collecting PFAS, our idea is to clean the saturated PFAS resin filters, so they can be reused. We would wash the resin filter with a mix of ethanol and water to release the PFAS from the resin beads. We were suggested an ethanol mix due to the mostly hydrophobic property of PFAS, but we were recommended to add in water to lower costs. The water would now be contaminated with a high concentration of PFAS to be degraded by the vitroFilter.
Scrapping the vitroFilter
In the end, we decided that our enzymes alone were the product of our iGEM project. The resin bead filter and other PFAS extraction methods could concentrate PFAS, so our vitroZymes can work in high concentration substrate environments.
Our final idea is to collect highly concentrated PFAS water in a container. This will allow PFAS collected from various filtration methods to be workable in our design. For example, we can elude PFAS from the resin beads by rinsing them with an ethanol/water mixture and pumping it into our container. Next, we will insert the vitroZymes into the water and allow it to degrade the PFAS. Then, the water will be boiled to denature the enzyme and we remain with water with fewer PFAS molecules. Read more on our Implementation page.
These developments of our product would not have been developed without the aid of experts in their respective fields. The help we gained was essential and matured our project from an initial idea into a product that could be implemented in the Danish water filtration system.
How will our project improve the world?
The vitroZymes are a sustainable synthetic biology alternative to degrading PFAS with enzymes. To make the solution implementable, our enzymatic process has been developed to be easily incorporated into current water filtration systems.
Final thoughts
Our meetings with the waterworks helped us identify the water treatment industry’s needs and interest in our synthetic biology solution. We learned more about how to implement our product in the real world.
The inspiration, guidance, advice, and improvements we have gained through our outreach and human practices work, have been invaluable to our project. We’ve gained insights into the professional world and become better at critical thinking, communication, and organizational skills.
While the mission of our team to remove PFAS from water bodies has remained the same, our goals and approach have been adjusted along the way. These changes were based on new knowledge and experiences that altered our final product to a more sustainable, responsible, and pragmatic solution.
Our final product is the result of the great advice we have received along our human practices journey. We are proud of our product and all that we have learned through our iGEM endeavor, both as individuals and as a group.
Value Proposition Canvas
Fig 1: This is the value proposition canvas of our vitroZymes as a solution for waterworks.
Above figure depicts a Value Proposition Canvas for our customer, waterworks, to evaluate our solution to their PFAS problem. We consulted with Dennis Nielen de Raadt, studying BEng in Global Management and Manufacturing at the University of Southern Denmark.
The Value Proposition Canvas is a tool, a visual framework, commonly used by organizations and businesses to design and improve their products or services. The canvas helps understanding and aligning the products, with the needs and preferences of customers.
In the following, we look beyond the final development of our product and the impact of the final product.
Customer jobs ↔ Products & Services
The jobs the customers need are the filtering and disposal of PFAS. Our PFAS-degrading vitroZymes can be combined with existing filter solutions, degrading concentrated PFAS water from the filters.
Pains ↔ Pain Relievers
The waterworks risk being unable to follow new regulations and getting a bad reputation if they keep disposing of PFAS in an unsustainable way. New and more stringent regulations to prevent PFAS from harming people and the environment are continuously being implemented. Regulations force the waterworks to implement a solution quickly, which has the risk of being very expensive14.
Waterworks know PFAS will increasingly be a problem in their boreholes in the future. They also know that incineration has been shown to release PFAS back into the environment, extending the lifecycle of PFAS, which has to be processed by the waterwork again. If waterworks do not look for a more sustainable solution, they risk dissatisfying customers and therefore stricter regulation.
The demand for a PFAS degradation method is high.
In combination with a filter, our vitroZymes will prevent the outlet of PFAS into the environment and proactively address future complications. The waterworks will also keep the customers satisfied by taking action against PFAS.
Gains ↔ Gain Creators
The waterworks wish to label themselves as sustainable, innovative, and responsible to keep their customers happy. Meanwhile, they want to keep their operation as simple as possible. Investing in our vitroZymes gives the waterworks the ability to brand themselves as proactive and innovative against the PFAS problem.
As of right now, we cannot fully account for the simplicity of our vitroFAS solution. We do not yet know the details of how to dispose of the fluoride by-products. A future perspective of this project is to look into how to provide an on-site solution that can easily be implemented at the waterworks.
Avoid greenwashing
Although the vitroZymes are not yet ready for commercial production, we are conscious that biotechnology solutions can be used to greenwash organizations and companies.
vitroFAS is committed to being transparent about our product.
Since the vitroZymes currently have only been tested in their ability to degrade the molecule PFOA, we cannot allow water work companies to promote our product as a complete solution to removing PFAS from the environment as of right now. We need to underscore that vitroZymes only have the potential to degrade other molecules, but further investigation is required.
Most importantly, we want to make sure that the vitroZymes solution is not an excuse for PFAS manufacturers to continue unnecessary PFAS production.
Conclusion
In recent years, the PFAS issue has been a hot topic among politicians, scientists, and water utilities – all of whom are looking for a new, sustainable solution for disposing of PFAS.
PFAS have been researched for many years, yet they remain relatively unknown to the public. We found it most ethically correct to tell the citizens about the harmful effects of PFAS, so they have the knowledge and opportunity to make informed choices on PFAS. We acted on this by educating young minds, speaking with stakeholders, and communicating our knowledge at public events.
Although it is still in its beginning phases, the vitroFAS solution has been met with great interest because it has come to our target customers’ attention at the right time and has the potential to solve many of their needs. Our solution has developed into a product that can be implemented in the real world.
Education
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European Commission, E. GMO legislation. https://food.ec.europa.eu/plants/genetically-modified-organisms/gmo-legislation_en#further-information
Sinnott-Armstrong, Walter, "Consequentialism", The Stanford Encyclopedia of Philosophy (Winter 2023 Edition), Edward N. Zalta & Uri Nodelman (eds.), forthcoming URL = https://plato.stanford.edu/archives/win2023/entries/consequentialism/
Alexander, Larry and Michael Moore, "Deontological Ethics", The Stanford Encyclopedia of Philosophy (Winter 2021 Edition), Edward N. Zalta (ed.), URL = https://plato.stanford.edu/archives/win2021/entries/ethics-deontological/
GEUS. National boringsdatabase (Jupiter). De Nationale Geologiske Undersøgelser for Danmark og Grønland. https://www.geus.dk/produkter-ydelser-og-faciliteter/data-og-kort/national-boringsdatabase-jupiter
Rikke Markfoged, Consultant at Institute of Technology, Personal Communication, August 16, 2023.
Meegoda, J. N., Bezerra de Souza, B., Casarini, M. M., & Kewalramani, J. A. (2022). A Review of PFAS Destruction Technologies. Int J Environ Res Public Health, 19(24). https://doi.org/10.3390/ijerph192416397
Horst, J., McDonough, J. T., Ross, I., & Houtz, E. F. (2020). Understanding and Managing the Potential By‐Products of PFAS Destruction. Groundwater Monitoring & Remediation, 40.
Ivanova, I. (2023). Global brands lied about toxic "forever chemicals," new study claims. https://www.cbsnews.com/news/3m-dupont-pfas-forever-chemicals-hid-evidence-study/
Gaber, N., Bero, L., & Woodruff, T. J. (2023). The Devil they Knew: Chemical Documents Analysis of Industry Influence on PFAS Science. Annals of Global Health. https://doi.org/10.5334/aogh.4013
Vendl, C., Taylor, M. D., Bräunig, J., Gibson, M. J., Hesselson, D., Neely, G. G., Lagisz, M., & Nakagawa, S. (2021). Profiling research on PFAS in wildlife: Protocol of a systematic evidence map and bibliometric analysis. Ecological Solutions and Evidence, 2(4), e12106. https://doi.org/https://doi.org/10.1002/2688-8319.12106
GEUS. Status på aktuelle fund af PFAS I rå- og drikkevand. De Nationale Geologiske Undersøgelser for Danmark og Grønland. https://www.geus.dk/produkter-ydelser-og-faciliteter/data-og-kort/national-boringsdatabase-jupiter
Dennis Nielen de Raadt, studying BEng in Global Management and Manufacturing at the University of Southern Denmark, Personal Communication, October 9, 2023.
