Team:Groningen/Gold Human Practices

Integrated Human Practices


As described on our Human Practices page (and on our landing page and our project description page) the Dutch nitrogen crisis is a complicated problem, with various needs opposing each other (environmental, economical and food-security) and different actors who have very different interests. It is therefore important that anyone, including us, who tries to help find a solution to this problem, studies both the problem and any possible solution in both deepening and broadening research. Only when scientists step out of their ivory tower can they help develop a solution that not only addresses the actual problem, but is also responsible, safe and good for use “in real life”.

On our Human Practices page, we presented all the different things we did to research whether BYE-MONIA is responsible, safe and good for the world. On this Integrated Human Practices page, we present how we used the information from our Human Practices work to adjust our project so that BYE-MONIA is actually responsible, safe and good for the world. All leading to us proposing to use BYE-MONIA as described on our implementation page.

To ensure that we were able to adjust our project optimally after everything that we learned during our Human Practices work, it was needed to have a clear and concise summary of: the contribution of each activity; how it should lead/has led to adjustments in our project; and what our next steps should be afterwards. We have therefore, as described on our Human Practices page, already decided to process the results of our stakeholder interviews according to the AREA framework as brought to iGEM by iGEM Exeter 2019:

AREA framework that we used for our integrated Human Practices work. As brought to iGEM by iGEM Exeter 2019

We however also decided to process the results from our literature research and Human Practices tools according to this tool. For these two Human Practices activities, we will however skip the description step since this is explained on our Human Practices page. Moreover, for our Human Practices work we set up a Safe-by-Design (SbD) framework for our project. This framework was however mostly set up as a result of things we learned during our (integrated) Human Practices work, and is therefore already a take-away/integrated Human Practices on its own. We will therefore not process this SbD framework further, but decided to keep it on our Human Practices page as we think it does fit better on that page than on our Integrated Human Practices page. Lastly, we have executed several Human Practices collaborations, but since these did not change our project itself significantly, we have chosen not to mention them again on our Integrated Human Practices page.

All in all, by integrating our Human Practices work, we were able to determine the key values of our project and develop BYE-MONIA to uphold these.

How we integrated the take-aways from our Human Practices work to determine our key values that lead to our final project version of BYE-MONIA. Even though our Safe-by-Design work is officially our Human Practices work, in practice it became one of the key values of our project, hence the different colour of that building brick. Created with

Take-aways from literature research

From our literature review we learned the following about the background and scope of the problem we are trying to tackle: (excess) emissions of nitrogen-containing compounds, including ammonia, do indeed cause a problem in the Netherlands. When these compounds deposit in excess in nitrogen-sensitive areas, they harm biodiversity and the quality of habitats through eutrophication and acidification. In terms of deposition, more nitrogen is deposited in the form of ammonia than in the form of nitrogen oxides, in addition to which ammonia has a greater eutrophying and acidifying effect. All in all, the majority of Dutch nature areas exceeded limits for nitrogen in 2016. Excess nitrogen deposition can also have detrimental effects on Natura 2000 sites, which provide a safe haven for rare and threatened species, but are generally sensitive to nitrogen. Despite the fact that Dutch nitrogen emissions have decreased significantly since 1990, the decrease has stagnated around 2013 and it is still very uncertain whether the Netherlands will meet internationally agreed guidelines for maximum nitrogen emissions. Compared to other European countries, the Netherlands is also the second largest nitrogen emitter per square kilometer. In both the Netherlands and the EU, it is mainly agriculture that is responsible for nitrogen emissions, with, in the Netherlands, emissions from livestock playing a major role. A much-mentioned solution to the nitrogen problem is therefore to curb the Dutch livestock industry. However, this approach is not as easy as it might seem, because it could have adverse consequences for global food security, the Dutch GDP and social cohesion between farmers and government. For sources, please see our Human Practices page.

Our literature research mainly confirmed for us that with our project idea, decreasing ammonia emissions from cattle by using a feed additive, we were on the good track. Despite the fact that we recognized that the nitrogen problem is bigger than just ammonia emissions from cows, and that there are also other susceptible risks to Dutch nature besides excessive deposition of nitrogen, we knew for sure that the problem we wanted to tackle (excess ammonia deposition) was actually a problem and that we would address the biggest cause of this problem (emissions from livestock/cows).

We did however decide to be very open everywhere on our wiki on that our project would, on its own, not be able to fix or even address the entire problem of the Dutch nitrogen as the entire problem for Dutch nature is bigger than just the deposition of ammonia. Although this may result in a less compelling story, we felt it was important to also clearly show the pitfalls and shortcomings of our project as we do not want to create unreasonable expectations/hope about our project, especially in a discussion that already affects so many people (emotionally and financially).

Finally, we decided to keep the story of our project centered around the situation in the Netherlands as much as possible, since our research showed that the Netherlands is indeed - relatively - one of the top producers of nitrogen emissions in Europe. We did however decide that while developing our project, we would take into account as much as possible that our project could be used anywhere in the world, even in areas where the agro-technological or financial situation was perhaps less favorable than in the Netherlands. Moreover, we also realized that using our project in the Netherlands also has beneficial effects for other countries. Because if our project reduces Dutch ammonia emissions, it also reduces Dutch ammonia emissions abroad: after all, ammonia emissions do not stop at the border.

After our in-depth literature review, we decided to:

  • Continue with our current project idea where we are trying to counteract (part of) the nitrogen crisis by reducing ammonia emissions from cows.
  • Emphasize very clearly on our wiki that we are only addressing one part of the Dutch nitrogen crisis (ammonia emissions) but that there is another part as well (nitrogen oxides).
  • Focus the story of our project mainly on nitrogen emissions and deposition in the Netherlands specifically.
  • Engage in further research (in the form of Human Practices tools and Stakeholder interviews) to learn even more about the context of the Dutch nitrogen crisis.

Take-aways from Human Practices tools

Stakeholder analysis

Our stakeholder analysis mainly gave us insights into how the different stakeholder groups are connected to our project and in what would be the best way to involve these different stakeholder groups in our project. For example, we learned that it would be best to “actively engage” several governmental stakeholders, farmers and people who are tied to nature (ecologists and climate change NGOs). Moreover, we learned that it would be better to keep the people who are concerned (about nature, about GMOs in general and about the safety of our project) informed about our project. In addition, as already mentioned on our Human Practices page, it stood out to us that especially the government can have a lot of influence on our project, sometimes even more than possible owners of the final project. Lastly, it is also striking that, besides livestock, especially nature (both animals and plants) and future generations have little influence on the problem and the project, while they are precisely the groups that are (or could be) most disadvantaged by the nitrogen crisis.

After completing our stakeholder analysis, we decided to mainly reach out to farmers, ecologists, governmental organisations and organizations involved in measuring nitrogen emissions and depositions (such as the RIVM), since they are mainly represented in the “actively engage” quadrant. Moreover, since we noticed that many stakeholders in the “keep informed” quadrant would mainly be interested in the risks of our project and how we mitigate those, we adjusted our project so that safety would become a bigger priority. For example, we decided to use the WAIR-tool, which has an extensive section on safety, and we decided to reach out to stakeholders and attend meetups that could help us with designing our project in an Safe-by-Design way to mitigate any risks our project might pose.

After our stakeholder-analysis we decided to:

  • Start up our stakeholder interviews in which we would reach out to farmers, ecologists, the government and scientists who measure nitrogen emissions and depositions
  • Make safety one of the key values of our project, by doing more background research on possible safety issues and dual use potential of our project and by setting up our own Safe-by-Design framework for our project
  • Reach out to stakeholders/attend meetups that are specifically catered to Safe-by-Design and the prevention of dual use
  • Fill in the safety-section of the WAIR tool to be able to investigate any safety risks our project might pose and how we can mitigate those risks

WAIR tool

Our WAIR-tool gave us insights into: the stakeholders that are involved in our project; possible future perspectives that can influence our project; the needs and problems our project addresses; the safety of our project; and the alternatives to our project.

In terms of the stakeholders involved in our project, we learned that there could be controversies between the different stakeholder groups (as is actually also seen “in real life”). Controversy between groups could center around: if economy should trump nature or if nature should trump economy; if the other party is performing their work correctly; if the other party has reasonable expectations; or if genetic modification is an desirable approach to take to solving problems. All in all, most of the conflicts seem to be due to stakeholders not feeling taken seriously.

In terms of future trends and developments, we learned that both a change in dietary habits and a change in the status of the nature areas could impact the impact of and need for our project. We learned that even the ‘most ideal’ futures have their downsides and even the ‘most negative’ scenarios have their upsides. For example, sometimes technological advancements are only created once the situation has escalated. However, it is difficult to imagine a future based on only a few changing parameters. The future is complex and making an accurate prediction is close to impossible.

In terms of needs and problems our project addresses, we found out that we are actually working on quite a diverse set of Sustainable Development Goals (SDG’s). Not only does BYE-MONIA address environmentally-related SDG’s, for example #14 and #15, but we also address more economically related SDG’s, for example #1 and #8. Lastly, BYE-MONIA is very involved in (food)-production related SDG’s such as #2 and #12. However, if our project is not designed or executed responsibly and safely, our project might actually be able to cause harm to the environment or income equality. We identified 3 main risks of our project: possibility of greenhouse gas emissions; possible danger of alpha-amylase to nature and plants; and the possibility that our project heightens economic inequality. However, we also saw that much of these problems could be avoided or mitigated by doing enough background research and applying fitting Safe-by-Design decisions thereafter.

A list of all the Sustainable Development Goals our project addresses

In terms of safety and SbD, the WAIR-tool also helped us look more extensively at these issues as well. By first imagining what events could occur and what risks these events might pose, we afterwards were able to define how these risks could possibly be mitigated and what we could design to help mitigate these risks. The main things we took from this exercise is: that we should pick/design a safe enzyme; that we should pick a safe chassis; that engineering a kill switch would greatly add to the safety of our system in the industrial setting; that we should promote environmentally friendly choices when it comes to our project, especially when it comes to heating up the MOF; we should strive towards maximal production and minimal resources to make sure our project is also accessible for less wealthy farmers; that safety testing on similar feed additives could teach us a lot about the safety of ours; and that our Human Practices work should be as open and honest as possible to prevent any false hope or promises arising from our project.

Lastly, the WAIR tool helped us to look at alternative solutions for the nitrogen crisis. More specifically, we looked at how well these alternatives work towards solving the environmental and economical problems that are tied to the nitrogen crisis. We quickly learned that there was a pattern that if any solution for this multi-level problem focussed mainly on one element of the problem, that the solution was most of the time at risk of being rather unethical. We also learned that there are plenty of alternatives for one or two of the problems we want to address, but none of them are able to help solve all of the problems at the same time. However, our main conclusion was that competing against other solutions to the nitrogen crisis will not be beneficial to any of the problems we are trying to tackle. The best solution to the problem would therefore be a combination of different solutions that can work together to help tackle the different problems caused by the nitrogen crisis.

After completing the WAIR tool, we decided to take elements from each section to adjust our project.

From the stakeholder section, just as from our stakeholder analysis, we decided to mainly focus on reaching out to farmers, ecologists, governmental organizations and organizations that measure nitrogen emissions and depositions. Also, just as from our stakeholder analysis, we decided to make safety a bigger priority. Moreover, we decided to use our stakeholder interviews, specifically those with farmers, to find out how we can make a solution workable in practice, instead of just in theory. Lastly, seeing that we aim to communicate any risks of our project honestly and clearly, and since we ourselves are also involved in the debate between strain engineers and people who are hesitant against GMOs, we asked our education team to dedicate one of their comics to the risk of synthetic biology if adequate safety measures are not put in place. Our education team was excited about this idea, but in the end did not have the time to work on it.

From the futures section, we were not able to make any major adjustments to our project. Mainly because, as stated before, it is difficult to imagine a future based on only a few changing parameters. The section did however help us understand the durability of our project better and how its durability might change if certain parameters change. Moreover, it helped us understand why some problems are solved so late: because often real investment in technology, research and a new (appropriate) solution is only made when the situation is really dire.

From the needs and problems section, we learned that our project tackles a diverse set of goals, from food security to financial security to environment. This was a great finding since our idea from the start was to address the multiple diverse aspects of the nitrogen crisis with our project. We therefore decided to continue promoting our project in such a way. However, more importantly, we decided to actively work on employing Safe-by-Design principles and set up a Safe-by-Design framework for our project to minimize the risk of BYE-MONIA causing greenhouse gas emissions, adverse effects to nature and plants or economic inequality. More specifically, in our Safe-by-Design framework we decided to promote sustainable choices (such as sustainable energy sources to heat up the MOF), pick a safe enzyme and strive for maximal production of alpha-amylase with minimal resources. We had also hoped to be able to interview a stakeholder who is involved in plant-sciences and could therefore tell us more about the potential adverse effect of active alpha-amylase ending up in nature. However, due to time constraints this was not possible within the timespan of iGEM.

The safety section had a major impact on our project, most notably on our Safe-by-Design framework. Even though other sections of the WAIR-tool and our stakeholder interviews helped us see the need for employing a Safe-by-Design framework, it was mostly this section that helped us make a clear overview of the different Safe-by-Design decisions we wanted to make in our Safe-by-Design framework. The main decisions we made were: picking/designing a safe enzyme and host organism; designing a kill switch; promoting environmentally friendly choices; and designing our project so that it ensures maximal production with minimal resources. Apart from these Safe-by-Design choices this safety section also inspired us to look at safety testing done on similar feed additives and to communicate our Human Practices work as open and honest as possible, in which mentioning the pitfalls, risks and limits would not be avoided.

Lastly, after filling in the alternatives section we learned that not many other projects address all aspects of the nitrogen crisis that we are addressing, but that other projects bring certain benefits that our project does not. Therefore, our main adjustment was not to use our project in isolation, but to use it in conjunction with other solutions to the nitrogen crisis. This also coincided nicely with what we had learned during our literature review. We decided to communicate this clearly on our wiki and in the story of our project.

Much of what we learned from each specific section of the WAIR tool coincided with what we learned in other sections of the WAIR tool (and what we learned from our other Human Practices activities). Therefore, we no longer broke down the follow-up steps by WAIR tool section but rather combined them into an overall roadmap. Our follow-up steps after completing the WAIR tool are to:

  • Reach out to farmers, ecologists, governmental organizations, scientists who measure/model ammonia emissions and depositions and scientists involved in Safe-by-Design for stakeholder interviews
  • Dedicate our stakeholder interviews, specifically those with farmers, to find out how to make our project workable in practice
  • (If we had had the time) Dedicate one of our comics to the safety risks that are important in synthetic biology
  • Promote that our project tackles a diverse set of (Sustainable Development) goals surrounding economics, the environment and food-security
  • Make safety one of the key values of our project, by doing more background research and dedicating stakeholder interviews on possible safety issues and dual use potential of our project
  • Set up our own Safe-by-Design framework for our project in which: promoting sustainable choices; picking a safe organism; picking a safe enzyme; designing a kill switch; and maximal production and minimal resources are for sure included as Safe-by-Design decisions
  • Look at safety tests performed on similar feed additives to learn more about their possible safety issues and how to mitigate those in our own project
  • Be open and honest about the potential, risks, pitfalls and limitations of our project in all the communication surrounding our project
  • Communicate clearly, already on the landing page of our wiki but also in all the communication surround our project, that our project will not be the sole solution to the nitrogen crisis in the Netherlands and should be used in combination with other (non synthetic biology based) solutions to ensure optimal results

Take-aways from stakeholder interviews and meetups

Whilst doing our literature background research and working on our Human Practices tools, we also spoke to several stakeholders, whose input significantly shappend our project. Moreover, during our iGEM session we attended several meetups that helped us shape our project. In fact, with the input we got from these interviews and meetups, we were able to further deepen our background research and better fill out the Human Practices tools. We reached out to the following groups:


As indicated on our Human Practices page, we chose quality over quantity for the stakeholder interviews and it took some time to set up our stakeholder framework that maintained data protection and informed consent. As a result, we were unable to conduct as many interviews as we would have ideally liked. However, due to the great preparation prior to these interviews, which included us preparing the questions in advance and the use of the AREA framework to create an extensive summary with concrete follow-up steps after each interview, we were able to make the most out of every interview we had.

Wondering how these interviews and meetups led to tangible changes in our project? Below we included a timeline of who we reached out to and what they taught us about: the impact of the nitrogen crisis on nature; the role of agriculture in the nitrogen crisis; measuring and modeling ammonia emissions, Safe-by-Design and (preventing) Dual-Use Potential.


Name stakeholder: Biosecurity and dual use webinar organised by Bureau Biosecurity of the RIVM

Position of stakeholder within company: The RIVM is the Dutch national Institute for Public Health and the Environment


Date of meeting: 21-6-2021


RIVM's Bureau Biosecurity organized a series of webinars in 2020 and 2021 to raise awareness among researchers about biosecurity and dual-use risks. This webinar was the third in a series of three and was most relevant to us because it was specifically targeted at researchers and laboratory staff. Therefore, the goal of this webinar was to increase awareness of biosecurity and dual-use risks among this target group. The webinar tackled questions such as: “Can the results of your well-intended research be misused? Would you notice if some of your Eppendorf tubes disappeared from the freezer and how trustworthy are your colleagues? Can your research be a fundament for bioterrorism?”. The first part of the webinar covered different aspects of biosecurity including practical examples and solutions to everyday biosecurity issues participants can face in their research project. The second part of the webinar covered potential dual-use aspects of research. Also two keynote speakers were present:

  • Piers Millett, the vice president for Safety and Security at iGEM. Piers spoke about examples of Biosecurity Challenges from iGEM.
  • Daniël Schepers, a Policy Officer for Export Control and Strategic Goods at the Ministry of Foreign Affairs. Daniël spoke about the export control of dual-use goods.


During the webinar we got a lot of information about the difference between biosecurity (protecting the world against the bug) and biosafety (protecting the bug against the world). We already knew some of it, but it was nice to have a refresher and make sure we were up to speed since now, for the first time for most of us, we would set up our own project and run our own lab. The webinar dealt with the most important rules for biosecurity such as leaving the doors of the lab locked; locking and organizing fridges in the lab; and never letting visitors of the lab unsupervised. All in all these rules helped us ensure that we worked on our project in a safe manner.

The webinar also offered us additional resources in case we needed more information on biosafety. They let us know that we could reach out to the Biological Safety Officer of our own University/faculty, that we could go through the WHO laboratory safety manual or that we could use a site created by Bureau Biosecurity1.

For the next section Piers Millett spoke about Biosecurity Challenges that previous iGEM teams encountered or were trying to tackle. We learned that when ordering DNA from gene synthesizing companies, that these DNA fragments are normally checked to see if they contain any harmful sequences or sequences that can be used for malicious intentions.

The next section of the webinar dealt with what dual use potential was and what export control rules are in place to try and prevent the export of dual use items. We learned that even the best-intended research project can have dual use potential. Moreover, we learned that virtually all projects have dual use potential if people are creative enough. Therefore export control is in place for not only tangible goods (such as vials containing GMOs) but also for the sharing of information to foreign countries. We learned that as soon as you ship tangible products or collaborate internationally that you might need an export license. All the information on how to get such an export license can be found on the website of Bureau Biosecurity2, which is a part of the Ministry of Foreign Affairs in the Netherlands.


Even though our team already knew some of the things that were explained during the webinar, we did get a good refresher of the safety procedures we did know about (and why they are important). Moreover we learned more about the difference between biosafety and biosecurity and the importance thereof. Therefore, this webinar mainly helped us to set up and confirm our rules in our lab to ensure the biosafety and biosecurity of our project. We therefore decided to keep our lab firmly locked, keep our fridge very organized and always accompany any visitors to our lab. Moreover, after learning what dual-use potential is and that virtually every project has it, we decided to look into the dual use potential of our project and how we could mitigate this. In addition, because at that time we were discussing with our partner team Tirupati whether we could share Saccharomyces strains with each other, this webinar taught us that there are different export rules and forms that we need to take into account (also timing wise) before sending the strains. In the end, we did not send strains to Tirupati, but had we done so, after this webinar we would definitely have followed the rules and forms that were explained during the webinar.

Our next steps:

  • Set up new/additional lab rules to ensure biosafety and biosecurity even more
  • Share the new lab rules with the rest of the team that did not attend the webinar
  • Learn more about the dual use potential of our project and how to prevent this by reaching out to stakeholders involved in Safe-by-Design and filling in brainstorming tools

Name stakeholder: Leandros Tsiotos

Position of stakeholder within company: iGEM ambassador to Europe


Date of meeting: 23-6-2021


Our team decided to partake in the Values and Risks workshop offered by iGEM. This values and risks workshop was intended to help teams start up their Human Practices approach by taking the teams through a step-by-step journey of an exemplary team that made some unfortunate choices regarding Human Practices on the way. This way, the team that attend the meetup would learn from the examples and prevent these faults in their own project. The workshop is interactive with the participants answering various questions. Unfortunately, not enough people showed up to have a proper discussion on the original date. However, Leandros was kind enough to give a private workshop for our team only! As we saw potential added benefits of this workshop, we got together with a few additional team members and went through the programme designed by Leandros.

During the workshop we discussed at which points during the design process of our project, safety considerations should be taken into account. Leandros also went into more details about the organisation behind iGEM and which committees might be relevant for our project.


We went into the workshop fairly well prepared. During the Biosecurity workshop given by the RIVM, we had already learned quite a bit of potential hazards when doing research in the field of microbiology. Furthermore, we had already started our own research on how to conduct proper Human Practices, by looking at what old iGEM teams did. Therefore, we were able to identify most of the safety issues discussed during the workshop. The workshop was valuable nonetheless as it reaffirmed our beliefs that safety is important during various steps of your project development: not only is safety in the lab important, but also safety during stakeholder interviews (in the sense of informed consent and keeping ethics in mind).. Moreover, it helped us reaffirm that we were on the right track with setting up our own Human Practices approach with data-security and informed consent in mind.

After the workshop, Leandros also showed which committees within iGEM we can contact if we were to have any questions regarding iGEM’s rules of conduct (, Human Practices (, Safety & Security ( or Diversity & Inclusion ( The potential awards given by these committees were also brought to our attention.


While safety was already one of our main priorities, we decided to do even more research on how to conduct safe research and re-evaluate our project with respect to potential safety hazards, especially those that we might encounter in future stages of our project. For this, we among others continued reading through the Dutch General Data Protection Regulation and looking into if we needed to do legally anything else data-privacy-wise (which we already started doing before the workshop).

Afterwards, we had a discussion on whether we would like to commit to contending for one or more of the awards. Given the time and effort we already put in our Human Practices and the Safety and Security of our project, we figured we could give competing for those awards a shot.

Our next steps:

  • Do more research on what iGEM teams previously nominated for the Integrated Human Practices and Safety and Security awards did and what we can learn from them.
  • Re-analyse our project and identify potential hazards.
  • Continue reading through the Dutch General Data Protection Regulation and see if there is other Dutch data-privacy legislation that applies to us.


Name stakeholder: all Dutch iGEM teams (Leiden, Delft, Maastricht, Eindhoven and Wageningen)

Position of stakeholder within company: Student members of each respective iGEM team


Date of meeting: 02-07-2021

Description: This meetup was organised by iGEM Wageningen so all Dutch iGEM teams of 2021 could get to know one another and present their project. Moreover, some keynote speakers were present to give a presentation about temporal and spatial patterns in (synthetic) biology and about the dynamics of proteins. As the final part of the meetup, teams were paired up for a short Human Practices workshop. During this workshop each team asked the other team questions about potential safety/application issues in each other's project and the teams offered each other tips afterwards. At the end of the Human Practices workshop, the teams had to shortly present to all other teams what they learned from the Human Practices workshop. From this workshop we learned several things that contributed to the development of our project.

Contribution: During the Human Practices workshop, we were paired up with members of iGEM team Wageningen 2021. They asked us whether we already knew if the feed additive we planned on producing was safe for the cows itself: whether this feed additive could possibly have any adverse effects on the health of the cows. We learned that even though we would not expect our feed additive to have any adverse effects on the health of cows, it would still be worthwhile to look through literature to find any evidence to either confirm or deny this suspicion. Moreover, team Wageningen advised us to look into how we could make our end-product appealing to farmers: how can we make sure that the farmers trust the safety of our project and actually want to use it?

Adjustments: After the Dutch iGEM meetup, we decided to spend more attention on the safety part of our project. Even though we already suspect that the feed additive would be safe to cattle, we want to ensure that this suspicion is correct. Moreover, we want to look into how we can ensure that our project is actually appealing to farmers. In order to do this, we first want to look into the wants and needs of farmers and how our project could possibly play into that. Once we have a clear image of what it is farmers need, and we are sure that we have designed our project to be safe, we can work on incorporating these new insights into our pitch about the project.

Our next steps: Since we would like to confirm that our feed additive is in fact safe for cows, we will search for an expert on GMO safety to interview them about the risks of heterologous protein expression and synthetic biology in the food industry. Moreover, we will go through literature to look for similar feed additives and their effects on the health of the organisms they are being fed to. We hope to learn from their findings and adjust our project accordingly afterwards. Lastly, we will reach out to farmers to interview them about their wants and needs; how we can gather our project to suit their wants and needs better and how we can ensure that our project is and feels safe to them as well.

Name stakeholder: Other iGEM teams that work with or are interested in biodiversity (UNSW Australia, Stony Brook USA, USYD Australia, NU Kazakhstan)

Position of stakeholder within company: Student members of each respective iGEM team


Date of meeting: 15-07-2021

Description: iGEM team UNSW Australia 2021 invited us and other iGEM teams who dealt with biodiversity to their Biodiversity Symposium. The event was a great way to get to know other iGEM teams; learn how other iGEM teams tackled similar and different biodiversity issues and to learn more about biodiversity in general. During the symposium we listened to the presentations of the other iGEM teams about their projects and we presented our project, after which we got encouragement and feedback from the other teams. The feedback from the other teams on our project also contributed to the development of our project.

Contribution: After our project presentation, there was room for the other teams to ask questions and give feedback. We learned that the other teams were very excited about how we used synbio to create a win-win situation for multiple communities and demographics and that we related our project to the local community around us. Moreover, similarly to the Dutch meetup, the other teams questioned if we were sure that our feed additive was safe for cattle. Even though we are already looking into literature and trying to contact stakeholders to learn more about the safety of our feed additive, these questions were a welcome confirmation that we are indeed allocating our time to find worthwhile answers to important unknowns about our project. Lastly, the other iGEM teams advised us to look more into ‘the numbers’ of our project. They advised us to look into how much the ammonia/nitrogen emissions in our country should be reduced; by how much our project could reduce these nitrogen emissions and how much the current solutions to the nitrogen crisis costs and how that compares to the costs of implementing our system nationwide.

Adjustments: We learned a lot from the biodiversity meetup about how we could adjust our project to meet the needs of our stakeholders even better. First of all, even though not an actual adjustment, we decided to keep incorporating the benefit of our project to both farmers and the environment in our project pitch. This way we can clearly demonstrate why our project is an innovative solution to a local problem since it focuses on the needs of multiple stakeholders. Moreover, we decided to continue with our efforts to learn more about the safety of our and similar feed additives to both cows and the environment. Lastly, we decided to investigate more how big the nitrogen crisis in the Netherlands is; how much our project could help with fighting the nitrogen crisis and what the financial costs of our project would be compared to the financial costs of the current solutions.

Our next steps: Since the dual benefit (to both farmers and environment) of our project is what makes our project so interesting and innovative, we decided to keep incorporating this point in our project presentation. Moreover, we will continue reading literature and reaching out to stakeholders who know more about the potential adverse effects of (similar) feed additives to cows. Lastly, we will reach out to stakeholders we are involved with measuring nitrogen/ammonia emissions and ask them how high the nitrogen/ammonia emissions in the Netherlands are; how much the current solutions are helping in decreasing the ammonia/nitrogen emissions; by how much the emissions need to drop and what the best sources are to find accurate numbers on nitrogen/ammonia emissions ourselves. We will do all of this in order to assess how big the problem is; by how much our project could help solve the problem and how financially feasible our project would be in comparison to current solutions to the nitrogen crisis.


Name stakeholder: Cécile van der Vlugt-Bergmans and Marja Agterberg

Position of stakeholder within company: Both Cécile and Marja work at the RIVM (Dutch institute for Public Health and the Environment). Cécile is a senior risk assessor of GMO’s and Marja is a senior risk assessor GMOs and project manager at Bureau GGO


Date of meeting: 02-08-2021

Description: Just as previous years, the RIVM reached out to Dutch iGEM teams to collaborate with them on the safety-aspects of their project. Cécile offered our team the opportunity to have an interview with RIVM experts to help us think about incorporating Safe-by-Design (SbD) in our iGEM project and to discuss all kinds of safety issues related to our project. We were able to choose whether we preferred to have one longer interview, or two shorter interviews. We decided to go for the two shorter interviews, so we could discuss the progress we made since the first interview during the second interview.

This AREA framework is dedicated to the first interview. During this first interview, we spoke with Cécile, who has been working on SbD and with iGEM teams for multiple years and with Marja, who works with GMO’s in contained use and is responsible for reviewing the applications for several safety certificates.

Lastly, we were given a SbD-case to discuss amongst our team before the interview to learn what taking a SbD approach can entail. The second interview and the outcomes of the SbD case will be discussed in the AREA-framework “RIVM Safe-by-Design, meeting #2”.

Contribution: During the meetup we first told Cécile and Marja about our project: what the problem was we are trying to tackle, what the scope and the effect of the problem is and how we were trying to tackle the problem. Afterwards, Cécile and Marja questioned us about several safety aspects of our project.

First of all, Cécile asked us about our working practices in the lab and if we adhered to standard microbial safety practices. We informed Cécile that we did and that all of our team members who entered the lab had obtained their certificate of Safe-Microbial-Techniques and that we were in close contact with the biological safety officer of our faculty.

Moreover, Cécile wondered whether we knew for sure that our feed additive is safe for cattle and humans. We mentioned that, based on the mechanism of action, we expected that it would be. However, we did not have any proof of this suspicion and were still looking into it. Cécile therefore gave us the tip to look at the scientific and safety review of similar feed additives as performed by the European Food Safety Authority (EFSA).

In addition we spoke with Marja and Cécile about the production of our feed additive and the safety regulations concerning such production processes. We mentioned that we were still unsure whether we wanted to produce the feed additive ourselves in a lab/industrial setting and then sell this feed additive or that we wanted to develop a kit that farmers could use to produce their own feed additive with our GMO; therefore giving the farmers more autonomy. Marja and Cécile informed us that if we were to produce such a kit; that that would substantially complicate the process considering that any work with a GMO should be done according to strict safety standards, which not every farm would have the capacity for. For example, a farmer would need a biological safety officer, permission for a BSL-1 activity and a WABO-certificate and all the work needs to be done in a ML-1 laboratory. Alternatively, farmers could work together to set up a shared lab and produce alpha-amylase together, but it remains questionable whether every farmer would have the capacity and time for this as well. Marja and Cécile therefore mentioned that a way out of this problem would be to get our GMO extensively tested for safety and get it market approved. However, this would be a long trajectory with lots of regulations and public opinions to overcome. And there is little experience with getting GMO’s market approved in the Netherlands.

Adjustments: While not a definitive adjustment as of now, we decided to look through papers of the EFSA to find more information on the safety of similar feed additives. Based on this information, we could then decide to change the design of our project in case similar feed additives face any safety issues.

The most important adjustment we made after this interview was that we realised that the possibility of developing a kit, that farmers could use on their own, would be rather infeasible and possibly unsafe in practice. We therefore decided to explore other options: both producing the feed additive on our own in an -by safety measures regulated- industrial setting as well as the possibility of setting up a shared lab for farmers to use.

Our next steps: Before the next interview with Cécile, we would go through a review of the EFSA on a similar feed additive[1] to investigate any safety concerts of the use of alpha-amylase as a feed additive for cattle and adjust our project accordingly. Moreover, we will explore the possibility of producing our feed additive in an industrial setting and in a lab shared by farmers. For the latter option specifically, we will also try to reach out to farmers to find out more about their opinions on the feasibility and desirability of this option.


  1. European Food Safety Authority. 2013. “Scientific Opinion on the Efficacy of Ronozyme® Rumistar (Alpha-Amylase) as a Feed Additive for Dairy Cows.” EFSA Journal 11(10).

Name stakeholder: Anton Stokman

Position of stakeholder within company: Owner of a dairy farm


Date of meeting: Monday 02-08-2021

Description: Anton Stokman is the owner of the Stokman Koudum, a sustainable, GMO-free and animal-friendly farm in the Netherlands. In his farm, Anton is continuously looking for new ways to further develop the sustainable way of keeping dairy cattle, as a test farm for new technological projects. He also developed the Free Choice Concept, by which cattle are allowed to leave the barn to the farm and come back whenever they want to. His knowledge and expertise on farming and receptiveness to the new technologies, encouraged us to get into contact with him. This meeting gave us the opportunity to gain a better understanding and wider picture of a cattle farm in the environment.

Contribution: During the meeting we asked Anton about his experience with the nitrogen crisis and his opinion on the problem and current solutions. We learned that the main effect of the nitrogen crisis on farmers is that it is hard to expand their business, and that current regulations in the Netherlands are very strict and sometimes hard to resonate with for farmers. We also asked Anton about the amount of nitrogen/ammonia emissions in his barn and what measures he currently takes to fight those. Anton let us know that since most modern dairy barns are highly ventilated, ammonia is rather equally distributed inside the barn. Moreover, Anton provided us with the solutions he has employed in his farm to reduce the nitrogen emissions, such as injecting manure into the soil; separating manure from urine and adding water to the manure. In the end, we asked for his idea about our project and how we could improve it. In the first place we came to know that a feed additive that is regulated by the government is preferred to a kit which requires the farmer’s effort and financial investment to set up. He advised us to introduce our project as a solution which helps with animal welfare and optimal dairy production rather than focusing on the nitrogen crisis, when approaching the farmers. We also learned that GMO-free farmers would most likely only become interested in using our GMO-based project in case the aim of this use is well defined and if the (financial) benefits outweigh the costs.

Adjustments: We decided to slightly change our way of approaching and explaining our solution to the nitrogen crisis to farmers. Rather than focussing on the negative effects on nature our project helps prevent, we will focus more on the positive effects our project has on the milk production and welfare of cows. Also, as the ammonia concentration should be high for the MOF, whereas ammonia concentrations in modern cattle barns are rather evenly distributed throughout the farm, we have to do more research on where to place the MOF in the barn, so that it works properly. For further implementation of our current solution in the farms we need to produce the feed additive in a cost-effective manner, so that farmers see it as a profitable addition that increases the farm’s annual revenue. We therefore decided to not design a kit for farmers to use, but would instead focus on central production of the feed additive which can therefore be produced more cost-effectively.

Our next steps: We now aim to contact more farmers to see their opinion on our project and see if they will be willing to use GMO’s as feed additive in their farm. Moreover, we aim to contact more experts in the field of measuring and modelling ammonia emissions from livestock as to get a better idea of where to place the MOF.

Position of stakeholder within company: Scientific staff member agricultural emissions at the RIVM (Dutch National Institute for Public Health and the Environment)

Date of meeting: 9-8-2021

Description: This stakeholder has extensive knowledge of how nitrogen emissions of the Dutch agriculture are modeled and also works with this professionally. Our team therefore set up a meeting with this stakeholder in order to gain new insights on the main sources of nitrogen emissions in the Dutch agricultural sector, other sources of nitrogen emissions that end up in the Netherlands and how our product could help reduce these emissions.

Contribution: The stakeholder pointed us to the website and showed us how to find and compare the emissions of various categories of livestock, farm houses and manure dispersion techniques. also shows how the overall nitrogen emissions of the agricultural sector compares to the nitrogen emissions from the transport and building sectors. We also learned that the website from Eurostat could be used to find data on nitrogen emissions in other European countries and compare this with the data about Dutch nitrogen emissions.

Furthermore, the stakeholder told us which measures to reduce ammonia emissions in the agricultural sector are already put in place. While these measures have caused a substantial decline in emissions during the period of 1990 to 2010, the last decade we have hit a plateau and emissions are no longer in remission. This initial decline can be mainly attributed to the decrease of protein in the diet of livestock. In recent years, in fact, they have been steering toward optimizing protein content of the feed with the help of feed consultants. We have currently reached the point where feeding the livestock even less protein could result in the livestock not having enough buffers in case of an illness. Therefore a further reduction of the protein content in the feed of livestock should be done in collaboration with a feed consultant. However, our stakeholder indicated that recent research from the WUR has shown that some optimization is still possible.

Two other methods which are currently being investigated in order to reduce emissions by dairy livestock in particular are (1) Increasing the hours the animals spend in a pasture and (2) diluting the manure with water before fertilising the soil with said manure. These two measures, combined with the reduction of protein content in the feed, are predicted to yield a reduction of 9% to 10% of the total nitrogen emissions in the dairy sector[1].

While this reduction is substantial, it is not enough to reduce the emissions sufficiently enough to go under the critical limits set for certain nature reserves. Even if we were able to reduce the dutch nitrogen emissions to 0, these critical limits would still be exceeded as approximately 1⁄3 of all nitrogen deposited in the Netherlands originates from our neighbouring countries[2]. This highlights the fact that the reduction of nitrogen emissions should be an international effort.

After explaining our project, the stakeholder made a few useful comments:

  1. As we are planning on using alpha-amylase in a similar manner as DSM has been doing for some time, there could be patents preventing us from making it a commercial project.
  2. The Metallic Organic Framework (MOF) we plan on using for filtering the air from ammonia seems to work in a similar manner as microbial based “air washers”. These air washers could offer us insights in the best place to place our MOF in the farm.
  3. Even though we market our product as not harmful for the environment, we do plan on heating up the MOF to 200 degrees. For this a lot of energy will be needed, which could be produced in an unsustainable manner and therefore lead to the emission of harmful greenhouse gases.

As for the main selling point for our product, the stakeholder remarked that our product is both beneficial to the farmers -in the sense that the cows would be able to produce more milk with the same amount of feed- while also reducing nitrogen emissions, creating a win-win situation.

Adjustments: Based on the interview with the stakeholder we decided to adjust our project in the following manner:

  • We would focus on using sustainable energy sources to heat up the MOF, as to avoid the release of greenhouse gases as much as possible.
  • While no definite adjustments as of yet, we decided to explore the science behind an microbial “air washer” further. Not only to get more information on the best place for the MOF but also to look into alternatives for the MOF. A microbial “air washer” could possibly be a less energy dependent alternative to our current implementation of the MOF while still extracting the ammonia needed for the production of our alpha-amylase.
  • While no definite adjustments as of yet, we decided to make use of the websites and Eurostat. We will use those websites to get more data on the different sources of ammonia emissions in order to locate the main sources of ammonia emissions. This will allow us to more effectively design our project to make the most impact.

Our next steps:

  • Delve into the website We can use this website to identify where exactly the Dutch nitrogen emissions originate from and how much of a reduction could theoretically be obtained through the usage of our product.
  • Delve into the website from Eurostat. We can use this website to identify non-Dutch sources nitrogen emissions and compare those to the Dutch nitrogen emissions
  • Do more research on the subject of how “air washers” can be used in barns[3]. This could give us more information on the optimal placement of the MOF or it could even be a more sustainable alternative to the MOF such as the designs of the Lely Sphere.
  • Follow-up on the potential usage of a potentially patented product.

Finally, we will also read up on the current government policy and regulations regarding nitrogen emissions listed in the sources sent by the stakeholder[4],[5].


  1. Rijksoverheid. “Advies ‘Doorrekening Bronmaatregelen Stikstof in de Melkveehouderij’ | Kamerstuk | Rijksoverheid.Nl.” (August 21, 2021)
  2. Rijksinstituut voor Volksgezondheid en Milieu (RIVM). “Vragen En Antwoorden over Stikstof En Ammoniak | RIVM.” (August 21, 2021).
  3. Rijkswaterstaat Ministerie van Infrastructuur en Waterstaat. “Rav En Huisvestingssystemen - Kenniscentrum InfoMil.” (August 21, 2021).
  4. Mosquera, J et al. 2021. “Monitoring van Methaan-, Ammoniak-, En Lachgasemissies Uit Melkveestallen Praktijkmetingen in de Periode Oktober 2018-Februari 2020.” Wageningen university & research.
  5. Vink, Martijn et al. 2021. “NAAR EEN UITWEG UIT DE STIKSTOFCRISIS.” Planbureau voor de Leefomgeving.

Name stakeholder: Carsten Schep

Position of stakeholder within company: Researcher at Wageningen Livestock Research; sub branch of Livestock and Environment Research with a focus on emissions


Date of meeting: Thursday 26-08-2021

Description: Carsten grew up on a farm in the Netherlands and studied Agrotechnology with a research on pit emissions at Wageningen University. As such, Carsten has both good practical and theoretical knowledge on how and where ammonia is produced on farms; the effect of measures to combat the part of the nitrogen crisis caused by livestock barns and working on farms in general. During his study Agrotechnology, Carsten was trained in multiple disciplines of agro technique in order to be “the spill in the web” that can combine insights from modeling to plant sciences. Moreover, Carsten followed multiple courses and executed study projects on ammonia emissions from farms. After his study, Carsten started working at the Research branch of Wageningen University. For his profession, Carsten drafts and executes research proposals from companies/farms/institutions about emissions coming from barns. As such, this meeting with Carsten helped us gain a deeper understanding of how cattle farms are involved within the nitrogen crisis and if our solution would be feasible in practice.

Contribution: During the interview we first spoke with Carsten about how (nitrogen) emissions are measured and what the best sources are to find numbers about ammonia emissions from farms. We learned, just as with a previous stakeholder interview, that Emission Registration is indeed the best source to find accurate numbers about ammonia emissions. Moreover, we learned that when speaking about the nitrogen crisis, people most often refer to nitrogen emissions numbers instead of nitrogen deposition numbers since it is rather impossible to measure deposition accurately. To do so, one would have to use traceable molecules, which would be hard to do in practice. Therefore, depositions numbers are often the results from models instead of only measurements. Furthermore, Carsten explained to us that after a certain altitude, nitrogen emissions from air travel are no longer included in the calculations/models, since it is assumed that these emissions will not deposit on the Netherlands, but will instead blow over and deposit somewhere else. This can be hurtful to farmers since emissions from their business are included in the calculations/models and therefore their share of the total emissions might seem overexaggerated. Carsten also informed us that when measuring emissions (from farms), it is important to have proper measuring equipment, from which the lower limit is not too high and therefore unable to measure low concentrations. Moreover, measuring just the concentration of certain gasses, gives an incomplete image since not only the concentration of gasses in barns is important, but also the ventilation rate of the barn.

Next to that, we got more practical and asked Carsten about how ammonia emissions originate on and leave farms. We learned that on a farm level, it is indeed the cattle farms that emit the most ammonia. However, on the level of a barn, the concentration of ammonia might be higher in pigs and poultry barns than in cattle barns. This is because cattle barns in the Netherlands are generally open and have a high ventilation rate since cattle are relatively big, therefore produce a lot of heath and therefore have a big ventilation need. Because there is so much ventilation, any emitted ammonia generally leaves the barn and ammonia concentrations in the barns itself are rather low, around 1-5 ppm. Poultry and pig barns on the other hand are generally closed systems, in which all emissions pass a central point where air scrubbers are used to clean out any leftover ammonia or methane. It is not feasible to use these air scrubbers in cattle barns, since the ammonia concentrations in cattle barns are too low for the air scrubbers to function properly and a high capacity is needed because of the high ventilation rate. Carsten also explained to us that there is an equilibrium between the ammonia concentration in the manure and in the air above the manure. Therefore, mixing the manure with water reduces ammonia emissions from manure, since the mixing lowers the ammonia concentration in the end volume of the manure, therefore less ammonia evaporates to the air above the manure. Moreover, this is the principle behind the closed floors: manure will be trapped under these floors and the air above the manure is hardly mixing, therefore keeping the ammonia concentration in the air rather constant -at high levels- thus preventing extra ammonia evaporations from manure to air. The downside of the closed floors is however that urine will stay on top of the floors (instead of being collected underneath the floor) which causes ammonia to evaporate from the urine on top of the floor.

Next to that, we asked Carsten about the impact of ammonia emissions, and depositions that originate from farms. Carsten informed us that these ammonia emissions are mainly harmful to Natura2000 areas, since those areas are generally sensitive to excess nitrogen. So if ammonia emissions from barns deposit on the land next to the barn, which is generally from the farmer too, this is more than often not a problem. However, if the ammonia emissions travel a bit further or if farms are located close to Natura 2000 areas, there might be a problem. We also asked Carsten if the effect of 1 kg of ammonia is comparable to the effect of 1 kg nitrogen oxide. Carsten informed us that 1 kg of ammonia contains more nitrogen than 1 kilo of nitrogen oxide, and is therefore worse in nitrogen deposition.

Lastly, we asked Carsten what a good solution to the nitrogen crisis would entail and what his opinion is on our project. Carsten informed us that the nitrogen crisis is a complex problem, with a variety of causes/nitrogen emitters. Therefore it is hard, or even impossible, to find a good solution that addresses all the different factors of the nitrogen crisis. Moreover, even if a possible solution would allow the Netherlands to be able to adhere to regulations concerning nitrogen emission ceilings and minimize the harm done to nature, the strict licensing system is rather unlikely to be loosened. Therefore, it would still be hard for farmers to grow and expand their business. Carsten added that this would not be a bad thing when talking about the herd size, but that it should however be possible for farmers to grow in revenue. However, Carsten was very enthusiastic about our project idea and especially liked that we planned to tackle the problem at the very source: by optimizing the digestive system of cattle. Carsten did however question how our feed additive influences the methane production of the cow, since that is also tied to carbohydrate digestion by microbes in the rumen. Moreover, Carsten asked about any possible side effects of the feed additive on the cow, such as rumen acidification. Luckily, we could inform[1]Carsten about the EFSA report we had read previously that stated that similar feed additives had no adverse effects on the health of cows. Lastly, Carsten asked where we would place the MOF. He mentioned that if the MOF is placed in a dairy farm, it has to be able to capture really low (1-5 ppm) ammonia concentrations. And if the MOF would be placed in the top of the barn, to utilize the natural air flow in farms, the MOF should have a very big surface area. We discussed with Carsten what the best place would be to place this MOF and we came to the conclusion that it would be wisest toplace the MOF not in cattle barns, but in the closed ventilation system of poultry and pig barns.

Adjustments:After the interview with Carsten we decided to continue using Emission Registration and Eurostat to find accurate information and numbers about the nitrogen emissions in the Netherlands and the European Union. Moreover, we decided that when we present our project and mention the harm excess ammonia can do to natural areas, we will focus more on the harm ammonia can do to Natura 2000 areas instead of all natural areas, since not all natural areas/plants are sensitive to excess nitrogen. In this way, we are able to give a more balanced overview of the problem at hand. Moreover, even though the sky is the limit for our project, we should keep in mind to be honest and upfront about how much our project is able to, chance, even if everything works out. We will therefore, when presenting our project, include how the current approach to the nitrogen crisis (with a strict regulation system), makes it really hard for farmers to do their work well and that it is unlikely that this regulation system will loosen up any time soon, even if our project is able to help diminish ammonia emissions substantially. And lastly, the most important adjustment to our project after the interview with Carsten is that we will look into placing our MOFs not in a cattle barn, but in the closed ventilation system of pigs and poultry farms, so it can optimally capture ammonia.

Our next steps:To develop our project further, Carsten advised us to look for papers of his colleagues on how well similar feed additives are able to reduce ammonia emissions in cattle. We will search for those and read them. Moreover, Carsten supplied us with two papers that dive more into the ammonia[2],[3] concentrations within cattle barns and how to set up a system to accurately measure this. We will read through those papers as well. Moreover, we will research how many Natura 2000 areas the Netherlands has and how this number compares to the Dutch ammonia emissions and to the rest of the European Union. Lastly, after Carstens rightful remark, we will research how our feed additive influences the methane production of cattle, to see if our project could possibly have any adverse effects on methane emissions.


  1. European Food Safety Authority, “Scientific Opinion on the efficacy of Ronozyme® Rumistar (alpha-amylase) as a feed additive for dairy cows,” EFSA J., vol. 11, no. 10, Oct. 2013, doi: 10.2903/J.EFSA.2013.3434.
  2. J. Vonk, D. van Dinther, J. Mosquera, and N. W. M. Ogink, “Ontwikkel- en validatieprotocol meetinstrumenten voor gasconcentraties in bedrijfsmonitoring van NH3 en CH4 uit veehouderijen : Toepassing bij sensorsystemen voor vaststellen van stalemissies - WUR,” Wageningen Livestock Research (Rapport / Wageningen Livestock Research 1285), 2021. [Online]. Available: [Accessed: 06-Sep-2021].
  3. Commissie Deskundigen Meststoffenwet (CDM), “CDM-advies ‘Stikstofverliezen uit mest in stallen en mestopslagen’ ,” Jun. 2020.

Name stakeholder: Uppsala future farming webinar


Description: The Uppsala iGEM team 2021 hosted a symposium on sustainable agriculture on Sunday the 29th and Monday the 30th of August. While there were many interesting presentations, the one about intellectual property caught our attention in particular. As our project is inspired by a product of DSM, we were wondering how to find out if any patents are violated.

Contribution: The talk went into great detail on how and why to file for patents. Two of the main reasons for filing a patent are to protect your own intellectual property and to put your business in a better position to acquire funding. However, filing for a patent is an expensive process and thus it is important to consider whether it would be cost effective.

Besides being a great way of protecting your own intellectual property, it can also be a great source of inspiration as all patent filings need to be made publicly available. Companies generally do not publish their findings in journals, but instead file a patent application. Therefore these patents contain a lot of technical details on the product they intend on bringing to the market/already brought to the market.

This does leave the question on how to make sure there is no infringement between our product and an already existing patent. Being absolutely sure about this is very difficult. What can be done is searching the database. If a patent which closely resembles our project shows up, we can discuss with the patent office of our university whether the resemblance would actually constitute patent infringement. If no such patent is found, it is however impossible to determine whether we simply missed it or if it doesn’t exist. Therefore, the best course of action would be to just continue with the project as it is. The company that filed a possible patent will let us know if we do end up infringing said patent.

Adjustments: We did not make any changes to our project, but we do have a better image of what we can expect in the future.

Our next steps: We will search for terms describing our project in the database and study any results that show up. If our project ends up substantially resembling an already filed patent, we will contact the patent office of our university to discuss any further steps.


Name stakeholder: Svenja and Irina

Position of stakeholder within company: iGEM alumni and also members of the Safety and Security Committee)


Date of meeting: 16-09-2021

Description: Svenja and Irina were on an iGEM team in 2018. Whilst they were in their own iGEM season and made a project which aimed to degrade electronics waste, they realized that their project could potentially be misused to damage electronics with critically important functions, like a car's navigation system. As their team started wondering about this risk, they came across a term they had never heard before: dual-use research of concern. They were upset that this idea― that some research that intends to provide a clear benefit, could instead easily be misused to cause harm― had never been a part of their education. Hoping to fill this educational gap for other iGEMers, Svenja and Irina designed a workshop to teach other iGEM teams about dual use and how to think about and handle dual use potential of your own project.

Contribution: During the workshop it was first explained that dual-use (research) describes the potential of knowledge or technologies to be used by third parties with both benevolent and malevolent intention. Then it was explained when projects would have more dual use potential than normally and what the difference between biosafety and biosecurity is. All in all, it was very similar to the Biosecurity and Dual-use webinar of the RIVM that we followed in June. However, this workshop did also cover the effect of science communication and how, if it is not done correctly, can lead to public criticisms and panic surrounding science.

During the second part of the workshop, after the explanation, we actually had to work on a case ourselves. During this case we were presented with a synthetic biology project that works with genedrives to prevent the growth of a certain weed. These gene drives could either transfer genes that would make the weed susceptible to pesticides again, or genes that would cause the weed to only produce offspring of one gender, eventually making the population extinct. During the case we ourselves had to identify biosafety and biosecurity risks; how likely it would be for those to occur and how these could be prevented. All in all, this second part of the workshop gave us a concrete chance to practice identifying and mitigating dual use potential of synthetic biology projects.

Adjustments: After this workshop we knew that we were on the right track with working on Safe-by-Design aspects of our project and setting up a framework to prevent as much Dual use potential of our project as possible. So all in all this meetup did not lead to any major adjustments in our project, but it did strengthen our approach that we were currently on: to integrate safety as a key value in one of our projects. And it helped us think deeper about how our own project could have dual use potential: someone with bad intentions might, in the worst case scenario, be able to use our GMO to spread alpha-amylase to the environment which might cause harm. We therefore decided to continue with filling in our WAIR-tool and prepare for our second interview with the RIVM about dual use, where we would present our specific Safe-by-Design choices for our project.

Our next steps:

  • Continue filling in the safety-section of our WAIR tool
  • Meet with the RIVM for the second time to talk about Safe-by-Design for our project
  • Cater our Safe-by-Design approach to specifically prevent/mitigate dual use potential of our project

Name stakeholder: Cécile van der Vlugt-Bergmans and Sam Krouwel

Position of stakeholder within company: Both Cécile and Sam work at the RIVM (Dutch institute for Public Health and the Environment). Cécile is a senior risk assessor of GMO’s and Sam is an Advisor (on the) Education on Safety of Innovative Technologies


Date of meeting: 23-09-2021

Description: As mentioned in the AREA framework written about the safe-by-design interview on the 2nd of August, there would be a second interview to follow up on the lessons learned from the first interview. This AREA framework is dedicated to that second interview. During this interview, we spoke with Cécile, who has been working on SbD and with iGEM teams for multiple years and with Sam, who has a background in responsible research innovation and currently works with SbD and education within the fields of Biotechnology and Nanotechnology. Before the interview, to give an update of what we had done in regards to safety (in all regards of the aspect) so far, we sent Cécile and Sam: some of the safe by design choices[1] we decided to make; the outcomes of the safety section of the WAIR-tool we filled in[2]; our stakeholder analysis[3]; our template for the information and informed consent sheet[4]; and the review of the EFSA on a similar feed additive that we discussed last time and had investigated by now[5].

Lastly, we were given a SbD-case to discuss amongst our team before the interview to learn what taking a SbD approach can entail. The specific case we chose dealt with the risks of a GMO escaping its production facility and the possible added benefit of engineering a kill switch in the GMO to mitigate those risks. We also sent in our feedback on and our conclusions of the case before the interview.

Contribution: During the interview, we first discussed our conclusions of the case. Sam remarked that we discussed and identified a wide range of safety issues, ranging from the possibility of human error to the occurrence of natural disasters such as earthquakes to the possibility of bioterrorism. Sam even mentioned that we came up with ideas that he had never thought of before and asked us if we took the output from the case and implemented it in our own project. We told Sam that we did, and that the risks that we identified, among other bioterrorism and earthquakes (which is a real possibility in Groningen[6]), inspired us to implement a kill switch in our design. That way, if the GMO escapes our large-scale facility, risks would be mitigated.

Cécile informed us that a biocontainment strategy, such as a kill switch, is not always needed as long as standard lab safety procedures are followed. Moreover, in a small scale lab a kill switch is not only not necessary in addition to standard safety rules, but could even hinder standard lab work. We however discussed with Cécile that since we plan on implementing on a large, industrial scale, the risks when a hazard occurs are very different compared to the risk after the same hazard takes place in a small-scale laboratory. Hazards that were discussed in this case were intentional and unintentional release of the GMO caused by human action and natural disasters. We therefore agreed that for our project, which would in the end be executed in a large-scale industrial setting (factory), a form of biocontainment such as the kill switch, would indeed be beneficial.

Both Sam and Cécile urged us to clearly communicate on the wiki which SbD decisions we made and especially during which stage of the project they would be implemented (during the proof of concept, during the industrial production or during the application on the farm). This is because Cécile and Sam both noticed during the interview that we had thought about the safety-issues during industrial production, and how to prevent them, but did not get this from the materials we sent them beforehand.

Sam questioned us about the application and future of our project on the market. He asked that since we designed a novel approach to a problem (fighting gas emissions with a feed additive) that was different from current solutions, if we expected that similar problems would also be tackled with feed additives in the future. We answered that even though we saw a big potential in our project, that we also realised - by using the WAIR-tool - that our project on its own was likely not enough to fight the entire problem and that our project would work the best in combination with other solutions such as urease-inhibitors and air scrubbers. We therefore felt that even though similar problems might in the future indeed be tackled with the help of feed additives, that it was likely that additional solutions would still be needed. Sam then asked us how we saw the cooperation between our project and other solutions. We answered that we would love to make our project work together with other solutions to tackle the nitrogen crisis most effectively, but that in the end the process of the free market would probably play a big role in how this cooperation would be formed. Sam therefore urged us to put this on our wiki as well but at the same time mention that the free market could be flawed and have some risks for the optimal execution of the project as well.

Cécile then questioned us about the review by the EFSA on the safety of a similar feed additive. She mentioned that even though this review mentioned that similar feed additives are safe for cattle, humans and the environment that it would be needed to apply for our own EFSA review in the future. We discussed with Cécile that the current EFSA review investigated the safety of a prokaryotic enzyme produced by a prokaryotic GMO, whereas we are working with an eukaryotic GMO producing either prokaryotic[7] or eukaryotic[8] enzymes. We could still use the current EFSA review as a first indication that our project is most likely safe. However, before actually scaling up our project and ‘taking it to the real world’, it would be wise to apply for our own EFSA review on the safety of our system. Luckily, because of the current EFSA review, we will already have an indication of the kind of data that we would need to gather and supply for such a review.

Lastly, Cécile remarked that she read the take-aways from the safety-part of our WAIR-tool, among which the take-away that we would like to talk to plant experts to discuss any potential adverse effects alpha-amylase could have on starch containing plants. We discussed that even though we might not have the time to do so anymore before the wikifreeze, that it would indeed be smart to reach out to both plant experts and carbohydrate experts to discuss any safety issues related to release of alpha-amylase in nature.

Adjustments: The interview with Cécile and Sam did confirm for us that we are on the right track regarding many safety-aspects of our project. We therefore decided to keep the form of biocontainment (the kill switch) integrated within our project because of the (future) large scale of our project. Moreover, we decided to communicate the safety and SbD choices we made during our project more clearly on our wiki, by allocating an entire section to SbD on our Human Practices page.

Additionally, we decided that before scaling up our project and implementing it ‘in the real world’, we should put our project up for a safety review by the EFSA. And lastly, to develop our project and ensure the safety of our project even further, input from plant scientists and carbohydrate experts would be needed to investigate any risks associated with the (accidental) release of alpha-amylase.

Our next steps: For our next steps we will work hard on the wiki to ensure a clear communication of all our safety and SbD related choices. Moreover, if our project will be continued after iGEM, we will gather the data needed to apply for an EFSA review and apply for this review. Lastly, if our project will be continued after iGEM, we will reach out to plant scientists and carbohydrate experts to identify additional safety risks of our projects and possible measures to mitigate those risks.


  1. For more information: see the part about SafebyDesign on our Human Practices page
  2. For more information: see the part about the WAIR-tool on our Human Practices page
  3. For more information: see the part about our stakeholder-analysis on our Human Practices page
  4. For more information: see the part about our information and informed consent sheet on our Human Practices page
  5. European Food Safety Authority. 2013. “Scientific Opinion on the Efficacy of Ronozyme® Rumistar (Alpha-Amylase) as a Feed Additive for Dairy Cows.” EFSA Journal 11(10).
  6. C. Vlek, “Rise and reduction of induced earthquakes in the Groningen gas field, 1991–2018: statistical trends, social impacts, and policy change,” Environ. Earth Sci., vol. 78, no. 3, Feb. 2019, doi: 10.1007/s12665-019-8051-4
  7. Referring to the alpha-amylase we tested that is originally found in B. licheniformis, B. subtilis or B. amyloliquefaciens
  8. Referring to the alpha-amylase we tested that is originally found in A. oryzae


Position of stakeholder within company: Landscape Ecologist

Date of meeting: 8-10-2021

Description: This stakeholder is a Landscape Ecologist who has contributed to Dutch nature (conservation) policy. This stakeholder has studied plant ecology and landscape ecology and has been working for over 40 years in Dutch nature management and nature conservation. The stakeholder mainly works with improving the connection between vegetation management and water management in nature areas and establishing agreement systems that allow for better consideration of how nature responds to environmental factors. Furthermore, the stakeholder has been involved with Natura 2000 areas and the establishment of the Natura 2000 network. In essence, the stakeholder is involved in writing policy for Dutch nature (conservation) as well as in the practice of Dutch nature (conservation). Therefore the stakeholder was able to tell us both about the practice of nature conservation as well as the policy processes influencing it. Before the meeting, the stakeholder asked us to read through 2 issues[1],[2] containing several articles concerning nature conservation. Based on these articles, we were able to ask more specific questions about nature conservation and the consequences of excess nitrogen/nutrient deposition in nature.

Contribution: During the interview, we learned that nature management has been cut as much as 70% since 2011. These cuts have mainly resulted in less land being purchased for nature management. As a result, fewer nature reserves could be managed to function independently. Indirectly, this has also caused the problem of excess nutrient/nitrogen deposition to increase, since the areas are now unable to function independently and are therefore more sensitive to excess deposition of nutrients/nitrogen. Moreover, nature management itself also suffers from this reduction of monetary funds.

In addition, we found out that many people, especially people living in cities, do not feel a very big connection to nature. Many people don't feel much of a connection with biodiversity and nature conservation in general, but see nature more in the light of the benefits that it provides, for example, recreation or improving health. Much of nature management therefore also focuses on getting people into nature. As a result more people will see the benefit of nature and more people will get involved as supporters for nature conservation. However, children, unlike adults, are not necessarily interested in "utility nature" but are often intrinsically amazed by nature.

Furthermore, we asked the stakeholder how big the problem of the nitrogen crisis really is. Our stakeholder informed us that in ecology and nature conservation it is all about coherence: excessive nitrogen deposition is not the only problem for nature. The stakeholder therefore prefers to talk about an excess of nutrients, rather than an excess of nitrogen alone. Ultimately, what matters is the availability of nitrogen, phosphate, or potassium in the soil, sludge, or water. The form in which the ion occurs matters, of course, but so does the ratio of these substances. Moreover, most of the time emissions to the air are mentioned, but air (like water) is primarily the means of transport for these substances. Nitrogen is currently in the spotlight because action groups in the Netherlands have gone to court and demanded legal consequences to ensure that the state does address the excess of nitrogen emissions. Other nature conservation problems would also be addressed if action groups went to court and enforced legal actions. The stakeholder, however, informed us that this is not desirable because then each factor/problem would be treated individually, while in practice the coherence of factors is much more important for an ecosystem. However, the stakeholder does note that since nitrogen emissions have been reduced since 1980, both the stakeholder and colleagues have noticed positive effects: certain species that are sensitive to excess nitrogen and whose abundance had declined, are now returning. There is still much to be done, but reducing nitrogen emissions certainly has positive effects on nature.

We also talked to the stakeholder about the importance of Natura 2000 areas in the discussion of excess nitrogen/nutrient deposition. The stakeholder informed us that besides the Natura 2000 areas, there are also other Dutch nature areas that are sensitive to excess nitrogen/nutrient deposition. However, Natura 2000 areas are legally protected at the European level, and the other nature areas are not. So legally there may not be a problem for these areas, but in practice there is. In fact, often these areas are already not doing very well, even without excessive deposition of nitrogen/nutrients, which is often why these areas are not designated as Natura 2000 areas.

We then discussed alternative solutions to the nitrogen crisis with the stakeholder. One of these measures is to relocate farms to ensure that they were further away from nature reserves and thus, hopefully, less nitrogen/nutrients would be deposited in these nature reserves. However, the stakeholder raised some questions about the models that support this. At the time these models were created, finances for the measurements that would support these models had been severely cut back. As a result, there are still many uncertainties in the models and it is thus questionable whether moving farms would have the desired effect. However, it is certain that the Netherlands, certainly compared to other European countries, emits far too much nitrogen and that something needs to be done to combat this. The stakeholder thinks that on a policy level, it is unavoidable that the number of livestock must be reduced, among other things to reduce nitrogen emissions but also because the current bio-industry with low meat prices has a large, uncompensated, burden on the environment. Secondly, we asked the stakeholder about the feasibility of sod-cutting natural areas to counteract the negative effects of excessive nitrogen/nutrient deposition. The stakeholder informed us that this measure only has added value, sometimes, in heather fields, which are a rather uncommon nature area. However, sod-cutting can also release other nutrients stored in the soil, which can also have strong adverse effects on an ecosystem. Moreover, it is questionable whether it is fair that only in the nature reserve an excess of nutrients should be solved that was created/released somewhere else.

Finally, we talked to the stakeholder about our project and any potential problems it might cause. The stakeholder asked us about the safety of our feed additive for cows. Despite the fact that -understandably- it is not possible within iGEM to test our feed additive on cows, and similar feed additives3 have been shown to be safe for cows, the stakeholder recommended that if our project were to continue beyond iGEM, we should investigate the effect of the feed additive on different breeds of cows and the effect on the composition of the microbiome of cows. The stakeholder brought this up since the feed additive interacts with the microbiome of cows, which is different per cow breed. Moreover, it might be possible that the composition of the cow's microbiome could change as a result of this interaction. In addition, the stakeholder emphasizes that, despite the kill switch and the fact that the GMO would not leave the plant/lab in our design, it is very difficult to oversee all the safety risks of working with GMOs. Therefore caution is warranted to prevent the GMO from entering the outside world. Furthermore, the stakeholder informed us that it is good that we plan to combine our project with other techniques so that we do not tout our project as the only and complete solution to the problem. The stakeholder also stressed that our project should be financially and practically feasible for farmers at the end: too often in the past, (small) farming companies had to invest large sums of money for techniques that in the end were no use at all or were even flawed, causing fires in farms.


Based on the interview with the stakeholder, we decided to adjust our project in the following manner:

  • We will communicate clearly that even though reducing nitrogen emissions is beneficial for nature and biodiversity, it is not the only problem that endangers nature. In the end, nature conservation is all about coherence.
  • Even though we have already understood from previous stakeholder interviews that Natura 2000 areas are important within the discussion of the nitrogen crisis, we will no longer focus our project completely on protecting only Natura 2000 areas. We will still mention Natura 2000 areas as being important in the discussion, but we will mainly focus on the harmful effects of excess nitrogen/nutrient deposition for all the nitrogen-sensitive nature areas.
  • Up to how far it is still possible within the time we still have left for the iGEM competition, we will advocate for the (financial) investment in the legal protection of sensitive nature areas, also those that do not (yet) belong to the Natura 2000 network. Moreover, we will advocate for a more coherent approach to environmental protection in which the focus is on multiple factors affecting nature, rather than just one factor. In addition, we will advocate for getting people involved (through volunteering or recreation) within nature to strengthen the support for nature conservation.
  • If our project is pursued beyond iGEM, we will commit to/recommend research on the effect of our specific feed additive on the microbiome of different cow breeds. In addition, we will commit to/recommend doing research on the exact (financial) benefit and possible risks of our project for farmers, such that our project will only be marketed if it is actually (financially) beneficial for farmers and does not cause safety risks.

Our next steps:

  • Communicate clearly on our implementation page on the wiki, that our project is not the complete and only solution to the nitrogen crisis but that a coherent approach is needed in which multiple solutions are combined and more problems are tackled than just the excess of nitrogen emissions.
  • Communicate clearly everywhere on our wiki that the excess of nitrogen emissions is a problem for all nitrogen-sensitive areas, some of which are Natura 2000 areas, and that reducing those emissions has positive effects on nature.
  • Advocate: for legal protection of sensitive nature areas; a coherent nature protection approach; getting people involved in nature; not using our project in real life before its effect on the microbiome of different cow breeds is known; not using our project in real life before the exact (financial) benefit and the exact risks for farmers are fully known.


  1. De Levende Natuur Issue 2 of 2013 | Theme: Toekomst Voor de Natuur.” 2013. De Levende Natuur. (October 8, 2021).
  2. De Levende Natuur Issue 3 of 2013 | Theme: Toekomst Voor de Natuur (Part 2).” 2013. De Levende Natuur. (October 8, 2021).
  3. European Food Safety Authority. 2013. “Scientific Opinion on the Efficacy of Ronozyme® Rumistar (Alpha-Amylase) as a Feed Additive for Dairy Cows.” EFSA Journal 11(10).