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 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.

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

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.

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

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

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Date of meeting: 21-6-2021

Description:

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.

Contribution:

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.

Adjustments:

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

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Date of meeting: 23-6-2021

Description: 

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.

Contribution:

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 (rcc@igem.org), Human Practices (humanpractices@igem.org), Safety & Security (safety@igem.org) or Diversity & Inclusion (diversity@igem.org). The potential awards given by these committees were also brought to our attention.

Adjustments:

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 

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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 

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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

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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.