‘Never ignore a gut feeling, but never believe that it’s enough.’ - Robert Heller
Introduction
We have all come across it at one time or another - that gut feeling about something that is just right. It might have been about a person you really like, an investment, or a biosynthetic solution that might revolutionize viral disease prevention by solving a worldwide crisis. Still, it is very important not to get tunnel visioned by your emotions. Always critically assess whether this so called righteousness is actually grounded. This person might have some major red flags, the company might be on the verge of bankruptcy; -or your solution might have major negative impacts on the environment or the health of its users.
On our human practices page, project description page, and home page we have shown that the avian influenza crisis is a very complex problem involving various stakeholders with different, and sometimes even conflicting interests and needs. It is a complex problem, whose solution requires the careful consideration of the perspectives of involved stakeholders and the safeguarding of values such as safety and responsibility. Only by understanding the problem to the fullest extent can you truly create a safe, responsible, yet effective solution that will make the world a better place for everything living in it.
It is crucial that we use the data we gather in our human practices approach to improve our project in a structured and organized manner. Only then will we be able to properly implement our new found insights into our solution. To accomplish this, we used the AREA Framework for Responsible Innovation, as introduced by iGEM Exeter 2019 (figure 1), as explained on our human practices page. Using this framework, we wrote out summaries of all of our interviews and human practices activities, thereby structuring our integrated human practices in a very organized manner in the process. The first part of the AREA consists of the ‘description’, where we described the reason why we conducted the interview, the expertise of the interviewee, and other information that we thought was important to mention. The second part of an AREA summary consists of the ‘contribution’: "What did we learn from the interviewee? This can range from feedback on the approach of your solution, to insights on the problem of avian influenza itself. In this part we described what the stakeholder added to your pool of knowledge. The third, and perhaps most important part of the AREA, consists of ‘adjustments’: how did we change our approach/solution as a result of the interview. This can range from changing the implementation of your solution, to changing experimental/fundamental designs for your solution as a result of the interview with the stakeholder. In the last piece of the AREA summary, we laid out our ‘next steps’, which included the steps that we were going to take to make the previously described adjustments a reality.
On our human practices page we extensively described the methods we used to interact with stakeholders, gather feedback, learn more about the problem, and what we ultimately did to find out how we can make our solution responsible and implementable in the ‘real-world’. On this page, we will describe what we learned from our human practices approach, by using the AREA framework (figure 1), and how we ultimately used this data to create a solution which has a positive impact on the world.
Figure 1. The 4 steps of the Anticipate, Reflect, Engage, Act (AREA) framework including the extra layer iGEM Exeter 2019 introduced: description, contribution, adjustments, and our next steps).
We used the AREA framework to integrate all the aspects of our human practices approach into improving our project. This therefore also includes the literature review, survey, stakeholder analysis, and collaborations/meet-ups with other teams. During these meetings, we also gained valuable feedback which would later shape our project and our planned experiments. We decided to treat the survey as a sort of ‘interview’, which provided the perspective of the general public. These AREA frameworks were then used to improve our project; figure out next steps in our potential implementation; and define our key values, which can be found back on our project description page. Our integrated human practices also helped us develop our safe-by-design approach which can be seen on our safety page. Our integrated human practices not only perfected our final solution and its key-values, but also assisted in improving the human practices work itself, thereby integrating our integrated human practices work back into our original human practices approach. This way, we learned how and who we could contact through well-connected stakeholders. Stakeholders also thought us where and how to gather useful information regarding the problem in literature. The stakeholders also provided us with knowledge which is not widely available, which we were then able to integrate in our question list for use in future stakeholder interviews . A good example is our stakeholder interaction map, found on our human practices page, which was regularly updated after stakeholder interviews. We also then continued to present our updated map during the next interview for new insights.
To sum it up, the literature review, stakeholder & expert interviews, collaborations, and stakeholder analysis formed the pillars and foundation of our human practices work as seen in figure 2. Using the AREA framework, we built upon this foundation to create our integrated human practices. Thanks to this specific approach, our integrated human practices both adds towards the solution and project, as well as to the human practices approach it originated from. Through this circular human practices approach we have not only been able to optimize our stakeholder analysis, but we have also been able to organize more interviews than we would have otherwise through our ‘stakeholders-know-more-stakeholders’ strategy. Here we always asked whether stakeholders could provide us with people that we had not previously thought of, or simply asked them to send our details to colleagues in other fields that might be interested. We have schematically depicted this approach in figure 2. Our integrated human practices work shows how we were able to identify our key values, and how we integrated them into our final project: Nanobuddy.
Figure 2. The pillars which form the foundation of our human practices, which are used to define our key values which would later shape our project through our integrated human practices. Created using Biorender.com
Insights from our literature review
Contribution
We conducted a literature review on the avian influenza crisis. To be specific, we focussed mainly on the avian influenza outbreaks in the poultry industry. We found that the number of outbreaks of avian influenza is rapidly increasing, not just in The Netherlands, but across the whole world. It appears that vaccination strategies are currently not delivering the results that are needed to properly protect poultry on a large scale. In addition, vaccination strategies are prohibited to implement because of EU regulations. At this moment, it appears that there is not going to be any big policy change. Therefore, farmers and local governments will have to rely on biosecurity measures to keep their poultry safe. Besides all this, the poultry in The Netherlands by far forms the largest proportion of livestock by numbers. This also means that we would have to regularly vaccinate one hundred million poultry in The Netherlands alone. While people are pleading for a solution other than biosecurity, which has proven to be insufficient as of yet, there simply is not one available. With vaccinations unable to properly protect poultry, and regulation prohibiting implementation of them completely, the only thing farmers can/have to do is to cull their flocks when there is an infection. This problem seems more intricate than we at first thought. The poultry industry is very complicated and tens of years of policy are involved prohibting a lot of the proposed solutions we have now. It will be very difficult to create a suitable solution. Check for a more detailed problem description on our project description page, which also includes our references!
Adjustments
Although we at first thought about tackling the problem at the (potential) source, we quickly figured that it will be very hard to find a biosynthetic solution that can be implemented in wild animals. Therefore, we decided to put our main focus on the poultry industry, which arguably is also the place that suffers from the most damage as a result of this epidemic. We have thought about an immunization strategy where we would engineer a GMO as feed additive, with the ability to secrete virus particles which could consequently immunize the chickens. We could also spray these bacteria into the air, upon which they would habituate the lungs and gut microbiome of chickens. However, literature suggests that this might not be as effective in the real world, since you would not produce enough virus particles to initiate a proper immune response for the highly pathogenic variant. Therefore we decided against this strategy. Still, we appreciated the spray-aspect of our previous strategy, so we kept on brainstorming on how we could utilize microbes to create a spray-based vaccine. This is especially relevant since it allows for scalability, which is a known bottle-neck for conventional intravenous vaccinations. Additionally, this problem is relevant and local to our own community. Therefore we decided to dedicate our iGEM project to addressing this terrible epidemic. It was at this point that we read up on nanobodies which could be sprayed into animal lungs to prevent them from getting infections from viral disease in vivo [1]. We then came up with our main plan, and decided to further develop it into our iGEM project. This plan consisted of having a microbe that could secrete nanobodies, and placing it in the chicken lungs through a spraying system. While a lot of the details still need to be filled in, we are confident that this plan is something we could work with in the future.
Next Steps
Figure out what microbe is ideal to use for a host organism: one that can be genetically modified to produce nanobodies. Ideally it also lives in the chicken lung microbiome naturally making our proposed plan more safe for the chickens.
Find nanobodies that can bind to avian influenza.
Brainstorm on experiments that we could conduct to test our theories.
Create a human practices framework based on our new found ‘main topic’: stakeholder analysis.
Conduct more background research.
Focus the story of our project on the avian influenza crisis in The Netherlands; specifically on the poultry industry.
Insights from our stakeholder analysis
Contribution
On our human practices page we have described our stakeholder analysis. This stakeholder analysis gave us the opportunity to have a good brainstorming session on who is actually involved in our problem. We quickly realized that most of the stakeholders that had the highest involvement/interest, also appeared to be the stakeholders that were the most affected. These stakeholders located in the top-right and bottom right corners (stakeholders that should be useful to our project) are often involved in the poultry industry, NGOs, and the government. They are either responsible for dealing with, or are affected by, the avian influenza crisis. Besides these institutional and individual stakeholders, there is also the poultry themselves. Sadly the poultry themselves have been depicted among the highest interested groups, with the lowest power among all of them.
We also noted that governmental bodies dealing with public health and waterworks are not that invested in avian influenza, as it is not spreading to humans yet. Still, the governmental bodies that are responsible for dealing with avian influenza in the food industry are very involved in the problem. These governmental bodies all share a lot of power, with very divided interests in the problem. The pharmaceutical companies, with arguably the largest power among all of them, are depicted in the middle of the interest spectrum. This is because they are waiting for a change in policy before they will act, while still trying to address the problem where they can. This stakeholder would definitely be interesting to interview in our human pratices.
Adjustments
The pharmaceutical industry is an interesting stakeholder because they have a lot of experience with providing solutions to these types of problems. Now, the problem is that their hands are tied which does not stimulate them from investing in proper solutions to the avian influenza epidemic. The first thing we want to do when we get our human practices approach up and running, is to speak to this stakeholder with high priority. We used the stakeholder analysis to give us a framework to use when weighing the different needs and wishes of stakeholders when designing our eventual solution. We will consult this stakeholder matrix every time we have conflicting interests between stakeholders with high power, or high involvement. We also have plans to create a stakeholder interaction map, to further show the interrelationships between stakeholders. We believe this could add to our original stakeholder analysis from our human practices approach. On top of that, we think it is important to keep a close eye on the networks shared between stakeholders. Stakeholders often know of each other, and work together with similar people. We use this to our advantage, and conduct more interviews using their network containing of involved stakeholders.
Next Steps
Reach out to pharmaceutical companies as soon as possible. With a lot of power, these companies could be very influential in a project such as ours. We need to find out whether our solution is something they would consider themselves, and whether they can provide us with valuable feedback
Figure out a way we can use the network of stakeholders to our own benefit.
Write down how we will manage to actually ‘actively engage’ with the stakeholders in the top-right corner of the stakeholder matrix (besides doing interviews).
Start gathering contact information of stakeholders. As soon as our safety and security precautions are ready, we can start contacting them.
It is important to consider the chickens as an important stakeholder. Even though they do not have a lot of ‘power’ in our analysis, they might be our most important stakeholder yet. Just because individuals, industry, or institutions have more power and about as much interest, we should not create this solution just for them. This would simply be unethical. Therefore, we decided that one of our key values should be ‘animal health’.
Change our project goals in accordance with our newfound key value; preventing the unnecessary culling of poultry, preventing poultry from getting disease, potentially preventing a next pandemic, and saving the poultry industry money in the process.
Insights from our stakeholder interviews and collaborations/meet-ups
As explained above, and on our human practices page, we have consulted numerous stakeholders to gain valuable feedback on our project and deepen our understanding of the avian influenza crisis. In addition, we attended meet-ups and participated in collaborations where we learned about aspects such as safe-by-design, or gained valuable feedback from peers. Together, all of this feedback had a large influence on the final design of our project. We reached out to all relevant stakeholders as defined on our human practices page stakeholder analysis. To give a brief overview, we reached out to the following main groups of stakeholders depicted in a stakeholder landscape in figure 3.
Figure 3. Brief overview of the groups of stakeholders that we consulted in a stakeholder landscape.
Because of the framework laid out by previous iGEM teams we were able to use a quality AND quantity approach with regards to our stakeholder approach. In the timeline below, we have included all the summaries outlining how our stakeholder interviews and other human practices activities contributed to significant change in our project. Besides that, it outlines who we spoke to and what we learned from each individual stakeholder. As explained on our human practices page, we consulted stakeholders about the problem and impact of avian influenza, how they were involved in the problem, and for feedback on our solution. For a detailed overview of what we asked our stakeholders specifically, check the general interview outline on our human practices page..
June
Date of meeting:18-06-2022
Description
We attended the first workshop organized by the iGEM working group providing a series of workshops on different aspects of the competition. There were also some ambassadors present who shared their stories and how they ended up becoming an iGEM ambassador. The head of the human practices program gave a
presentation on how to properly conduct human practices as an iGEM team. We used this workshop to try
and figure out how we would further shape our human practices approach. We already put in some efforts
towards responsibility and safety, but we try to figure out where we can still improve upon. We were
especially interested in the data-security and informed consent which is needed before stakeholder
interviews. Also, this workshop was a nice opportunity to gain some inspiration from the human practices
team.
The theme of the presentation was: ‘how to do human practices in your project-life cycle’.
Contribution
We learned about key concepts in the general human practices framework: be reflective, be responsible,
and be responsive. We have not yet heard of these so called ‘3 Rs’so we were eager to learn more. These
key concepts touched upon the things that every human practice work should focus on. This included that
you should think of values and needs you are prioritizing, and where you are compromising. Also, how
you can communicate honestly and consider how your project could impact the world, for better or worse.
Lastly, you should listen to your stakeholders, and learn as you engage with them. The human practices
team showed us a lot of good examples of different human practices from successful teams. We were
happy to see that these teams often did things that we were planning to do as well. We especially liked the
advice the presenters gave on how to find and contact stakeholders. The last advice that we tried to take to
heart was to not just look at The Netherlands, but to go beyond our own country. For us this is very
important since the problem we are addressing is not just a Dutch problem, but a worldwide epidemic.
Adjustments
To address the 3 Rs, we chose to have a brainstorming session on how we can address these key concepts
of iGEM human practices work. As a result of thinking about how we can address responsibility, we
decided to think of key values that we could attribute to our project. This could steer our project to
become a better version of itself. We also decided that we would change these key values if that would be
necessary. At this stage it is difficult to say what our project is gonna look like exactly, therefore our key
values are prone to change. Especially since we are planning on talking to a lot of stakeholders, which
also nicely addresses one of the 3Rs: Responsiveness. The workshop confirmed to us that we want to put
most of our focus on stakeholder engagement. Especially since the problem we want to address is such a
relevant problem, we are wondering what they will think of our novel approach to solving this issue. We
also decided to look into facilities within our university to investigate whether there are ethical
committees and data security officers which might be able to help us with our safety and security.
Our Next Steps
Do research on how previous teams created their informed consent sheets.
Review the safety and security work we have done so far with the new knowledge learned in the
workshop in mind.
Think about how we can be: reflective, responsive, and responsible throughout the process of
conducting our own human practices.
When working on future human practices activities, keep in mind the defined key values.
Date of meeting: 24-06-2022
Description
We got the opportunity to have two meetings with the RIVM to discuss safe-by-design aspects in our
project. The RIVM is the National Institute for Public Health and the Environment, a governmental
organization in The Netherlands. This meeting was an excellent opportunity to discuss our preliminary
design and further steps we could take with both a relevant stakeholder, as well as experts in the topic of
‘safe-by-design’. After our presentation on our plans, the RIVM gave us valuable feedback and taught us
things about safe-by-design principles.
Contribution
After our presentation there immediately were a ton of questions regarding several aspects of our project.
One of which was related to future perspectives: how would we test this and come to a proof of concept?
As we had a pretty good idea of our experimental setups, we discussed how we could take this project
further and implement it in the ‘real-world’. These steps would include the so-called DM1, DM2, and
DM3 steps. Here you would do tests on a small scale population of chickens, and build up to test it in a
real farm. The RIVM provided us with contact information of a big pharmaceutical company that has a lot
of experience in this type of animal research.
The primary thing we discussed was safety. Mainly, we talked about our biocontainment strategy. We had
read some literature on kill-switches that make use of CRISPR-Cas, which we talked about. We did not
yet specifically know what the triggers would be of the kill-switch though. We talked about how it could
work, and what inputs (kill signals) we can use. An interesting thing that was named is light, which we
had not thought of before. Temperature was a relatively obvious one which the RIVM agreed on. We also
briefly addressed deathman kill witches, and making our bacteria dependent on specific substances such
as non-canonical amino acids which can not be found in nature. These might be interesting to implement
in our project as well.
The RIVM also introduced us to the safe-by-design framework, which we had previously not considered
in our project. We talked about how we can incorporate safety in our project. A good example of this, is
the use of a Generally Recognized As Safe (GRAS) bacteria as a host organism for our solution. Luckily,
the bacteria that we wanted to use has the GRAS label, which is good news! The RIVM also spoke about
how other steps in our solution can be adapted in order to make them more safe.
Adjustments
The RIVM told us that it is very interesting to still include certain research that we did not do, but did
think of. A lot of iGEM teams do not explicitly do this, which they think is a pity as it shows how a
project might still have a lot of potential, even though you did not manage to conduct all of your research.
We therefore decided to take this advice to heart, and try to include as much of our research (including
potential hypothetical steps) on our final webpage.
We have already conducted some in depth research on non-canonical amino acids and deathman kill
switches. Both have their advantages and disadvantages to the temperature system we have proposed.
However, since we would want our strategy to be as convenient as possible (one spray and poultry is
protected), we decided against non-canonical amino acid approaches since you would need to
continuously spray these substances into poultry lungs which is not practical. Deathman kill switches are
not suitable for our applications because our microorganisms should be killed immediately when escaping
the lung microbiome.
As a result of the talk we had about kill-switch, we have decided to create a 2 input OR-gated killswitch
which uses light and temperature. For the temperature we already have some preliminary designs, and we
will try to incorporate a light switch within the same pathways. This can improve our biocontainment
strategy tremendously, as escape frequencies often are the biggest problem with kill-switches. The RIVM
really pressed on the importance of a good working kill-switch. Therefore, we will try to design the best
one that we can think of in order to make our project as safe as possible.
Our Next Steps
It would be really interesting to speak to the pharmaceutical company that the RIVM named for
more feedback, and ways we could actually implement our research in the real world. We have
already initiated contact and are now waiting for a response!
Research and design our new, extra safe, kill-switch with 2 signal inputs. Although we have no
knowledge of researchers actually getting a 2 OR gated killswitch to work, we will still try as it
could bring the safety aspect of our project (one of our key-values) to a whole new level.
Implement the safe-by-design framework. This also connects to our key-value ‘safety’ which is
one of the most important out of the bunch. We will create a safe-by-design plan which explicitly
states how our project is safely designed from beginning to end; for everyone involved.
July
Date of meeting: 01-07-2022 - 03-07-2022
Description
The iGEM Hamburg team organized together with the German Association for Synthetic Biology
(GASB) an European meet up. The goal of the meet up was to find out about the projects of other teams
and to see if common ground could be found for collaborations or partnerships. Over the weekend there
were multiple speakers and workshops organized with the goal of self-exploration or the larger framework
wherein iGEM functions (e.g. workshops about biosafety). Each team also gave a short pitch to introduce
their project and this was accompanied with a poster session.
Contribution
During the meeting we got into a conversation with the iGEM team from Wageningen. During this
conversation we discovered similar grounds for our plans to develop a kill-switch based on a temperature
trigger. At the European meet up we began discussing the possibilities of a collaboration.
Adjusments
Initially we planned on doing a collaboration with another iGEM team from The Netherlands based on
common grounds of the Human Practices part. But due to the coincidentally similar approach of the
Wageningen team we decided to focus on collaborating with the kill-switch project.
Our Next Steps
Currently we are working together with Wageningen with the approach that we validate their
constructs and they ours. Additionally where we can we’d like to exchange data to see if similar
results are obtained from different labs. This collaboration seems very promising so far and it
might lead into a partnership later on!
Date of meeting: 08-07-2022
Description
The Centre for Living Technologies organized the Dutch iGEM meet with the support of iGEM WUR and
iGEM TU/e.This event provided a common platform for the Dutch iGEM teams to come together,
network, and explore collaboration. Keynote speakers Randy Rettberg (President of iGEM foundation)
and Kyra Delsing (Rathenau Instituut, The Netherlands) spoke about what iGEM has to offer. All dutch
teams also presented their project and research so far, this allowed for nice moments for feedback from
fellow researchers and their supervisors. The day ended with a brainstorming speedround session, in
which participants shared experiences and advice.
Contribution
During the meeting our partnership with Wageningen solidified, we talked about the future steps of our
research together. Furthermore, our team exchanged many sponsorship experiences with all teams present.
We were surprised to see a stark difference in approach to raising financial means to do research.
Furthermore, we asked around if anyone knew a stakeholder involved in farming, since we at that time
were trying to fill this gap in our stakeholder map. Additionally, we provided other teams with contact
details to municipalities, governmental bodies and other politically active stakeholders.
Adjustments
Even though we have found research on the lung microbiome of chicken which shows L. reuteri present,
we got a question about the research on the lung microbiome of chickens local to The Netherlands. More
specifically, chickens within the poultry industry, as living conditions (and therefore microbiomes) are
very different between chickens in a research setting and chickens in the poultry industry. It is very
important that this microbe is native to chicken lungs in the poultry industry, as this will indicate that our
solution will most probably be both safe and effective. To increase our knowledge on the safety of our
project we decided to look into this. Confirming the presence of L. Reuteri in dutch chicken will give us
more certainty of our Nanobuddy not being harmful to the chicken it would live in.
Additionally, we got in contact with a German poultry farmer through a member of another dutch team.
Our Next Steps
We are changing our approach to looking for sponsors from a sponsor-packages approach to an
open approach aimed towards a conversation rather than a standardized sponsor agreement.
Finally, another important next step is conducting a microbiome analysis of Dutch poultry lungs.
Date of meeting: 11-07-2022
Description
Stakeholder is involved in tackling the avian flu pandemic through a position as policy advisor with a
background as veterinarian. They work together with Dutch government in order to do this. We discussed
all aspects of the avian flu pandemic in The Netherlands from the stakeholders perspective. This ranges
from the impact this pandemic has on the relevant stakeholders, to the policies and systems in place to
tackle this problem. This, gave us a broader perspective on the problem, and on potential other
stakeholders to talk to.
Contribution
After the interview with the stakeholder, we have broadened our knowledge about the problems causing
and arising from avian flu in The Netherlands. An avian flu outbreak can have two origins: through wild
birds infecting poultry, or between-enterprise contamination. We talked about the systems which are in
place now to prevent outbreaks in the poultry industry: biosecurity and culling. Although there is a lot of
energy and money invested in biosecurity, culling of poultry happens more and more frequently.
Stakeholder expresses that this has both a heavy financial and emotional toll on everyone involved. While
there are numerous measures for tackling avian influenza in the poultry industry, little to nothing is in
place to prevent infection of wild bird populations. Stakeholder states that it seems to be that biosecurity
is not enough to keep the virus out of the poultry industry, even though the biosecurity of the Dutch
poultry industry is of a high level. We also talked about the risk that the virus might mutate to become
specific to humans, which remains something to be wary about as it might just go wrong if we do not act.
Stakeholder expresses that we cannot in good conscience keep on going with the current status-quo. They
express that all of the sectors agree that we have to find other ways to prevent disease spread. According
to the stakeholder the biggest obstacle in protecting poultry through vaccination are trade regulations in
the EU. The Netherlands is a big exporter of poultry, and vaccination will result in trade hindrance.
Adjustments
The stakeholder explicitly says that it will be crucial for our product to not leave any GMO remains in the
meat. This will be both important for export, but also EU regulations with regards to safety. They say that
it is important that if we want to create something that will actually be used, we have to actually improve
upon conventional vaccination. Therefore we will have to focus on what conventional vaccinations lack:
affinity for mutated strains and up scalability. Also, the costs appear to be very important: it is important
that it can be cheaper than conventional vaccination.
Our Next Steps
Contact relevant stakeholders involved with wild bird populations and avian influenza.
Think of people we could contact which are involved in EU regulation.
Think of whether we can conduct an ethical analysis on how our solution might compare to
conventional vaccination.
Check current regulation with regards to GMO usage in the animal industry.
Talk to stakeholders to find out the costs of vaccinating, and not vaccinating poultry.
Date of meeting: 13-07-2022
Description
Stakeholder is involved in infection monitoring, financial aspects, and policy regarding avian influenza.
Stakeholder is also involved in talks with EU regarding avian influenza. We used this interview to gain
the perspectives of the poultry farmer sector on avian influenza. We were also interested in the impact the
pandemic has on poultry farmers themselves. This includes both emotional, and financial damage.
Contribution
In the monitoring system’s poultry farms are periodically checked for avian influenza through random
sampling. Through a early warning system poultry keepers also keep an eye on symptoms. If there
appears to be an infection, the veterinarian is called in for an inspection, and samples are sent in for an
analysis. If there is an avian influenza outbreak, the NVWA is called in and will deal with the
measurements which are in place to prevent further spreading (e.g. culling, poultry travel restrictions,
caging up poultry in the area). Stakeholder expresses that the biggest impact is for the poultry farms; both
tangible and intangible. Culling of the animals takes a heavy toll on farmers themselves, as most of their
birds at an early stage of an outbreak are not even infected. These farmers will also have to see their
healthy poultry die. The culling of a farm, depending on size, costs a farmer around €500.000 to
€1.000.000. This is paid by the entire sector up until €35.000.000 in total, and is not insured. So the sector
pays for this as a collective. If it goes over this amount, the government pays the rest.
There are still a few steps in biosecurity that can be taken to more effectively prevent disease spread.
However, the stakeholder expresses that the systems in place are already functioning highly efficient,
whilst the frequency of avian influenza outbreaks still increases. There are two main problems: tackling
the source, and vaccination policy. We cannot prevent birds crossing to The Netherlands carrying the
disease and spreading it among poultry and themselves. This is also the place where the disease mutates,
spreading to other wildlife such as foxes and possibly humans in the future. Vaccination becomes more
generally accepted as the pandemic evolves. Through the years this epidemic has impacted numerous
people involved in the industry in various countries. Therefore, countries are more accepting of
vaccinations in general. However, it will still take a while to make space for them in terms of policy.
Adjustments
The DIVA-principle is important for vaccines in the animal industry, as well as still being able to
primarily prevent against spread of the virus. To comply with trade regulations you must be able to
discriminate between vaccinated meat and infected meat. We will have to think on how that might impact
implementability of our solution. The stakeholder underscores that prevention in wild bird populations is
also crucial, besides vaccination of poultry. We should investigate what the possibilities would be through
talking to relevant stakeholders to potentially implement in our project, as tackling the source (wild bird
population) eventually is most important. If we were to actually implement this, we should really focus on
biosafety. Poultry farmers are still food producers, and therefore it is crucial that everything is safe. We
will have to give extra care to GMO containment, well-being of chickens, and food-safety. We should
also realize that with any vaccination strategy (possibly including our alternative solution), there is still
intensive screening on whether the virus is not silently spreading among the protected chicken population.
Considering the nature of our plan and approach, this might not have to be the case for our strategy. We
will have to further analyze and verify this before we can properly adjust our plan of approach.
The stakeholder told us that it would be interesting to individual poultry farmers and other stakeholders
which we were not aware of, and helped us on the trail to get into contact with them.
Our Next Steps
Talk to individual farmers in order to gain their perspectives on our solution.
Investigate the impact of our solution on food-safety and chicken well-being. Talk to experts and
look into general food law.
Think of ways how we can improve our biocontainment strategy.
Date of meeting: 18-07-2022
Description
Stakeholder works in various committees on tackling and monitoring avian influenza in wild bird
populations, with a particular focus on the North Sea Region. In this interview we were particularly
interested in the perspective of people involved in avian influenza outside of the poultry industry.
Contribution
Avian influenza has been around in bird populations for a while now, however, the highly pathogenic
variants (HPAI) we see now are a result of the massive poultry industry. This has also a big impact on
people in the field of avian wildlife (e.g. population monitoring and counting).
Where the poultry industry tries to keep avian influenza outside of the industry, bird wildlife conservation
organizations try to obtain the opposite. The highly pathogenic variants that originate from the poultry
industry can infect wild bird populations with detrimental consequences for wildlife. Stakeholder
mentions that vaccination in the poultry industry will be of great importance in order to prevent this.
Although it has already jumped to wild bird populations, it is clearly important that we act quickly to
prevent further spread and mutation of the virus worldwide.
It will be difficult to actually do something for wild birds (e.g. vaccination). However, there are talks with
the relevant ministries about nation wide measures to prevent spread among wild birds. Stakeholder is
also involved in talks to tackle the problem at the source: the intensive poultry industry.
The stakeholder also pointed us to the ‘Nationaal Actieplan Versterken Zoönosenbeleid’ (National Action
Plan to Strengthen Zoonoses Policy) which touches upon the strategy to prevent avian influenza from
mutating to humans. We also asked how you might prevent wild bird populations from getting infected.
The stakeholder underscored that targeting the source, the poultry industry, will be most effective.
Measures like removing affected wild birds from populations can also be effective to slow the further
spread to other birds, but in most cases it would already be too late, and this by no means is a solution to
the prevention of new outbreaks of HPAI. Vaccination of wild bird populations will be very unlikely to
impossible according to the stakeholder.
Adjustments
After our conversation we quickly realized that measures to prevent spread and infection among wild
birds can be strictly through a change in the densities at which poultry are kept, since this leads to the
evolution of HPAI. There is no possibility in the near future to tackle this pandemic by targeting wild
birds themselves. We will have to keep our focus on the poultry sector specifically with our solution.
Biosecurity in the poultry industry can be improved, but will not be enough. The stakeholder also
expresses that there are also sustainable ways of preventing uncontrolled spread of the virus. By
decreasing the number of chickens per squared meter, and increasing the distance between farms we can
already prevent more outbreaks, and make poultry lives more comfortable. We can have a look on how
our solution might work together with the aforementioned strategy.
Our Next Steps
Get into contact with new stakeholders which came up in the interview (e.g. experts in avian
influenza in wild birds and organizations which organize case reports of avian influenza in wild
birds).
Review the ‘National Action Plan to Strengthen Zoonoses Policy’ and possibly conduct an policy
analysis regarding avian influenza
Find experts in disease transmission and epidemiology regarding avian influenza
Aim to possibly create a model of avian influenza spread among wild birds and poultry, to
identify where to first implement our solution to prevent infection most optimally.
Date of meeting: 19-07-2022
Description
This stakeholder works in the poultry meat processing industry. This is the part of the sector which comes
after the farmers and the poultry parent companies. They only represent the interests of this part of the
sector. To gain their perspective, we conducted an interview to figure out how these enterprises get
impacted by avian influenza, and how they are involved in tackling this pandemic.
Contribution
The stakeholder told us that this part of the industry is most certainly affected by avian influenza. We
learned that meat processing companies also have restrictions if there is an avian influenza outbreak
nearby. However, this depends on how far it is located from the outbreak epicenter. On top of that, meat
processing companies can be impacted through gaps in their supply lines through culling’s, transportation
barriers and export restrictions. These measurements can all be a result of avian influenza outbreaks at
earlier stages of the supply chain. Later in the supply chain (e.g. meat processing companies) this can
result in deliveries delaying or not coming at all. Of course, for farmers this is an enormous economic
burden. However, companies up- and downstream of the supply chain have at least as much financial
damage. This costs their sector tens of millions of lost profit (mostly because of export restrictions). The
stakeholder emphasizes that the damage is not only in the farming sector, but there are definitely damages
surrounding the farmers which are influenced by avian influenza.
Stakeholder states that avian influenza outbreaks in the poultry industry most often originates from wild
birds. Still, through which routes it actually enters the industry is difficult to pinpoint. It could be through
dust, feces, direct contact, or other routes. Even though biosecurity measures are in full effect, there have
still been quite a lot of outbreaks. Therefore the sector is working hard to look at additional measures to
handle this disease. Stakeholder expresses that on top of hygiene and biosecurity there is also more and
more support towards vaccinating the animals.
Adjustments
Stakeholder identified a few things about vaccinations where we might be able to improve upon. One of
the problems with vaccinations is that you would need new variants every once in a while because of
mutations. Also, when a sub optimal vaccine is used, the virus could still infect vaccinated birds which
would then not show symptoms. The virus could then silently move through the bird population which is
unwanted.
We decided to slightly change our way of approaching and explaining our solution to stakeholders. To
our current believes our protection strategy should improve upon the aforementioned lacks in vaccination.
Therefore, we will focus on these parts whilst talking with stakeholders and gain more knowledge about
these aspects of our solution. Stakeholder underscores that a huge aspect in whether this solution could
ever be a success, is if it were to be more efficient, more effective, and save money.
Our Next Steps
We have discussed a lot of different stakeholders which we were not aware of before, therefore
we decided that we were going to create a stakeholder map which we can update after each
meeting if necessary. This way we can keep track of everyone involved, and what group we still
need to speak to.
Do a cost-benefit analysis: our solution vs. conventional vaccination (vs. current status-quo). For
this we will also have to consult stakeholders which we aim to find.
Date of meeting: 19-07-2022
Description
This poultry farmer started of with a large farm of around 5000 poultry. Now they have a smaller farm of
around 300 poultry. The poultry they have are kept for their meat and are held around 30 weeks before
slaughter. During this interview, we focused on the impact that the avian influenza pandemic has on
individual poultry farmers. Also, we were curious to the standing of them towards vaccination in general,
and our own solution.
Contribution
The stakeholder states that as long as you do not actually have an avian influenza outbreak in your own
farm, caging your poultry up as a preventive measure has the largest impact. This caging up would be a
result of an outbreak close to your own farm. This provides farmers with a variety of problems. Besides
not allowing visitors, which for small farms which also function as stores can be detrimental, there are
also other logistical issues. A good example of the logistical issues arising from caging up poultry is that
most farms are not build for keeping poultry inside for too long. This is a result of regulation requiring
poultry to free-range and have more living space. When a farmer has to cage up their chickens, these birds
have to live inside for quite some time which has both an impact on the farmers and chickens. The
farmers might not be able to sell poultry products as being from ‘free-ranging chickens’ anymore, since
they had to be kept inside for a prolonged amount of time. Besides that, chickens in small confined spaces
have a higher mortality rate. For example, because males cannot be held together due to aggression, but
must also be kept separate from females for similar reasons. The stakeholder states that they are definitely
afraid of the virus, and that we have to do something about these outbreaks quickly. They state that it is
nearly impossible to keep the virus outside of your poultry enterprise, therefore solutions such as
vaccination are needed. If it where possible, the stakeholder would immediately vaccinate their poultry.
For him trade regulations are not necessarily a problem, as he only sells locally.
Adjustments
The stakeholder says that our solutions seems feasible in bigger modern farms. However, they state that it
might be more difficult to implement for hobby keepers and small scale farms. We will have to think of a
way to make our solution feasible for these parties as well, or make sure that protecting this group of
poultry is not necessary in order to prevent large scale outbreaks. The stakeholder also expresses that we
have to be certain that the bacteria will actually colonize the lungs, rather than dying off a day/week later.
Also, when farmers have to spray their poultry multiple times our solution will not be accepted in the
poultry industry. Therefore, we will have to adjust our project in order to ensure colonization of the lungs
upon treatment.
Our Next Steps
Talk to more (large-scale) poultry farmers in order to find out their perspectives regarding the
avian influenza pandemic and on their stance towards our solution.
Talk to individual veterinarians about the effectiveness of our solution.
We have to think about a model to study and optimize bacterial growth in the lungs in order to
make our engineered bacteria survive there as long as possible.
Date of meeting: 20-07-2022
Description
This stakeholder is very involved in avian influenza. Through diagnostics, screening and sampling at
farms this stakeholder directly interacts with the avian influenza crisis in The Netherlands. The stakeholder
is also engaged in research and discussions concerning avian influenza with both the Dutch government
and international regulatory bodies. We conducted this interview to gain a broader perspective on the
avian influenza epidemic, from an overarching international (EU) perspective. This stakeholder is also
very well versed in poultry health/veterinary science, so we were also eager to hear their thoughts on our
project.
Contribution
Besides the highly pathogenic H5 variant, there are also lesser pathogenic strains such as H9N2 which
rage through the world. These variants are just as dangerous as H5 as potential zoonotic viruses.
However, they currently only spread in countries outside of the EU, which often do not have the means to
deal with such an epidemic. As these variants do not affect the EU directly and are less pathogenic for
poultry and wild birds, they are often left to rage through these countries unchecked. The stakeholder
states that creating vaccines for these variants, and making them available for the whole world (not just
first world countries), will be crucial in tackling this avian influenza epidemic.
The stakeholder talked to us about how it is very likely that avian influenza outbreaks in poultry are a
result of introductions from wild (migratory) birds from other countries. The last 2 years, the H5 strains
are even able to maintain in the local wild birds during summer. Therefore, even though our own
biosecurity and screening policies are very strong in The Netherlands, we are still unable to keep the virus
out completely.
Conventional vaccination works well as long as the wild virus stem resembles the vaccine strain enough
and birds respond well to the vaccination. For optimal efficacy in long living birds, you would have to
vaccinate several times. The stakeholder states that it is likely that we will have to work with a
combination of vaccines. A major disadvantage of the conventional vaccination is that it is very hard to
distinguish infected from vaccinated birds (DIVA principle) what is needed to be able to show that the
flocks are free of AI virus. The new generation vaccines (DIVA suitable) are likely much better, but we
have no data yet on whether they tackle the spreading of the disease more efficiently. The stakeholder
confirms that all of the vaccines for highly pathogenic avian influenza (including the next generation)
make use of the conventional ‘needle’. This is because other techniques like spraying do not evoke a
(proper) immune response to sufficiently protect poultry. The stakeholder states that these injection
vaccines are simply not the optimal solution as it is costly and labor intensive. They can therefore hardly
be applied in an emergency situation (vaccinate large flocks close to outbreaks). The stakeholder
expressed clear benefits that play into this, which our solution offers as well:
Potential to more effectively prevent spreading of the virus.
Mass-application vaccination (e.g. a spray) would be the optimal solution, and still does not exist for
highly pathogenic avian influenza.
Adjustments
This stakeholder especially made us realize how other countries look towards this epidemic. Mostly the
stance of non-western countries towards avian influenza. At his recommendation, we started looking into
this perspective as well as how our own project could play a role in this. The stakeholder also underscored
the fact that there will not be one single solution to this epidemic. A viable solution would most likely
still co-exist together with other measurements like biosecurity or a combinations of vaccines. This made
us set out to figure out how our strategy can co-exist in the framework that is already in place/ will be in
place to tackle avian influenza. The stakeholder expressed concerns about only tackling the lungs of
poultry, as the virus might enter them through other pathways as well. After the interview, we adjusted
our plan and we set out to make sure our ‘nanobuddies’ can colonize the gastro-intestinal system as well
for better coverage. The stakeholder also expresses that if you are able to effectively and easily protect
against a non-pathogenic variant, it would still be a breakthrough. Therefore, we adjusted our project so to
also research the binding efficacy against other AI variants as well.
Our Next Steps
Find non-western countries (industry or governmental) to talk to about their stance on avian
influenza.
Talk to experts in other countries provided by the present stakeholder.
We realized that it would be very interesting to look at the binding efficiency of the nanobodies
produced by our GMO to different viral strains after we conducted this interview. We have
gathered a lot of different hemagglutinin proteins (e.g. H3, H5, and H9) to test the binding
efficiency of our nanobodies.
Date of meeting: 21-07-2022
Description
We have conducted several interviews about the origins of avian influenza where wild bird populations
came up. Therefore we set out to speak to people that are on the front line of tackling avian influenza in
wild birds. The animal ambulance is involved in gathering diseased animals and treating them. This
includes the gathering, and often culling, of avian influenza infected birds. We were welcome to visit one
of their locations and speak to them about the impact this epidemic has on their day-to-day, and their
perspectives on the crisis itself.
Contribution
The stakeholder expressed that the avian influenza epidemic certainly plays a big role in their
organization. The past few years there has been an increase in cases of avian influenza in wild birds for
which they had to respond. The length of flu seasons also increases by the year. The increasing impact of
avian influenza has a big impact on the animal ambulance, and their employees/volunteers. It is not a
pretty sight to see a bird that has been infected by this virus. On top of that, there is also an increasing
amount of costs they make which has a big financial impact. For example they had to invest in protective
suits, disinfection materials, and another vehicle. This epidemic definitely puts pressure on the
organization both logistically and financially, but also emotionally. Employees dislike going to avian
influenza cases because of the suffering the animals often are in, and their disability to really help these
animals. There is also an increase on the workload for veterinarians.
When we entered the location the first thing we noticed immediately was a sign on the door reading
‘XXX’, and a disinfection mat before the door. This emphasizes more extensively on how even these
organizations are heavily affected by the epidemic, and have to constantly be aware to not further spread
the virus.
The stakeholder explicitly stated that vaccination in wild animals will not be an option, and that this
epidemic would have to be tackled elsewhere.
Adjustments
For this interview we did not necessarily discuss our solution explicitly. However, the stakeholder told us
that as long as the solution is deemed safe, they would be prepared to eat GMO treated meat. They did tell
us that terms like ‘nano’ and ‘GMO’ could be bad if you would like to sell your solution to a large public.
These terms may work as a repellent as they could come over as ‘scary’ or simply difficult to understand.
We decided to find ways of explaining these terms to a more general public so they become easier to
understand, and therefore potentially less ‘scary’.
Our Next Steps
Contact the stakeholders suggested by the present stakeholder to further investigate.
Find out why collection, diagnostics, and culling of wild infected animals is not centrally
orchestrated by an (governmental) institution. Especially since this is done for some other
diseases.
Date of meeting: 22-07-2022
Description
This stakeholder is a practicing veterinarian specialized in poultry. After previous interviews with other
stakeholders, we set out to also get the perspective of a veterinarian which is practicing in the field on the
Dutch avian influenza crisis. We also hoped to gain some insights on what they thought of the real-world
practicality of our proposed solution.
Contribution
The stakeholder is involved in the poultry industry by providing consults on biosecurity and improving
poultry health, as well as diagnosing and early prevention of avian influenza in poultry. They also state
that awareness among poultry farmers is of great importance in tackling this epidemic. For example,
veterinarians assist in providing knowledge on red flags, and how to detect avian influenza early on in
poultry flock. According this stakeholder the biosecurity is already of high level in The Netherlands. On
top of that, the protocols in place for early outbreak management also seem to be close to perfected.
However, monitoring of avian influenza in poultry farms could be improved upon. This stakeholder also
provided us with a substantial number of stakeholders we were previously not aware of. The stakeholder
stated that a flaw of conventional vaccination is that these vaccines especially work well in treating
symptoms of the disease. However, it does not excel in preventing actual transmission; vaccinated
animals can still harbor the virus, and transmit it while having limited symptoms themselves. The
stakeholder expressed that this could definitely be a good selling point of our proposed solution, and it
should therefore definitely be something we should research extensively. Also the possibility of our
solution to work quickly could be an improvement upon regular vaccines. The stakeholder states that
these often take 5 weeks offer full protection against the virus, which is suboptimal in the poultry
industry. Lastly, this stakeholder also states that it is highly unlikely that there will be one single
solution/vaccine which will solve this epidemic. It will most likely be a combination of measurements
that enables us to effectively keep the virus out of poultry farms.
Adjustments
The stakeholder expanded our knowledge on stakeholders and we adjusted it accordingly for our
stakeholder analysis. They also made us aware of calculations carried out by AVINED with regards to the
impact of the avian influenza epidemic. We plan to implement this data in our literature review on the
avian influenza crisis. The stakeholder also had an interesting comment on our delivery method. They
suggested that we could use a food supplement to also ensure colonization in the gut, a place where avian
influenza can also infect poultry. Therefore, we aim to talk to companies providing food supplements
(including probiotics) to poultry businesses to hear their thoughts and feedback.
Even though the stakeholder themselves would be fine with eating meat from poultry treated with our
GMO probiotics, he is worried that the general public (especially those without a background in biology)
might not be ready to consume meat treated with such strategies. After hearing this from multiple
stakeholders this was the straw that broke the camel's back, after which we decided to conduct a
questionnaire on the avian influenza crisis, GMO’s in the meat industry, and our project.
Our Next Steps
Design a questionnaire, look into the ethical considerations, and get into contact with the ethics
committee for social sciences from the University of Groningen.
Update our stakeholder map, and get into contact with newly provided stakeholders. This
includes finding food supplement companies which are able to speak to us about our solution.
Date of meeting: 25-07-2022
Description
As a result of previous interviews, we wanted to gain the perspective of other stakeholders from other
countries. We also aimed to speak to more individual poultry farmers. In this interview, we tried to
combine these two goals, and we spoke to a stakeholder who is involved in farming poultry in Germany.
We were primarily interested in how the avian influenza epidemic impacts other countries, and how they
deal with this crisis.
Contribution
This stakeholder in involved in a family owned poultry farming business spanning across multiple
locations. They describe that the systems in place to prevent and anticipate avian influenza are similar to
The Netherlands, which makes sense as these countries are both under EU regulation. These also include
biosecurity, monitoring, caging, and culling for example. They also think that the outbreaks start as a
result of wild birds infecting the poultry, similarly to how Dutch stakeholders think the outbreaks
originate. They also state that poultry farms in Germany are all densely located in specific areas of the
country, which makes avian influenza outbreaks harder to contain. The stakeholder describes that avian
influenza is certainly something which keeps the German poultry industry busy. They themselves have
also had occasions where they had to cull all of their poultry stock as a result of an avian influenza
outbreak. This has a big economical impact on everyone involved in the poultry industry: from primary
breeding companies to retail. The stakeholder describes how free-ranging policies also become more
difficult to uphold due to the ever-growing presence and spread of avian influenza. We asked the
stakeholder where they thought a vaccine or our solution would have the largest impact. They stated that
outbreaks might have a bigger impact on the part of the industry working with parent and grandparent
stocks. Lastly, we talked about how high avian influenza is on the agenda of German authorities. Even
though the problem is becoming increasingly more urgent, German regulatory authorities are not very
keen on changing the status-quo. However, internal talks within the poultry industry about tackling the
avian influenza crisis are increasing. Since public opinion towards the culling and caging of chickens is
also increasingly worsening, the stakeholder expresses that they have faith that the relevant parties will
soon come together to find a solution.
Adjustments
Similarly to the other stakeholders we spoke to, this stakeholder states that the biggest bottleneck of the
implement ability of our solution would be public perception. This further stimulated us to develop our
questionnaire, and find ways of gathering information from both Dutch, and international general public.
The stakeholder also underscores that it will be crucial that a solution would cut costs for poultry farmers
themselves. Since preventing and monitoring avian influenza outbreaks is mostly the (financial)
responsibility of the farmers themselves, whether they will use our solution will heavily depend on
whether it saves them money. As a result of the interview, we also set out to speak to Dutch authorities to
talk about where implementation of our solution would have the most impact (e.g. primary breeding stock
vs. broiler farms).
Our Next Steps
Find out on which section of the poultry industry to implement our solution, in order to have the
largest impact. This could have implications on our delivery strategy.
Gain insights on the ethics and policy around GMO’s in the food industry. We plan on doing this
through a collaboration with other iGEM teams working on GMO’s in a big stakeholder meeting.
Make an English version of the questionnaire to also target an international audience.
Date of meeting: 26-07-2022
Description
We set out to speak to an expert in the field of transmission of avian influenza. This stakeholder works as
an epidemiologist among others on avian influenza. Besides assisting and investigating poultry farms
during potential outbreaks, he is also involved in research into the origins of outbreaks in poultry.
Through this interview we hoped to confirm certain information which we heard in various interviews
regarding vaccination and transmission of the disease. This stakeholder also has experience with working
with disease-transmission models, so we hoped to ask him some questions about this as well to potentially
include it in our project.
Contribution
Through observing and tackling samples from farms which have suffered outbreaks of the virus the
stakeholder analyses where the origins of the virus lies, and which variants are dominant. The stakeholder
confirms that we know with some certainty that the virus gets transported through wild birds from Asia to
the EU via common breeding grounds in Siberia. In Asia, the virus is endemically present in poultry due
to different styles of keeping poultry, more loose regulations, and less preventive measurements. Wild
birds get infected in Asia by spill over of the virus from poultry. Infected wild birds from Asia migrate to
common breeding grounds in Siberia and come into contact with migrating wild birds originating from
Europe. With the autumn migration, these wild birds of European origin come back to Europe, taking
with them the virus they received in the common breeding grounds in Siberia. Still, it is unclear how the
virus actually travels from contaminated areas in the neighborhood of poultry farms in The Netherlands
into poultry units according to the stakeholder. During the bird flu season, all outdoor poultry are housed
by obligation inside the poultry buildings, preventing contact between potentially infected wild birds and
poultry. This is especially a mystery since biosecurity measurements are quite extensive in the
Netherlands. We further discussed some of the hypotheses regarding poultry farm infections. It is unlikely
that the virus travels into farms through air transport of contaminated feces from infected wild birds into
poultry buildings via ventilation openings or via aerosols of breathing air from infected wild birds via
ventilation opening in the poultry units. However, research is still being carried out to rule these pathways
of transmissions out. It has been proven that the virus remains present and survives in cold water for a
long time during the winter periods, which could be a possible way of poultry getting infected if they
drink from this water. This appears to only go for low pathogenic avian influenza during other periods of
the year however because keeping poultry outdoors is forbidden during the bird flu season. The most
likely way the virus gets in is through biosecurity standards not being properly met, leading to outbreaks
in poultry farms. According to the stakeholders study from other countries, there are indications of flaws
in biosecurity being possible, and it is reasonable to believe this would also happen in The Netherlands as
well. Vaccination is likely necessary to address this crisis. However, vaccines often take long to develop
and there are a lot of virus variants raging over the world. This would mean you need to have a lot of
vaccines ready to go, and constantly monitor which variant is dominant. The stakeholder states that for
avian influenza this will become a very tedious process. Also, the stakeholder states that the actual
administration of the vaccine is also a big bottleneck. On top of that, these conventional vaccines mainly
focus on clinical protection, rather than prevention of transmission.
Adjustments
After the interview we realized that even though our nanobodies are broadly neutralizing, it will still be
crucial for the system to be very dynamic to also be useful in the long-term. We strived to adjust our
project in such a way that it will become easy to switch between different nanobodies which can target
different specific virus strains. The stakeholder also provided us with a lot of information about animal
disease transmission models. The stakeholder shared the basics of these models with us and inspired us to
find ways on how we could integrate these models in our project.
Our Next Steps
Estimate how long the GMOs will stay in the lungs. As this will greatly influence the actual
implement ability of our solution. We aim to do this through reviewing literature and possibly
modelling.
Get into contact with companies which provide spray vaccinations to ask them whether we can
use one of their vaporizer to research and predict how efficiently the virus actually enters poultry
lungs.
Look into the possibility to integrate transmission models talked about with the stakeholder (SRI).
August
Date of meeting: 03-08-2022
Description
We organized a meeting with a biosecurity officer from the animal welfare institute (IVD) from the
University of Groningen. We mainly did this for two reasons, namely, 1) find out the Dutch regulations
regarding the use of animal by-products for research purposes, and 2) to get feedback on our
biocontainment strategy.
Contribution
The stakeholder quickly put us on the right track to conduct our proposed research plan by informing us
of the Dutch Animal by-products Regulation. They also provided us with documents extensively
describing the people we would need to contact and steps we need to take to make our experiment a
reality. Besides this, they also put us in contact with someone with experience in contacting commercial
meat processors for animal by-products for educational or research purposes. We also discussed how we
can set up the experiment, taking into account the limitations of the Dutch regulation. This also includes
insurances of high quality sampling of the by-product, location to conduct the experiment, and a location
to get rid of our waste (a risk 1 waste material).
We also discussed the kill-switch design we have thus far, and talked about what are important steps in
biocontainment. The stakeholder explained the principles behind biocontainment, and we discussed how
we might add on or improve upon our current kill-switch strategy. As making our cells dependent on
metabolites not naturally found in the environment would for example be not suitable for our strategy, we
discussed alternatives such as making a bacteria dependent on a particular symbiosis with another
Lactobacillus species.
Adjustments
We adjusted the approach of the microbiome analysis to better fit the legal framework of The Netherlands,
as well as the ethical framework regarding animal by-products as defined by the iGEM HQ.
Our Next Steps
Contact iGEM HQ again to find out what sort of experiment will fall under the animal by-product
exemptions.
Contact a microbiome analysis company who can do our analysis.
Contact a poultry meat processor / butcher who can provide us with lungs (by-products of meat
industry).
Conduct our lung microbiome analysis.
Date of meeting: 11-08-2022
Description
Our good friends over in Freiburg, Germany, notified us about this scientific integrity workshop they
have been organizing. We had met them at the European meet-up and been pen pals ever since, so it was
not a difficult decision to participate in their workshop. They had invited Michele Garfinkel, head of the
European Molecular Biology Organization (EMBO) policy program, who held a talk about scientific
integrity, and how to present results from your research properly.
Contribution
During the workshop we learned a lot about the pitfalls that scientists often fall in while presenting their
research. For example, how cutting off images could lead to misleading results. Michele also showed
examples of how lighting on images might lead to a researcher misinterpreting their results, unknowingly
drawing false conclusions. The presentation also included a funny example of a scientific article that their
organization had to review about plants. When showing the different plants in the article, the organization
discovered that every image containing different types of plants showed the same environment (such as
nearby rocks and twigs which are exactly aligned). They thought the author had fabricated the results (the
plants) as it appeared it was just the same plant at different timepoints. However, after consulting the
author about this mystery, it appeared that it is a common practice to place all of your plants in the same
ground before making the final picture. The only reason for this being that the ground would then be
neutral and more pretty than the ground the plant originally grows in. This example is a good example of
how you should not always assume the worst, but also be very clear on how you obtained your pictures.
All in all, we learned a lot about data visualization and properly showcasing results.
Adjustments
Besides all of the interesting ways to present data we had discovered which we could learn, we decided to
review how we are presenting the results we had created thus far. Could they for example be misleading?
Moving forward, we will also take extra consideration about scientific integrity during our iGEM project.
For example, not over-stating the actual potential of our project and outlining its flaws to stakeholders as
well. This could also prove useful to us, as this opens us up to even more feedback from experts and
stakeholders.
Our Next Steps
Review the visualizations of the data we have collected so far.
Critically review our literature to see what the potential impact of our project could be. Also
realistically review whether others using a similar approach have succeeded, or where they failed.
Date of meeting: 11-08-2022
Description
For our project we thought of several educational outreach activities we could organize. We mainly
wanted to organize an activity (e.g. small experiments in the light of our project / synthetic biology) at a
Dutch high school or pre-school. After coming up with some ideas we got in contact with someone that
works in science communication for feedback on our ideas and our plan of approach. We also used this
opportunity to go over our project and discuss ways to efficiently explain our solution to the general
public.
Contribution
First we discussed ways we can actually approach schools to participate in our educational activity. Then
we talked about how we can make our activity as useful for the students as possible. We both want them
to get excited about synthetic biology, but also give them knowledge they can actually use in their studies
(e.g. high school biology final examination). We discussed this with our expert and they told us that it
would be wise to check the curriculum of the high school biology course, and structure our activity
around the learning goals stated there. This could not only make our activity a nice addition/expansion to
their curriculum, but also makes our activity more appealing to busy high school teachers. After this we
talked about how to structure our activities to make them as interesting/engaging as possible. It is also
important to not make it too childish.
Adjustments
We learned a lot about how to organize these types of activities and we now have the framework we want
to use to create our activities. We came up with the idea to do an escape room in the theme of ‘escape the
lab’. Through puzzles and problem based learning we want to bring the terms in students’ curriculum ‘to
life’. The most important thing we learned is that it is very important to consider the fact that high schools
are busy and that extracurricular activities need to be well structured and planned before they would
consider doing it. We created a very comprehensive ‘workshop description’ which explicitly states the
learning outcomes and topics discussed in the activity.
Our Next Steps
Fine tune our workshop description page and start contacting high schools.
Design an escape room for young students which aims to strengthen knowledge about synthetic
biology in the light of our project.
Date of meeting: 15-08-2022
Description
This stakeholder is occupied with a variety of wild and domestic animals which are in danger. This
stakeholder got into contact with avian influenza about three years ago, before that the topic was less
relevant in general, and to the organization. We wanted to speak to this stakeholder to gain information on
the perspectives from hobbyist poultry keepers and wild/domestic bird welfare advocates.
Contribution
The stakeholder first started off with explaining the huge differences between avian influenza policy for
wild/domestic birds and birds in the poultry industry. To put it bluntly, there are about two hundred full
pages on policy regarding outbreak prevention and measures in farms, while there are only a few pages
available outlining policy regarding wild birds. Simply said, wild animal welfare advocates such as the
animal ambulance and other NGO’s are left to come up with policies themselves, leading to a variety of
issues and human error. The stakeholder, together with the organisation they represent, is working on
providing this policy and getting it to the right people. For organizations trying to safe wild birds avian
influenza proves a logistical and economical challenge (e.g. the need for special quarantine rooms). It is
also difficult to discriminate between sick birds without avian influenza. The stakeholder underscores that
there is a need for quick ‘self-tests’ which can be easily deployed to find out whether an animal has avian
influenza. At first we thought it might be a good development that a single (governmental) organization
would take the lead in tackling avian influenza in wild bird populations. However, after speaking to this
stakeholder we learned that most organizations that would have to do this are already heavily over
capacity with avian influenza in the poultry industry. It would be better to invest more and to improve
upon already existing infrastructure and policy.
Adjustments
The stakeholder provided us with a lot of material, especially regarding the policy side of avian influenza.
They also prompted us to look into debates in the Dutch government about avian influenza. All this
information can be put into a policy analysis. On top of that, the stakeholder provided us with a lot of
interesting stakeholders which we have not yet spoken to. We also spoke about implementing protection
strategies in wild bird populations, which we quickly found to be almost impossible. The stakeholder did
give us some tips on the implementation of our project. They advised us to implement a pilot in a small
farm or petting zoo, or even a wildlife rehabilitation center, to mimic the conditions on a regular poultry
farm best; there are vast differences between poultry in a research setting and a farm setting. The
stakeholder also explained to us the importance of ‘selling’ our product to the general public. We talked
about ways to explain our project to the public in a way that does not downplay implications from our
solution, but also showcases the good it could do.
Our Next Steps
Look into the possibility to (conceptually) design a virus detection platform using nanobodies.
Adjust our future implementation plan.
Study all the policy documents and governmental debates regarding avian influenza
Date of meeting: 08-18-2022
Description
This stakeholder is very involved in combatting avian influenza in the poultry industry and often works
together with worldwide organizations that preach for animal welfare. We wanted to speak to this
stakeholder as they represent the main authority in The Netherlands that regulate food and consumer
product safety. This organization is also the main authority on avian influenza and disease policy in the
poultry industry. A lot of stakeholders in the past also advised us to speak to this stakeholder about avian
influenza and our solution.
Contribution
We spoke to the stakeholder about the impact this epidemic has. Besides the economical impact, there is
also a heavy toll on the organizations that have to deal with the outbreaks. People working for these
organizations have to work very hard to deal with and prevent outbreaks in farms. The stakeholder also
took us through all the steps that are taken (mainly by the NVWA) in order to keep the disease out of
farms, which also includes outbreak management protocols. They pointed us to the protocols which are
available through their websites.
We also talked about the development of vaccines, and how they could help combatting this pandemic.
Developing vaccines takes a long time, and dominant serotypes of the virus often switch rather quickly.
There is a desperate need for a broadly targeting vaccines. Vaccination should also be automated per
injection in case of live attenuated vaccines, or be able to be sprayed in case of dead vaccines. Otherwise
the implementation of these strategies will simply not be durable and feasible. Of course, from the
perspective of our project, this confirmed that these are important factors to focus on in the
implementation of our own protection strategy. Personally the stakeholder thinks that we should
definitely vaccinate our poultry. However, the World Organisation for Animal Health (WOAH, former
OIE), which consists of 188 countries, all will have to agree with each other on this difficult decision as
well. These countries are all mainly concerned with how this will affect trade. Discriminating between
vaccinated and infected meat for example is a big issue in the light of trade. Our solution would also have
an edge over conventional vaccination on this front.
Adjusments
The stakeholder specifically stated that bird feces are very contagious and can spread disease very easily
e.g. transport vehicles, materials, professional farm visitors etc. We have to take this into account while
designing the final plan of our solution. Especially since we are working with a microbe that is able to
habituate in the gut microbiome of chickens as well, we will adjust our plan to also incorporate the gut
microbiome colonization of our Nanobuddies.
We also discussed where our solution would have the biggest impact. Since we would not be able to
immediately vaccinate every bird in the Dutch poultry industry, we need to know where we would have
the biggest impact. After discussing it with the stakeholder, it came down to a top-to-bottom approach.
The birds that need protection the most are the ones on top of the so called ‘poultry-pyramid’. These are
the grandparent and parent breeding stocks which provide flocks of poultry to the entirety of the poultry
industry. If these would deal with less outbreaks, the whole industry can benefit. Besides using this
information to adjust our future implementation strategy, we also keep this information in the back of our
minds while taking the last designing steps for the solution itself.
Lastly, and most importantly, the stakeholder talked about how vaccination not necessarily prevents
disease spread. Finding a way to especially prevent disease spread is crucial and is something the industry
really needs (besides any vaccination strategy).
Our Next Steps
Have a look at the protocols and operational manuals on the NVWA website and contingency
plans on the website of the Ministry of Agriculture, Nature and Food Quality (LNV).
Adjust our future implementation strategy.
Further research the rate of disease spread after vaccination and our own strategy.
Date of meeting: 23-08-2022
Description
As most of the stakeholders explained to us that a lot of the policy relevant to our solution is centrally
regulated by the EU, we set out to speak to experts all around Europe. This stakeholder feeds information
to European organizations of veterinarians with regards to the poultry industry. They also advice their
government on avian influenza strategy and biosecurity. The aim of this interview was to gain the
perspective of non-Dutch stakeholders on avian influenza and our solution.
Contribution
We quickly learned that all of the regulation and measurements against influenza are the same in the EU.
There are only slight differences regarding avian influenza in wild bird populations and in hobbyist
poultry keepers. These hobbyist poultry keepers also might be a big contributor to outbreaks in larger
scale poultry business. Protecting these groups of poultry is therefore very important. We compared
policies from both our own countries and quickly found that the issues are quite similar. The stakeholder
also states that there are still a lot of improvements that could be made in biosecurity and early warning
systems in all of Europe. As of know, there is still quite some room for human error which might
contribute to disease spread. Countries such as The Netherlands and France are pushing hard on
implementing vaccinations. However, getting vaccinated does not necessarily mean that the birds can not
get the disease. The stakeholder also speaks of a vaccination pilot in Italy which did not go down that
well. Still, vaccination looks like the only solution as biosecurity measurements prove insufficient. The
stakeholder explained to us that a problem with vaccination is that it does not necessarily protect against
spread of the disease; a common theme about vaccination in a lot of our interviews thus far. The
stakeholder also expressed a need to quickly and easily diagnose avian influenza by means of a self-test in
almost every country in Europe.
Adjustments
The stakeholder told us that for our strategy we would have to take into account a few key concepts which
vaccines sometimes lack. This includes protection against infection (preventing disease spread),
deploying strategies (logistics), and whether it works on different types of poultry. This last part is
especially relevant to us since we have not yet considered how and if our Nanobuddies can also live in
geese for example. Even though chickens are by far the largest proportion of livestock, we have to
consider that we can also protect other types of poultry.
The stakeholder also helped us think about how our solution would fare in the real world together with all
other systems which are already in place. For example, even after implementing our solution, the poultry
industry would still need constant monitoring for avian influenza.
The stakeholder also gave us interesting ways and ideas to test whether our solution will work for our
future implementation plan. They also underscore that there will never be a ‘silver bullet’, and that every
solution will have to co-exist in the industry to achieve the highest effect.
Our Next Steps
There appears to be a clear need for diagnostic tools which can easily diagnose avian influenza
(much like a COVID self-test). Even though it is already quite late, we aim to find a iGEM team
which we could collaborate/ partner up with to create such a thing.
Implement the suggestions of the stakeholder for our future implementation.
Date of meeting: 26-08-2022
Description
The second meeting with the RIVM was intended to show how our project was progressing, especially
with regards to safe-by-design we have implemented along the way. We showed our progress and our by
now more elaborated solution design. This was a good opportunity to get feedback from experts that have
already seen where our project started, and are more involved in the specific design choices we have
created.
Contribution
After our presentation, we got to discussing how we might be able to improve upon our design in terms of
safety, a very important key-value in our project. We discussed several aspects and after a few minutes of
discussion we often came to the conclusion that we had actually thought of it already, and made changes
to the design accordingly. The RIVM was very impressed by our kill-switch design. As a result of the last
meeting, we have created a 2 input OR-gated kill-switch which is sensitive to light and temperature. We
discussed how we could maybe even improve further on this design. This mainly touched upon actions
like genomic integration to improve genetic stability of our constructs across generations of bacteria. We
have only considered using plasmids up until now, so this was definitely an interesting comment on our
design. Still, we have decided that this specific genomic integration would be outside of the scope of our
research, as we already had enough trouble transforming our bacteria for the experiments we have already
planned.
The RIVM brought us to the attention of the fact that before the animals will be transported to be
processed for slaughter, you would ideally not have the GMOs still (alive) in the lungs. They asked us
whether we had thought about this, to which the answer was ‘no’. We simply assumed that when a
chicken would be slaughtered and killed, body temperature would decrease thereby activating the
killswitch. However, this increases the risks of our GMO escaping to the world when they travel through
a lot of different environments; potentially finding an environment where they could somehow thrive. It
would also be nice to have a way to ensure that our GMO is 100% eliminated from the animal if the
situation calls for it, a sort of fail-safe regardless of temperature and light. During the meeting, an astute
team member remembered something about our kill-switch which we have not utilized for the solution we
have created.
Adjustments
After speaking to the RIVM we got to thinking, and realized that there might be a property which is
already designed into our kill-switch. This is very interesting and kind of random as it is just something
we read about when designing our killswitch and reviewing literature. Now, we realized that we could use
certain properties of our kill-switch that we did not originally intend on using. However, one of the parts
of our kill-switch can be activated through a non-toxic chemical which makes our kill-switch essentially a
3 input system. When checking our designs, and making sure that the specific pathway the non-toxic
chemical interacts with still works as described in the literature, we concluded that there would be no
reason why our kill-switch can not be activated through the deadman kill-switch system! We are unsure
how we could test this in the lab considering we still have a long way to go before we can test the
previously designed kill-switch, let alone this particular feature. Still, we will include this in our
implementation and think of how we can utilize this new feature to improve the safety, but also the
usability of our solution!
Our Next Steps
Further research and try to incorporate the third input to our kill-switch.
Thinking of how this new kill-switch design can be more useful, and weighing whether it would
be good to allocate resources into further developing this specific kill-switch as a whole.
Design experiments to potentially test this newly found input.
Figure 4. Online meeting with RIVM. Present were: Mink, Bindert, Sander, and Isis.
September
Date of meeting: 09-09-2022
Description
This stakeholder is active in improving the animal welfare of birds in the poultry industry through policy,
with a background in animal health and sciences. We wanted to conduct this interview as this specific
stakeholder was suggested to us by several of our previous interviewees. We aimed to use this interview
to find out important considerations regarding animal welfare, to gain a different perspective on avian
influenza outbreaks in the poultry industry, and for feedback on the future implementation plan for our
strategy.
Contribution
The stakeholder and their organization is very busy with keeping avian influenza out of the poultry farms.
One of the reasons this is important because this endangers the free-range industry, which is the future of
the industry where the organization is trying to build towards. An interesting thing we learned, is that a
large impact of avian influenza, besides the (preventive) cullings, is the downgrade of the egg marks from
grade 1 (free range) to 2 (barn egg) as a result of keeping the birds indoors. When chickens have to be
kept indoors for longer periods of time to decrease the risk of infection and farm-to-farm disease spread,
this downgrades their quality of life. Besides the impact this has on the chickens, this also massively
economically impacts farmers. These farmers made big investments in their free-range farms and the
quality of life for their animals, only to sell them for the same, or even less, amounts of money.
We spoke about the vaccines that are available already, and how these are not usable for the application
purposes we would have for them in The Netherlands. A lot of research is going into the development of a
vaccine that can be used. Until recently, there were little efforts in developing one. Most likely, this was
because the avian influenza epidemics where not as severe as they are now. No matter what bio-security
measures you install, you will never be able to completely keep out the virus. Especially since the best
type of bio-security measures often involve enclosing the chickens, which directly goes against the goal
of improved quality of life and more free-range chickens. Therefore stakeholders are all on the same page
with regards to the need of a vaccine. This organization, as well as many others, have to find a good
trade-off between protecting chickens against disease by means of vaccination and biosecurity, and
quality of life improvements by means of free-ranging for example.
We also discussed, as we did with many stakeholders by now, the roadblocks that still exist which prevent
the deployment of vaccination strategies. These of course mainly include the trade regulations, more
specifically the EU animal health legislation which prevents vaccinated birds from being transported to
other member states, which completely block out the use of vaccines for highly pathogenic avian
influenza. As far as the stakeholder is aware, the general trade regulations are not the main issue, but the
bilateral trade agreements could exclude trade in vaccinated animals and/or their products. The
stakeholder underscores that it will be very important to have new regulation implemented before we
actually have a new vaccine. Otherwise the change in policy might come in too late and the vaccine could
become outdated. However, there is a problem with that, as policy is only more likely to change as soon
as there is a vaccine ready.
There are a lot caveats with regards to our solution. Besides the upsides that the stakeholder recognized
such as: not dependent on the host immune system, and not using viral particles or live vectors, there are
still a lot of important things we will definitely have to think about. Mainly, the use of GMO’s in the food
industry and in live animals is a concern which is especially relevant to this specific stakeholder. The
stakeholder expressively said that it will be important that this needs to be safe for the chicken, as well as
for the environment. First the stakeholder was under the impression that we might genetically modify
chickens themselves, which raised a few eyebrows since that is something which is highly frowned upon.
After explaining and discussing our strategy, we also talked about how there is also more types of poultry
besides chickens which also need to be protected.
Adjustments
First of all, we did not yet consider the other types of poultry which are also affected by the spread of the
virus. The solution we designed is mainly directed towards chickens, which by far make up the largest
proportion of livestock. To make our solution more responsible and inclusive, we will also includes these
type of poultry. If we do not include these groups of livestock in the development of our solution, the
virus might just continue spreading; although probably at a slower rate.
As there was also some confusion about whether our solution was an ‘alternative to vaccination’ or an
‘alternative vaccination strategy’, we decided to review all of the abstracts, project descriptions, and wiki
texts. We for example already changed the abstract we send when contacting stakeholders to better reflect
what we want to achieve through our solution. We will also use this moment to clarify that we will
engineer a lung microbe, rather than the chicken/virus.
Lastly, the most important adjustment we have to take with us in our solution is the location where we
will implement our solution. The stakeholder told us that we should refrain from focusing on specific
groups of poultry (e.g. broiler chickens or grandparent stock). The only difference is the monetary value
of these poultry, which is and should not be what our solution is mainly about. Rather, we should focus
on the areas that are most often greatly affected. The stakeholder does mention that this is a choise, and
we could perhaps also put our focus on the parent stock. ‘Perhaps this might also lead to some form of
innate immunity in the offspring?’ Interestingly, in The Netherlands we have a lot of poultry farms around
nature reserves with a lot of water (with all of the water birds which often carry the disease throughout the
country). A more durable solution would be to relocate these farms, as well as keep farm density as low
as possible. The stakeholder recognizes that this will take time, and shorter term measures are also
needed. Therefore we need a way to quickly protect these birds specifically. The question of which birds
should be protected first and most often is not about value, but more about the risk they have of getting
infected, which could be related to geological location, type of farm, type of animals, etc. This is
especially important because that might mean we are able to keep these birds outside with better living
conditions, without the risk of the virus spreading at rapid rates, even though they live in a poultry dense
area close to nature water reserves.
Our Next Steps
Conduct a literature review on whether the bacteria also naturally lives in the lungs of other types
of poultry.
Implement the advice which the stakeholder gave about areas which are affected most frequently
and harshly, in the light of our future implementation plan.
Date of meeting: 15-09-2022
Description
As a tradition, the iGEM team of the University of Groningen is invited to speak at the GBB symposium
organized by the GBB faculty members. As iGEM is organized by the biomolecular sciences master,
which is a part of GBB, we got the opportunity to share what we have been researching for the past
months. For the iGEM team this is also a nice opportunity to see how it feels to attend a real scientific
symposium, and interact with the researchers doing very interesting research. We were hoping that some
faculty members might have some useful tips for us that we could use to improve our project.
Contribution
Before we got the opportunity to present ourselves, we looked at all the other researchers sharing their
posters and presentations about their research. Then it was the moment for us to present our research thus
far. Although we did not have a lot of dazzling results to share, we used this moment to share with
everyone how we were trying to achieve what we set out to do: find a solution to the avian influenza
crisis. We shared our delivery platform and experiment set-up to come to a proof of concept. After that,
we received a few questions regarding our project. Mainly, we discussed our biocontainment strategy
which we have extensively worked on for a while now so addressing that was not a problem.
After the presentation there was a session of informal drinks where you got the ability to speak to
everyone that was present at the symposium. We got to know dr. Max Fürst, an assistant professor who
approached us after hearing our plans for the modeling part of the project. He specializes in
computational protein design and high-throughput protein engineering; it is as if this meeting was just
meant to be. We had been stuck and struggling for a while now with regards to our nanobody model.
After a few drinks and a long conversation about our approach, he figured that he could help us try to
make it work. Although, he also mentioned that we are very ambitious with our plans and should take into
account that it is very difficult to create the model that we had envisioned. This could also explain why
we had been stuck for such a long time.
Adjustments
Moving forward, we have planned a meeting to thoroughly discuss our model and how we might be able
to improve on it. The conversation itself already yielded some good insights which helped us further a
few steps. Our plan now is to get a few critical aspects of our model working, rather than the whole thing
at once. This way we are able to produce some results as we are building our program. On top of that, we
also gained some feedback on our experimental designs. We got the contact information of a few people
that might be able to help us out with the specifics!
Our Next Steps
Have a meeting with Max, and discuss the plans we have for our modeling project.
Date of meeting: 16-09-2022
Description
We set out to speak with stakeholders that operate on a European/worldwide scale to gain their
perspective on avian influenza and our project. We spoke with someone from the IPC, who represent the
interests of the complete poultry industry value chain, even including companies providing machinery and
other services. The organization does not engage with governmental bodies and they do not try to
influence decisions or legislatory processes at national level (IPC provides inputs to OIE, Codex and other
international standard setting bodies). The stakeholder tries to create a platform for their members to
present their issues.
Contribution
We learned that this stakeholder is in close contact with the World Organisation for Animal Health
(WOAH) about issues such as avian influenza. The official position on avian influenza is that it is
definitely a big problem, and that it should be dealt with while not risking trade and food safety. They try
to facilitate conversation between their members and other stakeholders. Their mission is to create an
environment where animal health and welfare can be improved upon, without worrying about trade
regulations. They have a lot of knowledge in their organization, and they use this knowledge to build a
strong political message out of all of their combined experiences. We also learned that avian influenza
policy and measures are almost universal around the world although with some differences across
countries, as the large majority of countries developed their national legislation basing it on standard
established by the WOAH. For us this confirms that the problems we face in The Netherlands and the EU
regarding combatting the virus are in fact quite similar worldwide. The biggest impact of the epidemic, in
the bigger picture, is on worldwide trade. As countries all over the world are suffering from these virus
outbreaks, people worldwide are more ready to start to look at vaccinations even though their interests
might not be aligned.
Adjustments
No solution and strategy will decrease the risk of an outbreak to zero, and the virus will never truly
disappear. A good solution for the poultry industry should take into account animal welfare, food safety,
and economical considerations. Therefore, when thinking about our solution, we must take a
multistakeholder approach. Only when all these aspects are considered the relevant stakeholders are able
to actually implement the solution.
The stakeholder explicitly stated that vaccines themselves are not necessarily the problem from their
perspective, once a strategy for not disrupting trade will be found. However, there is still a big problem
with rapidly changing dominant serotypes, as well as discrimination between vaccinated and infected
animals and vaccine availability. The stakeholder states that we are doing good work filling the gap which
the industry now has: specifically preventing infection. There are two aspects of outbreak control: prevent
the virus from getting into farms, and prevent virus spread. Our solution could have an edge over
conventional vaccination in that sense.
The stakeholder also explicitly stated that it will be crucial for our solution to be effective alongside
already existing strategies. However, if we could further decrease disease spread together with
vaccination our strategy could really be something to research in the future. We certainly will keep this in
mind when thinking of future implementation and during our designing/engineering steps.
Our Next Steps
Think of how implementing our solution would work alongside already existing strategies.
Consider the multistakeholder approach described by the stakeholder in our solution.
Date of meeting: 27-09-2022
Description
We were interviewed by a someone from Nemo Kennislink who wanted to write an article/review on our
solution. This journalist also interviewed some experts in the field who could give their opinion on our
solution. One of these scientists approached us to discuss our approach further an in more detail. Since the
expertise of this scientist is very relevant to our project, we were very happy to have a conversation with
this stakeholder.
Contribution
We first discussed the scale of our research, and the specific experiments we are conducting to create a
proof-of-concept our solution. The stakeholder expressed that this strategy in theory looks promising, but
the real-world efficiency will greatly depend on whether our microbe is actually able to keep producing
nanobodies in the lungs for longer amounts of time, how long it takes for the nanobodies to diffuse over
time, and of course, how well they bind to the virus. In his experience, the most important factor usually
is the binding affinity of the nanobody towards different disease epitopes. After reviewing the article
where the nanobodies that we use are tested, the expert thought they had quite some potential because of
their ability to quite broadly neutralize different variants with good affinity. However, it will be crucial
that the nanobodies we produce with our bacteria also show these same results (or close to the literature
reported data).
Besides the specifics of our design, we talked a lot about how we could be able to continue our research,
even after the iGEM competition ends. Mainly on how we might be able to actually do in vivo testing in a
later stage of our research. Of course, only after we have a good proof-of-concept and genetically stable
GMO’s. We discussed the process on how to go about the designs and logistics of experiments we could
still carry out, and experiments which we could do in a further stage. Also, we discussed how we should
have a plan B. This is especially important if we would want to have our research funded later on in the
research and development stage. There is always a chance that our solution does not work, which means
that we would have to forfeit our plan. If we have a plan B, we could increase of receiving money to carry
out our research, as the risk of forcefully having to forfeit the project are smaller.
We discussed how we could also use nanobodies to directly vaccinate chickens by fusing together
nanobodies and hemagglutinin proteins. The nanobodies would in that case have to target MHC receptors
on dendritic cells, directly and specifically delivering hemagglutinin proteins to the cells that can invoke
an immune response. This has already been done in literature and shows very promesing results.
However, because of the nature of our solution, specifically not evoking a host immune response because
of trade regulations and the necessity to discriminate between vaccinated and infected meat, this
application is not relevant to us. It could still be a very good plan B however. In that case we could use
the same delivery platform (our Nanobuddies) to produce and deliver these nanobody/protein fusions to
immunize the chickens.
Adjustments
We will address the binding affinity of our L. reuteri (and/or E. coli) produced nanobodies by means of an
ELISA test against several Hemagglutinin variants. This to further provide a good proof-of concept.
We discussed safety to the environment, as well as safety for the chickens themselves. We have already
taken great steps to creating a solution which is safe to the chickens, by making use of a naturally
occurring lung microbe. To make our project more safe for the environment, we have created a two gate
kill switch which eases the problem of using GMO’s in the environment. The conversation with this
stakeholder again showed us the great (ethical) concerns when it comes to using GMO’s in the
environment. A good example which was brought up: ‘what if this system fails?’, ‘will the bacteria then
be allowed to spread freely?’. Because we make use of plasmids there is always a risk that these might
fail after multiple generations. Even though our kill-switch should be very effective according to the
literature on which we based its design, we again though on how we can further secure its stability within
our GMO. A stable genetic kill switch, of course means a stable, potentially highly effective
biocontainment strategy. The means by which we want to achieve a more safe and responsible solution to
the environment are by integrating our kill switch in the genome, rather than a plasmid. Since we are short
on time these considerations to the safety will likely only remain hypothetical however. Also, we briefly
discussed that in theory the GMO should get outcompeted by the wildtype bacteria after a certain time
period. The stakeholder expressidely stated that we should validate this theory by means of an
experiment, as this will both influence the efficiency of our solution, as well as greatly influencing the
safety of our solution to the environment.
Our Next Steps
Even though we are quite deep into the final moments of the competition, we will try to get into
contact with organizations such as Greenpeace to gain their perspective on our solution.
Provide proof for our theory that the GMO bacteria will naturally be outcompeted by the wild-
type bacteria (e.g. by means of studying growth curves).
October
Date of meeting: 01-10-2022
Description
Check out our survey and human practices page for a detailed explanation on the design, results, and
conclusions from the survey. Here, we will mainly discuss how the survey contributed to the final design
considerations of our solution, and briefly address the main findings.
The aim of our survey was to find out the following:
Public opinion towards avian influenza;
Public opinion towards vaccination in the animal industry;
Public opinion towards GMOs in the food/animal industry;
Public opinion towards our solution.
We hypothesized that the general public would not be too fond of our alternative protection strategy as it
involves GMOs in the food industry, something that often is not looked kindly on in the EU, as well as the
Netherlands. However, we also suspected that this might change for the better if the participant worries
more about avian influenza outbreaks, and would therefore consider our strategy earlier. We think that the
public opinion on vaccinations is tolerant, as we have just lived through a pandemic which taught almost
every inhabitant of the world about viral disease and vaccinations.
Contribution
First of all, we were surprised to see that the general concern about avian influenza is relatively low:
around 6 out of 10 on a 0 (not concerned) to 10 (very concerned) scale. This was also the case for
concerns about avian influenza outbreaks in the poultry industry specifically. In contrast to the interviews
we have conducted with stakeholders which were highly concerned, and considering the millions in
damages and millions of birds that die or need to be culled, we would have expected a higher number;
even though this number is still on the ‘concerned’side of the spectrum.
Our survey reveals that people in general are accepting of our specific solution, with only slightly higher
acceptance averages than conventional vaccination strategies. When combatting avian influenza
specifically, survey results showed that the participants were more accepting of disease combating
strategies, even if they would include novel strategies such as GMOs (although still slightly lower
compared to vaccination). Our survey did not reveal that people would prefer vaccination or a GMO
treatment when consuming animal products. Moreover, it does appear that people would prefer a GMO
strategy if it is cheaper and would provide for better protection against disease. We also did not identify
extra concerns about the presence of GMOs in meat deemed safe for consumption, when compared to
concerns about the presence of vaccines in meat deemed safe for consumption. Still, we were surprised to
see how high this concern was for both considering the fact that people are very willing to buy meat
coming from vaccinated/GMO treated animals; this was certainly not what we hypothesized beforehand.
Adjustments
Our survey revealed that our solution is not necessarily frowned upon by the participants. The biggest
consideration we will need to take into account is safety for humans and animals. Although our survey did
not necessarily reveal such conclusions, we realized that every question had these factors as an
assumption which might explain the positive responses. Therefore, we decided to add another dimension
to one main key-value which we have not addressed too much earlier on in our project: human-health.
Since our solution was focussed on preventing a next pandemic resulting from avian influenza, we
thought we had already addressed the human health aspect enough. As a result of our survey, we will also
take into consideration how the meat from animals treated with GMOs, might affect and influence human
health in a more literal sense: would nanobodies for example be able to make their way into the human
body?
Besides this, we identified that our respondents did not discriminate much when it comes to consuming
eggs or meat coming from GMO treated animals. As a result of previous interviews and discussions with
the team, we had anticipated that eggs would generally be preferred as we suspected that people might
feel more comfortable as there is another ‘imaginary layer’ between the animal and the animal product.
However, our results do not indicate such a discrimination, therefore we decided not to specifically focus
our solution on poultry that exclusively provides eggs.
Lastly, our results show us that it is important to people that our solution addressed a relevant and
important issue. When asked whether they would approve a GMO strategy to combat avian influenza,
participants in general answered higher approval scores than when asked the same question about disease
in general. This means that when implementing our solution, it is important to be clear that it is necessary
to address this epidemic, as that will most likely improve our initial approval ratings by the general
public.
Our Next Steps
Integrate another layer to our key value human health.
Rethink our plans to only implement our solution in egg-laying poultry, as we initially thought
that would be better perceived by the general public. This might be not that relevant when we
consult the results of our survey.
Establish in our implementation the relevance of our solution, with a particular focus on
combating an uncontrollable avian influenza epidemic.
References
[1] Ibañez, L. I., De Filette, M., Hultberg, A., Verrips, T., Temperton, N., Weiss, R. A., Vandevelde, W., Schepens, B., Vanlandschoot, P. & Saelens, X. (2011, 15 april). Nanobodies With In Vitro Neutralizing Activity Protect Mice Against H5N1 Influenza Virus Infection. The Journal of Infectious Diseases, 203(8), 1063–1072. https://doi.org/10.1093/infdis/jiq168
