Team:TUDelft/Human Practices/iHP
Integrated Human Practices
In the integrated Human Practices section, we elaborate on how interaction with stakeholders shaped our project. Make sure to also read how and why we interacted with stakeholders in Human Practices. Integrated Human Practices is displayed in two ways: in a timeline and in a story. The timeline tells you per stakeholder (or event) its main impact that led us to the creation of PHOCUS. The story ties all of our human practices efforts together by describing the stakeholders’ perceptions of the problem and the solution, ethical, legal, safety and security considerations and how we envision PHOCUS to be implemented in the real world (Figure 1).
Highlights
A summary of our main findings and how they influenced our project:
- We obtained a thorough understanding of the problem and desired solution for the desert locust crisis and the local context by talking to farmers living in Kenya and India, and locust control experts from national governments and the Food and Agriculture Organizations of the United Nations (FAO).
- We learned that current pesticides used against locusts are not safe for humans or other animals, or are not fast enough in killing locusts. Therefore, PHOCUS was designed to be both safe and fast. From thereon, PHOCUS was continuously tailored to the values and needs of its end-users, the FAO and national governments.
- Our first project idea did not involve killing desert locusts, but instead, was aimed at stopping the locusts from swarming. Multiple leading experts in the field of locust control told us that this may only make matters worse, and advised us not to use this strategy, and focus on killing locusts instead. Therefore, we dropped this idea and completely changed our project towards killing locusts.
- We investigated the public trust in top-down locust control and their perceptions towards the use of pesticides in Kenya and India through a questionnaire.
- We performed an extensive analysis of all safety and security considerations we could think of and discussed these with experts. These are described fully in Safe-By-Design.
- We gave a keynote lecture at a Locust Control Management Conference. Here, we promoted and discussed PHOCUS with leading locust control experts, NGOs, legislators and scientists. From this, we learned a.o. about the potential of locusts to provide nutrients to the people in food deprivation and decided to make PHOCUS safe for human consumption.
- We learned that, currently, locusts cannot be used as a source of food and feed because these could have an amount of toxic chemical pesticides. Therefore, by making PHOCUS safe for human consumption, the people affected by desert locust swarms could eat or sell these locusts to mitigate food deprivation or economic losses.
- Current locust control uses a standardized method of application. To successfully implement PHOCUS, it should adhere to these standards.
Our interactive timeline
Story
Perceptions of the problem
The first step in finding a solution for the locust crisis is to obtain a thorough understanding of the problem. By talking to multiple stakeholders we made sure we tackled all possible perceptions of the problem.
This year’s locust upsurge has heavily affected the country of Kenya. Therefore, we first spoke to Susan Nguku from the Kenyan Ministry of Agriculture. By interviewing her, we obtained knowledge about how locusts affected farmers and local communities in Kenya. Many people in Kenya had never seen locusts before in their lifetime. When these arrived, they were in shock. Susan Nguku told us stories of distressed farmers banging metal pots and practising black magic as desperate attempts to fend off the locusts. This merely scares the locusts away to the farmer’s neighbors, whose crops will then be destroyed, leaving the farmers scared and defenceless. Since farmers in rural Kenya are very difficult to reach out to, Susan Nguku helped us to overcome the distance and language barrier by asking Kenyan farmers to answer questions from our questionnaire. The people that responded to our questionnaire generally feel like they have no say in the fight against locusts, while they would like to have some.
We considered cultural and socio-demographic differences in the perception of the locust problem by also interviewing Shanu, who lives in the city of Jaipur, India and is an eyewitness of locust swarms. He told us that the people of Jaipur were scared of locust swarms since they have not seen it before. However, since people living in urban areas are not in agriculture, they are not directly affected. His relatives in Indian villages who make a livelihood from agriculture, on the other hand, have been severely impacted by loss of crops.
Next, we investigated the root cause of these crop losses and economic stability incurred by locust swarms with the aim of preventing us from curing a symptom instead of the real underlying cause. Ecologist and locust population dynamics expert Dr. Cyril Piou told us that locust swarms develop as a result of certain environmental conditions. In 2018, these conditions were a rare series of cyclones that hit the Arabian Peninsula. Unfortunately, it is impossible to tackle the real underlying cause of locust swarms as we are not able to control the weather. While the weather is influenced by climate change it remains uncertain if the recent rare weather event can be attributed to climate change. According to Dr. Piou and Em. prof. Arnold van Huis, climate change has nothing to do with it, while our contact persons at the FAO claim it does.
Perceptions of the solution
After considering the local context of the locust problem and its unsolvable root cause, the weather conditions, we investigated locust control operations and current methods that are used. From our conversation with Prof. Alexandre Latchininsky and Dr. Mamoon AlAlawi of the Food and Agriculture Organization (FAO) of the United Nations, we learned that all locust control is performed by highly trained professionals who either worked for a national government or the FAO itself. This is because the vast scale of locust upsurges and the costs of locust control makes it impossible for sole civilians to do it themselves. This was in sharp contrast with our impression of locust control, where farmers apply pesticides on their own lands. We acknowledged that it is not feasible nor suitable to have farmers apply pesticides on their own and therefore, decided that our solution must also be applied by the FAO and national governments.
Pesticides
Methods to prevent locust swarms that are used by the FAO and national governments include chemical- and biopesticides. These pesticides are sprayed on vegetation, which locusts will eat, or directly on flying locust swarms. According to Prof. Alexandre Latchininsky (FAO) chemical pesticides are toxic to other insects, livestock and humans and therefore, are not feasible from an ecological and human health standpoint on the long run. This was agreed upon by all stakeholders we interviewed regarding current methods. From Susan Nguku we heard that some people experienced itchiness after the chemicals were sprayed in an area in Kenya, but for unknown reasons no follow-up health monitoring took place.
Initially, part of our solution was to use dead locusts from the swarms as a source of food for the people who have lost their crops. In this respect, the toxicity and unspecificity of chemical pesticides is especially worrying because, according to former FAO locust expert Em. prof. Arnold van Huis, these locusts are very commonly used as food for people and their livestock but only if they are caught alive. However, Em. Prof. Van Huis told us that these locusts could have come into contact with a sub-lethal amount of chemical pesticide and hazardous traces could be present in their bodies. Therefore, we stopped our idea of using the dead locusts as food and paid extra attention to the safety of PHOCUS for human consumption.
An alternative to chemical pesticides is Metarhizium anisopliae, also known as Green Muscle. This entomopathogenic fungus is specific to locusts and safe for human consumption. However, according to the FAO and Dr. Cyril Piou, this biopesticide has not been used as much because it is too slow; it takes 1-2 weeks to kill locusts.
In conclusion, during interviews, the responsible authorities conveyed a dire need for a novel alternative that is safe for non-target species and fast-acting. Therefore, we decided to design PHOCUS in such a way it would fulfill these needs.
Operations
Locust control operations consist of monitoring the development of locust swarms and spraying the pesticides preventatively or as an emergency response. Dr. Cyril Piou and the FAO emphasized the importance of taking preventative measures as once locusts start swarming, they are very difficult to control. However, it is very difficult to prevent swarming as places where locust swarms develop are remote and unpopulated (FAO) and because coordination and collaboration of locust control between countries is difficult (Em. prof. Arnold van Huis). Political tensions and lack of funds makes it even more problematic (Em. prof. Arnold van Huis). Above all, when locusts have not attacked an area for a long time, people forget about the importance of preventative measures (Dr. Cyril Piou and Harold van der Valk). Hence, an emergency response is often required.
From Em. prof. Arnold van Huis we learned that, during an emergency response, the locust problem also becomes a political problem. If a country is "under attack" by locust swarms it is important for the national government to show they are doing all in their power to protect the livelihood of their citizens. This often leads to over- and ineffective- spraying of chemical pesticides. In addition, toxicologist Harold van der Valk mentioned that people spray anything that is laying around. This may even be pesticides that are not effective against locusts by people who are not trained to handle these chemicals.
Our initial project idea: degregarization
At the start of our project, our approach did not involve killing locusts, but was more animal-friendly. We wanted a more animal-friendly approach because the FAO and Dr. Cyril Piou emphasized that an alternative method to current pesticides must not eradicate locusts, seeing as these play a vital role in the ecosystem. Our initial idea was to let the infected gut bacteria produce an enzyme that degrades the locust pheromones guaiacol and phenol: β-decarboxylase. Because guaiacol and phenol contribute to the swarming behaviour of locusts, their degradation could prevent or hinder swarm formation. We came as far as having designed and constructed the corresponding BioBrick. However, both Em. prof. Arnold van Huis and Harold van der Valk warned us that this is not a suitable solution as we would actually make matters worse. They explained that the swarming behaviour of locusts is a strong advantage when one wants to control locusts. Swarms are highly concentrated, which makes spraying pesticides on individuals much easier. By preventing this gregarious behaviour we would dilute their concentration, but not their numbers. Thus, we would effectively create “super grasshoppers" spread out over a large area, which would need more demanding control. After these interviews, we decided to completely change our project and PHOCUS on killing locusts instead.
Ethical considerations
We learned from the FAO that those affected by locust swarms do not have a say in which pesticide is being used and that these people rely on the government and FAO to take action. This means that, if we want PHOCUS to be applied by the authorities, the general public would not be involved in the decision whether or not PHOCUS would be used on their land. However, we feel like this would take away some of their autonomy. Therefore, we wondered: Do these people trust these institutions in taking the appropriate measures? To answer this question, we reached out to Kenyan and Indian farmers. Results from the questionnaire to Kenyan farmers suggest that Kenyan farmers are distrusting of the FAO, if they say a new pesticide is safe while the opposite is true for national organizations. However, the small sample size prevents us from drawing conclusions. According to Shanu and the questionnaire of Indian farmers, the broader public of India does not question their government’s decision to use a particular pesticide and trust it when they say that it is safe. We acknowledge that locust control by civilians and famers is not feasible and PHOCUS should be applied by professionals. Nevertheless, according to responsible innovation principles, not only those who perform locust control operations should have a say in whether PHOCUS should be used, but also those affected by locusts swarms. This led us to define our Education and Public Engagement goal as follows: “Enabling the discussion, by education and inclusivity”. We, for instance, discussed synthetic biology, locusts and our project with Indian school kids through an interactive quiz. We also presented and discussed our project at the Virtual Practitioners Conference on Desert Locust Control Management 2020 with multiple professionals working at the frontline of locust control, both from science and civil society.
As PHOCUS uses state-of-the-art in SynBio, implementation could bear considerable consequences. Therefore, our project should be considered responsibly and ethically. To enable the discussion on the implementation, Frank van Steenbergen advised us to reach out potential proponents, such as NGOs advocating against genetically modified organisms, to understand their viewpoint, identify conflicts and come up with solutions for those. Sadly, these organisations did not answer our request. Dr. Zoë Robaey aided us in understanding the possible unique downsides of our product and emphasized how we need to set up mitigation plans for these downsides. This is a critical factor for obtaining acceptance of our technology. Therefore, we set up mitigation strategies that can be read in Safe-By-Design.
Susan Nguku also stressed that obtaining acceptance from the locals could be difficult as they have a strong association with viruses being harmful. Enabling the discussion on the potential benefits of viruses will be laborious but essential for eventual acceptance. Another unique downside was hypothesized during the meeting with Dr. Zoë Robaey: could we accidentally eradicate the locusts if PHOCUS works "too well"? With this question, we reached out to Dr. Cyril Piou, who reassured us that, without something comparable to a gene-drive, we would not be able to do this. While killing the gregarious locusts, solitary ones will always remain and consequently propagate their role in the ecosystem.
Safety and security considerations
After working on integrated pest management for the FAO for many years, Em. Prof. Arnold van Huis currently spends his time on promoting the use of insects as food and feed. This inspired us to use all the locusts we would kill using PHOCUS as a source of nutrition in the affected communities. Although locusts are already often used as food and feed in those communities, the FAO warns against eating locusts as these could be contaminated with chemical pesticides. However, Remco and Gerard Bod from Simonis B.V., a preselected pesticide supplier of the FAO, told us that people eat them anyway, simply because they have no other source of food. During our conversation, Em. prof. van Huis explained to us that locusts that are caught dead are generally not consumed, which made us discontinue our idea of recycling all the dead locusts as food. Nevertheless, worldwide implementation of PHOCUS could entail that living locust could be eaten safely, which would improve human and animal health throughout the affected areas.
As we find it important that our treated locusts could still be eaten by people and/or their livestock, we consulted Dr. Steven Hagens, who is an expert at using phages for food safety. He explained that all known lytic phages (in contrast to some lysogenic phages) are safe for consumption. As Dr. Zoë Robaey instructed us to look at novel side effects of our product, we analyzed the consequences of spreading the Cry7Ca1 toxin throughout the environment. Toxicologist Harold van der Valk saw no possible safety issues with using this toxin, as Bt toxins are already a regularly used form of insecticide in the environment. Nevertheless, adequate field tests have to be done to ensure this.
To consider the general safety of releasing an engineered phage in nature we contacted multiple experts on (engineered) phages. From these conversations, we learned that our approach was so novel that there was little to no comparable research on potential safety hazards. Therefore, we mapped all safety aspects of PHOCUS we could think of and researched these individually, including safety to humans, livestock, other insects, and the risk of PHOCUS mutating in any way, phage resistance by bacteria and pesticide resistance by locusts. A full list of our safety considerations during the development of PHOCUS can be found on our safety page. During a video call with Dr. Piers Millet from the iGEM Safety & Security Committee, we concluded that the best way to progress the safety of our project was to develop an approach of how we could experimentally test and validate whether PHOCUS would be safe to use in the real world. Our description of the tests that are required for gathering data needed to go from the lab to field trials can be found on our safety page..
Potential biosecurity issues were discussed with Dr. Zoë Robaey and Dr. Piers Millet. Dr. Zoë Robaey told us that technology can be misused, but there is a level of scientific literacy required to do this. Nevertheless, there is a need for safeguards that can only be designed once there is an understanding of how a product can be potentially misused. After this conversation, we had a structured discussion with the entire team on the potential misuse of PHOCUS using the guidelines of iGEMers Guide to the Future. According to Dr. Piers Millet, biosecurity issues arise from platform technologies, not from narrowly applied systems. This led us to the decision not to advocate our engineered phage as a platform technology, but instead, maintained the focus on its niche application: killing locusts.
Legal considerations
Looking into regulations and admission regulations, it became evident that there are only a few specific regulations for the use of engineered bacteriophages, although recently the field of medicine has shown great interest. This is a great example of the "pacing problem", where legislation and policy are lagging behind the frontier of innovation. This possibly creates both aggravating and hazardous situations. Dr. Cécile van der Vlugt, a senior risk assessor from the Dutch National Institute for Public Health and the Environment (RIVM), explained to us how this pacing problem stems from a lack of knowledge and data. But, with the right data, she told us that approval would be possible. Our lack of additional biocontainment strategy should thus not pose a problem, if we can prove our claim that it will not propagate in nature.
Implementation
When starting the project, we identified relevant stakeholders and their values and assessed to what extent these values were represented in our initial project idea. Our efforts towards a value-sensitive design of PHOCUS is described in more detail in Human Practices. From this analysis, we concluded that the value “accessibility” was not represented to a desired extent within the project design. According to Susan Nguku and Simonis BV, getting pesticides at the right place at the right time is difficult. Simonis BV told us that this is partly because of inert decision-making of the bureaucratic FAO. Logistic issues have become even more problematic due to hampered transport as a result of COVID-19. To solve this issue, Dr. Cyril Piou proposed the idea of decentralizing the production of PHOCUS. Dr. Zoë Robaey confirmed that decentralized production would aid in solving these logistic issues. However, she warned us that, if you do not train the people that will work for you, you are just using them for cheaper labor. Thus, if we want to produce PHOCUS locally, this has to be done in the right way so locals obtain knowledge, co-ownership and training. Dr. Piers Millet commented on our local production idea by saying that giving every community their own virus production facility raises safety, security, awareness and acceptance issues. Furthermore, Simonis BV told us that they do not recommend decentralized production, as this withholds us from assuring quality and safety of the product. If the quality of the produced pesticide is not of the desired level and something bad happens, the producer and supplier are held responsible. We concluded that, in this case, safety and quality are more important than accessibility and we dropped the idea of decentralized production. Simonis BV told us that they choose to produce their pesticide in the EU to ensure top quality. Therefore, the core values for our production facility are trust and reliability, through standardization and centralization.
Furthermore, we envisioned how PHOCUS would be used in the real world. Harold van der Valk told us that in locust control operations, small groups of people travel into the desert for multiple weeks to search for locusts. Although they have coolers, often these coolers reach temperatures of 20 degrees Celsius. Therefore, we concluded that for PHOCUS to be a suitable alternative, it would have to be sufficiently robust to survive in 20 degrees Celsius for at least three weeks. According to the FAO, an issue with biopesticides is that these are generally not able to withstand high temperatures and UV radiation. This problem can be solved by spraying pesticides on flying locust swarms directly. However, Em. prof. Arnold van Huis said that this would require planes to fly very near to a swarm, which can be very dangerous for the pilot. In addition, the bacteria we aim for are in the locust gut. Therefore, we concluded that for safety of the pilot and efficacy, PHOCUS has to be sprayed on vegetation instead of directly on locust swarms. According to the FAO and Simonis BV, micro-encapsulation methods exist to solve issues of instability in high temperatures and UV radiation. Micro-encapsulation methods have been used in combination with bacteriophages before. However, they also told us that fine tuning micro-encapsulation requires a lot of expertise and is beyond the scope of our project.
Our conversation with Simonis BV also shed light on the standards the FAO has for pesticide spraying on swarms. Pest controllers always spray 1 L of pre-mixed ultra-low volume pesticide per hectare of land. This way all the planes, nozzles and sorts can be used in the same way for all the different pesticides. It is important for PHOCUS to be also applicable complying to these standards as this would benefit the diffusion of PHOCUS as well as the ease of use and safety.
Finally, pesticide resistance can become a problem in the future if PHOCUS would be used extensively. Resistance may evolve because locusts obtain resistance against the toxins or because the bacteria obtain resistance against the phages. Toxin resistance was circumvented by incorporating two different methods of killing locusts (the Bt toxin Cry7Ca1 and RNA interference). Considering phage resistance, Dr. Steven Hagens told us that if we target at least two different receptors per bacterium, the chance of obtaining phage resistance is negligible. Thus, we decided that at least two phages need to be used per target bacterium.
Make sure to read why and how we interacted with stakeholders in Human Practices!
Dr. Franklin Luzia de Nobrega
Who?
Dr. Franklin Nobrega is a head of the Microbial Interactions Lab in the School of Biological Sciences at the University of Southampton.
Why?
The goal of this meeting was to discuss our idea of genetic engineering a bacteriophage to use it to turn the gut microbiome of locusts into a cell factory that produces toxins and kills the locust from inside out.
What?
- As proposed by Franklin, for the genetic engineering of our bacteriophages, we used the bacteriophage recombineering with electroporated DNA (BRED) protocol.
- The lytic property of the bacteriophages is part of our biosafety plan. When the bacteriophages kill all their hosts, they will die as well, because quorum sensing ensures that the amount of host bacteria always maintains a level that is sufficient for bacteriophage survival, we decided that it is safer to identify the genes responsible for quorum sensing in bacteriophages and remove them.
Susan Nguku
Who?
Susan Nguku is an Agriculture Extension Officer at the Ministry of Agriculture in Kenya.
Why?
To learn how the locust swarms affect the lives of people in Kenya and how the Kenyan government approaches this crisis.
What?
- Some people in Kenya have never seen locusts before. When these arrived, they were in shock. There was a nationwide warning by the FAO that locusts would come. However, in the local communities, there were rumors, but no real communication. Still, even if the message could reach these locals properly, they would not have the tools to do anything about the locusts.
- Locals revert to anything at hand to scare away the locusts from their land. Such as banging pieces of metal to black magic.
- Barriers for acceptance by locals include usage of a virus, as the term virus is associated with disease. Even though it might be beneficial that mindset is hard to get out of people's minds.
- Even with all the fears, some people would be interested if the pesticide is fully approved for safety by a governing body (such as ICIPE, KEBS and FAO) and if there is some lobbying for the product.
Shanu
Who?
Shanu is a resident of Jaipur, India where the locust swarms raided the city earlier this year.
Why?
To consider the cultural and socio-demographic differences in the perception of the locust problem by interviewing someone from another affected country.
What?
- Indian citizens trust the government when it comes to the safety of approved pesticides. Even more so if the government is responsible for the application.
- Since this was the first time the locusts had been to the area in decades, the city dwellers were only startled by the locusts but those who made a livelihood of agriculture were severely affected.
Omer Lavy
Who?
Omer Lavy is a PhD student at the Tel Aviv University of Israel at the Department of Zoology. His major research focus is on behavior and neural mechanisms of behavior in invertebrates, and specifically insect models.
Why?
Omer Lavy investigated the Effect of Density-Dependent Phase on the Locust Gut Bacterial Composition. In his publication, he characterized the microbiome of the desert locust.
What?
- As we could not confidently identify a certain bacterial species in the locust gut from literature, we decided to use the conclusion of Omer Lavy that the Enterobacter genus was widely present in most Locusts. Therefore, we decided to tackle the locust problem by creating a phage cocktail targeting the Enterobacter genus.
- In the dry lab, a key parameter is the amount of (target) bacteria that are present in the locust gut as this is directly correlated to the amount of toxin that can be produced. He, therefore, sent us some papers that quantified the number of bacteria in the locust gut, giving us a good estimate to use in our models.
Dr. Cyril Piou
Who?
Dr. Cyril Piou is an ecologist specialized in locust population dynamics, affiliated with CIRAD.
Why?
Experienced with locust control in Western Africa and has experience with the FAO.
What?
- Taking preventative measures against locust swarms is very important because these are hard to control once there is an outbreak. Since locusts tend to breed in remote locations, good forecasting methods have to be in place to understand where a locust can breed. Drones can be used to monitor and control the locust population in these remote areas.
- The relationship between media attention and locusts is cyclic. When there's a locust upsurge, the media pays attention to it. Media attention is important to obtain financial donations from other countries, such as France, to help set up locust control operations. The affected countries themselves are often not sufficiently responsive. The development of the logistics and practice of these operations continue for a few years until the locusts are gone and people don't find locusts a problem anymore. At that point, preventative measures are not being taken anymore and the upsurge returns.
- In the case of Green Muscle: Many people didn't know how to use it. Education of farmers needs to be encouraged so they understand how to use the biopesticide, the limited side effects, why it's much safer than chemical pesticides and why it's worthwhile waiting for 10 days for the locusts to die.
Em. Prof. Arnold van Huis
Who?
Emeritus Professor at Wageningen University. Currently spending all his time on the journal, Insects as food and feed. Worked on Integrated Pest Management projects from the FAO.
Why?
Worked on locust control in both the African region and Australia, including being the dutch representative at meetings concerning the desert locust.
What?
- People will only use locust as food if they are caught alive. The risk of potential health hazards is too big when they are plucked off the ground dead. Still there exists a health hazard if sub-lethal amounts of pesticide are sprayed on the locust, as these can be eaten during an upsurge. Making PHOCUS safe for human consumption would relieve this health hazard.
- Controlling locusts by returning them to their solitary state is infeasible as a control option, as now you have to deal with many individual and dispersed locusts.
- Usage of pesticides is a political problem as well. People want an answer to their problems that is quick. Efficiency and effectiveness of pesticide spraying in general is under some debate, as sometimes spraying does very little to a swarm, while changes in weather conditions often have huge impacts on swarms.
FAO CRC
Who?
Dr. Mamoon Alsarai AlAlawi, Executive Secretary for the FAO Commission for Controlling the desert locust in the Central Region (CRC). Cairo, Egypt.
Prof. Alexandre Latchininsky, Agriculture Officer at the Locusts and Transboundary Plant Pests Team (AGPMM) from the Plant Production and Protection Division (AGP)of the FAO. Rome, Italy.
Why?
FAO is the global authority responsible for locust control.
What?
- The FAO policy is that locust management should stay in the hands of educated professionals who are specifically trained to deal with locusts. Next to experts in locust control, the FAO works closely with governments. Governments lead the control of desert locusts because it is costly. Farmers cannot afford it. The Member States of the FAO trust that this organization chooses the right technology for control operations, pesticides and sprayers.
- We still rely on conventional pesticides and this is not feasible from an ecological and human health standpoint. There is a need to produce a pesticide that is fast, easy to use and environmentally safe. Metarhizium works but it is not the best solution. The main benefit is its specificity, but the drawback is that it takes 1-2 weeks to actually kill the locust.
- It is important to construct a biopesticide that is able to withstand high temperatures and UV radiation. Behavioral fever has been a problem when using biopesticides. When locusts are treated with the fungus Beauveria bassianaBeauveria bassiana, the locusts sense that they are infected. As a result, they position their body in a particular angle towards the sun, which increases the temperature and the UV radiation on the fungus. As a result, the efficacy was only 20-30% on the field, whereas it was high in the lab.
Harold van der Valk
Who?
Harold is an environmental toxicologist for over 20 years. Previously he worked with international organisations like the FAO, UNEP and WHO.
Why?
Early on, he worked with the FAO on pesticides and locust control. Currently he has his own consultancy bureau called Falconsult, once in a while he is still involved with locusts when it concerns health and environmental impact.
What?
- In an ideal situation locust control is done by small organisations. Small groups of people travel into the desert for multiple weeks to search for locusts, somewhat resembling guerrilla warfare. Although they have coolers, often these reach temperatures of 20 degrees celsius and we should keep this in mind during the design of our biopesticide.
- As locusts move a lot a specific area might be only sprayed once or twice. Leaving the effect of the chemical pesticide on the environment to a minimal level. This is an acceptable level concerning human health if this is done by professionals, who are properly trained and protected. If lay people use these techniques there are health concerns.
- There are concerns with the part of our project aiming to "de-gragarize" locusts. Harold is concerned we will effectively create "super grasshoppers". Solitary locusts are too spread out to target with pesticides, making control of them impossible. It is actually a virtue that locusts form dense swarms, as now there is concentrated target to spray. By degragerizing the locusts, the large number of individuals will spread out and reproduce, only spreading out the problem and possibly making it harder to control.
- There are many BT insecticides/pesticides being used around the world, producing these with PHOCUS should not pose a problem.
"the people tackling locusts need to be like locusts, who move quickly"
Prof. Jozef Vanden Broeck
Who?
Jozef Vanden Broeck, Professor in Animal Physiology and Neurobiology at KU Leuven, studies the physiological mechanisms implicated in the regulation of important post-embryonic developmental processes of insects, such as the phase polyphenism in locusts. In addition, he investigates the mechanisms of systemic RNA interference in insects to explore novel strategies for pest control.
Why?
We met with Jozef Vanden Broeck to obtain information about the gut physiology and microbiome of locust species. Furthermore, we were interested in insights we could provide in the use of toxins and RNA interference as pesticide. Finally, we reached out to him to discuss ethical considerations in using insects as a research object.
What?
- It is difficult to bring a toxin into the gut that would survive the gut conditions of the locust. Its enzymatic capacity is huge and the conditions during digestion are very harsh for having biological molecules to survive in.
- RNA interference works perfectly at very low concentrations in locusts if you can get it into the hemolymph. Locusts have evolved to have a very strong defense system against viruses because they are occurring in high density populations when they are swarming.
- According to Dr. Franklin Luzia de Nobrega (University of Southampton), bacteriophages survive in neutral conditions, somewhere between a pH of 6 and 8. According to Jozef Vanden Broeck, the pH of the locust gut is somewhere between 7 and 8, the gut is in terms of pH generally survivable for bacteriophages. This means that we do not have to take this into consideration anymore when deciding on the design requirements of PHOCUS.
- Real locusts are used in Jozef Vanden Broeck's lab, not cell lines. Regulations regarding conducting experiments using locusts are based on biosafety, not at the ethical level. Locusts are legally not considered in the bureaucracy like for mice. This stems from the fact that a lot of insect science is focused on killing them.
Frank van Steenbergen
Who?
Frank van Steenbergen is the owner of MetaMeta. MetaMeta is a value-driven company that is primarily trying to introduce solutions for water management. They try to do things that otherwise do not get done, i.e. challenges that are not addressed or overlooked also outside of water management.
Why?
Understanding the context of the affected areas from someone with local experience and contacts.
What?
- Frank feels very little is done regarding the locust upsurge, specifically the communication thereof. Media coverage is often superficial and not taking any initiative to find a solution.
- "The knowledge of the few can empower the many", Frank said to describe the current imbalance of knowledge between experts and the ones affected. He sees the lack of knowledge leads people to use ineffective measures.
- Considering implementation and opposition, Frank advised to get some of our potentially strongest opponents on our side as soon as possible. Integrating some of their needs and values into our project would see a lot of benefits regarding acceptance.
Dr. Zoë Robaey
Who?
Zoë Robaey is Assistant Professor in Ethics of Technology at the Philosophy Group of Wageningen University.
Why?
To discuss our project with her, so that we create our product with knowledge of our moral responsibility.
What?
- People work differently in different areas of the world - when a new technology is integrated in a different country it is important to think of the local context and not take how our countries are organised for granted.
- It is not enough to compare biocontrol to chemical pesticides in terms of problems that chemical pesticides bring. There is a need to study the new problem or solutions brought on by bio-control, for example, the effect on the ecosystem and other unintended side effects.
- A discussion with entomologists and/or ecologists would be interesting to know what ecological systems might be disrupted by these phages or killing large amounts of locusts. Therefore we discussed this with both Arnold and Cyril, the latter stressed to us that this would be impossible to eradicate locusts without the use of something resembling a genedrive. Arnold introduced us to a book describing the extinct North American locusts.
Dr. Piers Millet
Who?
Dr. Piers D. Millett is Vice President for Safety and Security at iGEM and co-chairs iGEM’s Safety Committee.
Why?
The aim of the meeting was to discuss all biosafety and biosecurity measures we took so far. Specifically, we wanted to validate whether we could say that our biopesticide is safe and does not raise any dual-use concerns.
What?
- The safety of our project relies on whether the Cry7Ca1 toxin is as specific as it is claimed to be. We know from two research papers that the Cry7Ca1 is specific to locusts, but how sure are we really? To ensure the specificity of the Cry7Ca1, it can be assessed by testing the selectivity and by performing field trials. Before we can do any field trial, we have to convince regulators to allow for these trials to happen on their ground. Therefore, first of all, we should consider which steps are between the lab and the real-world to find out which data we need to convince the regulator.
- We can back our argumentation that the genetically engineered phage does not require an additional containment mechanism by validating in the lab that, after some time, the genome sequence of the phage will return to wild type. This can be done by determining the genomic stability of our phage using serial passage experiments. It can also be used to assess whether the engineered system is sufficiently stable to be effective.
Prof. Jeffrey B. Jones
Who?
A distinguished professor of the University of Florida at the institute of food and agricultural sciences. Works primarily on strategies against bacteria that affect tomatoes, peppers and citrus fruits.
Why?
Acquired the first EPA registration for use of bacteriophages as biocontrol in agriculture.
What?
- That one of the primary concerns of biosafety concerning the use of wild-type bacteriophages in the environment is transduction.
- If a relatively large amount of toxin is required to kill the locusts, there might be a problem of selecting for locusts with a resistant gut microbiome. This is of little worry when little amounts of toxin are required.
- Bacteria become resistant to phages by a factor of one in a million or ten million. This is faster than resistance to antibiotics. To determine if this would pose a problem for our project, this question of resistance was modeled in the dry lab.
- If we can show that only the host of the phage is very limited, then the spread into the environment and the chance of propagating becomes smaller. This is also aided by the fact that bacteriophages outside of the locust are unstable. Therefore the targeted bacterium, or another in the host range of the bacteriophage, should not be present within the soil. Otherwise this would mean the bacteriophage is able to propagate, as within the soil the effect of heat and UV is reduced, increasing the stability of the phage while also enabling its propagation.
Dr. Steven Hagens
Who?
Chief Scientific Officer Food Security at Micreos, of which the mother company is PhageGuard (which uses bacteriophages for food safety).
Why?
He has been working with phages for 23 years, during his master, PhD and during employment at Micreos.
What?
- Bacteriophages do not make us sick or cause allergic reactions. It is true that bacteriophages shape the human microbiome. However, clinical trials showed that, even though participants obtained large doses of phages, it had no impact on their microbiota.
- Bacteriophages that integrate into the genome (lysogenic phages) are known to be able to make bacteria toxic and resistant. An example is the Shiga-toxin carrying phage. In contrast, non-integrating phages (lytic phages) have not been shown to be detrimental to humans in any way.
- The easiest way for a bacterium to obtain resistance is by modifying its receptor molecules. The best way around this is to use 2 to 3 different phages that recognise different receptors of the same bacterium. In case resistance becomes a problem this approach would be developed.
- The risk of bacteriophage legs mutating, whereafter another bacterium could be infected, is not very high, although it will not never happen. However, the bacteriophage legs have evolved for millions of years to be perfectly aligned to their hosts. Even if a bacteriophage manages to mutate and infect another bacterium, it will be competing with the natural phages.
Simonis BV
Who?
Remco and Gerard Bod are Export Manager and Director, respectively, of the Dutch agrochemical company Simonis B.V.
Why?
This company produces and exports insecticides and pesticides to many regions of the world, predominantly to the Middle East and Africa. It is a preselected pesticide supplier for, among others, locust control of the FAO for many years.
What?
- The FAO has standardised dilutions for all used pesticides, so that the same amount of volume is always sprayed on the same amount of land (1L per 1 hectare). In this way, the spraying practice and sprayer settings remain constant regardless of the active ingredient. Therefore it is important PHOCUS can fit into these standards.
- To be absolutely sure that the pesticide is of high quality, it is best to have it produced in Europe or the USA. Local production of pesticides in regions of Africa or Asia does not give sufficient assurance of the quality. The quality of the pesticide is not only important for its efficacy, but also to prevent any accidental side-effects. If something bad happens, the producer and supplier are responsible. Taking this into mind the idea of decentralized local production was dropped.
- The easiest way for a bacterium to obtain resistance is by modifying its receptor molecules. The best way around this is to use 2 to 3 different phages that recognise different receptors of the same bacterium. In case resistance becomes a problem this approach would be developed.
- The UN monitors the health of those involved in the application of pesticides and local community members. Although people are advised not to eat locusts because these may be treated with chemical pesticides, people will still do it because of a lack of other options to eat.
Dr. Cécile van der vlugt
Who?
Dr. Cécile van der Vlugt is a Senior Risk Assessor at the Dutch National Institute for Public Health and the Environment (RIVM). In that function, she works on risk assessment and risk management issues concerning genetically modified organisms. As an expert on new biotechnological developments, she participates in (intern)national projects on emerging risks and safety measures of these new developments.
Why?
To gain insight into the view of a risk assessor on our project and to understand how PHOCUS would fit into current legislation.
What?
- PHOCUS would need to comply with three different regulatory aspects; GMO regulations, insecticide regulations and food regulations. The latter surprisingly could pose one of the biggest hurdles, as there is the possibility of an association between the applied GM phages and plant cells, resulting in residual phages present in plants grown for food or feed. Although this is still uncertain and so are the risks, this should be further investigated.
- The need for an additional biocontainment strategy in the design, i.e. a kill-switch, would not be necessary as long as we have data to back up that our bacteriophage would not persist in absence of its target host. There will be more problems stemming from the general acceptance of GMO in or on food and feed. In Europe, only one GMO has been approved by risk assessment and not even policy-wise. This is less a problem in pharmaceuticals, like vaccines, so maybe the improved sustainability of PHOCUS could be the convincing factor for some.
- The use of a bacteriophage cocktail can lead to more regulatory complexity, as assessments and the likes have to be conducted for a biological entity (the phages) instead of a pharmaceutical (non-living). The lack of specificity from the cocktail can pose problems for risk assessment, but could also be used as a case study for these type of mechanisms for future applications.
- There is no gap in GMO legislation, rather a gap in data and knowledge. Cultivation of the phages could be difficult as this needs to be standardized, something that is currently very new and hardly done.