Team:TUDelft/Entrepreneurship/Product

PHOCUS

Product

After obtaining a thorough understanding of the people, the local context, and the customer needs, a Product Description was derived. We investigated the risks involved with its development and introduction in a Risk-Benefit Analysis.

Product Description

As previously described in Customer Needs, the FAO and national governments are in urgent need of an alternative to current chemical and biopesticides that is fast and safe. We offer these customers PHOCUS: a biopesticide that is specifically aimed at desert locusts to reduce the threat these insects pose to food security and economic stability. The active ingredient of the biopesticide is engineered bacteriophages. Adjustments of the phage enable the production of compounds that are specifically harmful for locusts.

An overview of how PHOCUS works is provided in Figure 1. The biopesticide can be sprayed on vegetation to prevent locust swarm development or as an emergency response against ravaging locust swarms. The biopesticide ends up in the gut of the desert locust through eating the vegetation. Here, the biopesticide transforms the gut bacteria of the desert locust into little factories. These factories produce compounds that are specifically toxic to the locust, not to other animals or humans. These compounds have two lethal functions:

  • It punctures the gut wall
  • It stops essential functions in the desert locust
Figure 1. Overview of functionality of PHOCUS.

The active ingredient, the bacteriophage, is useful as a delivery system because:

  • Bacteriophages can only infect bacteria, which benefits the safety of PHOCUS.
  • Only a small dose of bacteriophages is required to induce a large effect. This is because a bacteriophage is self-replicating inside the locust gut.
  • It can easily be taken up by the desert locust through ingestion. This way, PHOCUS gets to the location in the locust body where it is most effective: the gut.
  • Genetic engineering strategies were applied to induce maximum toxin production by the factory bacteria.

The use of two different toxins has two main advantages:

  • The toxin that pokes holes in the gut enforces the functionality of the toxin that halts essential functions in the desert locust.
  • The dual mode-of-action prevents the locust from becoming resistant against this pesticide. Pesticide resistance is a common problem in the field of crop protection.

Two main measures through which the safety of PHOCUS is ensured are:

  • The use of bacteriophages. These can only infect bacteria, not humans, animals or plants. Also, the type of bacteriophages (called lytic phages) do not induce an allergic reaction nor a toxic effect in humans.
  • The toxins that are produced inside the locust can only specifically harm locusts.

For a full description of all safety measures, please see Safety.


Customer Value

The market for the biopesticide PHOCUS contains two customer groups: the Food and Agriculture Organization (FAO) and national governments of areas that are affected by desert locust swarms. The FAO is responsible for locust control operations and aids national governments in their efforts in reducing the swarms. It provides national governments with a list of recommended and permitted pesticides and funding to purchase these.

PHOCUS creates value for these customers through its:


  • Superior functionality

The FAO and national governments of the affected areas are in urgent need for a locust control method that is safe and specific (to prevent harm to the environment and the health of humans and animals), and that is fast-acting (to reduce food insecurity and economic loss) exists. Current pesticides are chemical pesticides that are fast but unsafe, and biopesticides are safe but slow (for a full description: see "Current Methods on the Market"). Therefore, regulators and locust control operators in invasion areas are forced to choose between imposing the well-being of their people by exposure to chemical pesticides or by food deprivation. The biopesticide PHOCUS represents a unique combination of safety and speed that would relieve the customers from this dilemma.


  • High quality

The FAO, which has a strong voice in which pesticides will be used by them and national governments, highly values the quality of the pesticides. Therefore, in the manufacturing of PHOCUS, emphasis is placed on quality assurance. In practice, this means that the manufacturing of PHOCUS will be done in Europe because the European Union has high quality standards for manufacturing that is internationally recognized, and that the company will be involved in every step of the process to perform quality checks.


  • Compatibility

Current pesticides are being applied through spraying in ultra-low volumes in a highly standardized manner. For every pesticide, 1L has to be applied to 1 hectare of land. To ensure compatibility with current equipment, the biopesticide PHOCUS will also be sprayed in ultra-low volume in which 1L treats 1 hectare.


  • Radical innovativeness

Beside a unique functionality, PHOCUS represents a radical innovation in the field of crop protection. The concept of bacteriophage engineering is emerging and a bacteriophage-based pesticide is extremely novel and innovative: it has never been done before.


Risk-Benefit Analysis

The risk-benefit analysis is split up into four sections: risk analysis, a mitigation plan, a sensitivity analysis and a risk-benefit balance. In the risk analysis, potential bad effects concerning the introduction of PHOCUS were identified and ranked based on their probability and impact using a risk matrix. A mitigation plan for these risks was developed. The outcomes of these risks were predicted in a sensitivity analysis. Finally, these risks were weighted against the benefits of introducing PHOCUS in the final risk-benefit balance.


Risk Analysis

The main risks concerning the introduction of PHOCUS that were identified include:

  • Inefficacy of PHOCUS in killing locusts
  • Unintended effects of PHOCUS
  • Misuse of PHOCUS
  • Legislative opposition
  • Barriers to technology diffusion
  • Lack of interest from industry


These risks were ranked based on their probability and their impact from a scale of 1 to 5, which is displayed in Figure 2. The higher the risk scores in these two respects, the more priority it should be given in constructing a risk mitigation plan.


Figure 2. Risk matrix of main risks concerning the introduction of PHOCUS, positioned based on their impact and probability of occurrence.

The explanation why a risk has been positioned in a particular way on the risk matrix is described below.

More information on risk matrix

Inefficacy of PHOCUS in killing locusts

Different reasons for inefficiency or inefficacy were identified. For instance, an ineffective supply of bacteriophages to the locust, e.g. too little phages get into the gut, or the damage of the phage by UV radiation. Also, there could be a slow spread of the bacteriophage inside the locust gut as a result of harsh conditions or impaired replicative ability. Furthermore, the bacteriophage host bacterium might also be absent in the locust gut. Other reasons could be ineffectiveness of the produced toxin, too little production or undesired aggregation. Additionally, the shRNA may not be able to reach its target by being unable to cross the gut epithelium. Finally, an impaired release of the phage from its encapsulation could also pose problems concerning the inefficacy.

Probability: 2
Although many efforts have been made to develop PHOCUS and trials should have been completed before implementation, still, we cannot rule out that something will not work as intended. This is an intrinsic quality of dealing with the complexity of nature.

Impact: 5
If there is a dependence on PHOCUS to work, there could be an extremely devastating impact. For instance, this could lead to no other pesticides being on site or even produced at all. If PHOCUS fails to be effective, there would be no safety net, resulting in locust swarms that will devastate crops.


Unintended effects of PHOCUS

Releasing an engineered bacteriophage into the environment comes with concerns. Currently, no known methods exist to retrieve such an engineered entity. Different ways to prevent unintended spreading should be incorporated in the design thereof. But this can not fully rule out that it could have some effect on the environment that was not intended, which is inherent to the complexity of nature. Some questions arise to better understand the problem. How long will it be present in the environment? Will it be able to propagate or will it infect anything else?

Probability: 2
Although expert opinions have displayed the instability of phages within the intended environment, it will not remain a total negligible probability.

Impact: 3
Take note that this is the current outlook on what natural occurring systems it will disrupt. This is very difficult to assess because of the inherent complexity of nature. It could be the case that unanticipated systems will react or react differently than we had anticipated.


Misuse

Three factors make this technology vulnerable to intended misuse: the possibility of employing this technology for other purposes, the upsurge of biohacking [1], and the lack of legislation on bacteriophages and the engineering thereof [2]. This technology can also be misused without maleficent intentions due to a lack of understanding of the technology, its risks and implications. Another identified issue revolves around the fact that access to the biopesticide could be used as a political bargaining tool, i.e. only granting access if certain other requirements are met.

Probability: 4
Because of the many different ways in which the technology can be misused, as identified above, there possibility that any of these events will occur is quite high, but single events are already a lot lower.

Impact: 2
Many of the events described will not have a direct detrimental impact on society, although they may contribute to overarching more impactful movements.


Legislative opposition

Due to the novelty of this technology, legislation is not well-established [2]. Although the use of bacteriophages as an antimicrobial tool is common within Kenyan poultry farms [3], regulations on the modification of bacteriophages are unclear. Possible legislation imposed by governmental bodies may limit the development and implementation of the technology. Opponents of making genetic modifications, like GreenPeace, will also lobby for legislative opposition. Making it very likely that opposition will be met.

Probability: 5
People are precautious of the unknown, especially in the european union where this is core to their legislation. Meaning there will be a lot of opposition as much as unknown. Also taking into account this concerns a genetically modified entity, even more opposition will take hold. Coming together this comes to the highest probability.

Impact: 5
If parts of or the whole project become illegal, PHOCUS would not be able to be used.


Barriers to technology diffusion

End-users may be discouraged from using this technology due to a lack of knowledge and awareness of the technology or its application, the lack of complementary goods (for example, compatible spray- ing systems or storage units), insufficient financial resources, socio-cultural aspects, language barriers or unfavourable institutional aspects. Even though most compounds and the application thereof is primarily led by the FAO, national governments still dictate what happens in their own countries while the FAO advises.
These choices are primarily made in the Agriculture ministries, as they are often responsible for locust control. These problems are not only relevant on an individual level but also on a national level.

Probability: 2
If PHOCUS works as it has been designed, having an advantage over other alternatives, the chances of this are small. Still something, like the use of genetic modifications, might result in objection from local communities. Their chance is there but it is slim, people are desperate for a solution to the locust problem.

Impact: 3
Impact mostly revolves around those involved with PHOCUS, for others life will go on as it was. Still being affected by the Locust swarms though.


Lack of interest from industry

For large-scale production of the biopesticide, collaboration and cooperation with the industry is favourable. Companies possess financial resources, manufacturing facilities, control distribution channels and have expert knowledge. The risk exists that the firm may not be interested in this biopesticide because production might be expensive. While the product should be cheap as many of the affected countries are poor. This might leave a small or even negative profit margin.

Probability: 3
Without a clear view of how big profits are going to be made, companies are often careful with their investments. This would be seen as a risky investment and currently the resources are absent to convince them otherwise, so this is quite probable to occur.

Impact: 2
The decentralized production would still be able to take place even without industry interest, but it should be noted that this will make balancing the financials more difficult.


Mitigation Plan

Inefficacy of PHOCUS in killing locusts

To mitigate this risk, robustness and reliability of the technology must be high. To achieve this, genetic stability, by identifying the best location for the genetic insert, and encapsulation could be used, therefore improving environmental safety.


Unintended effects of PHOCUS

As posed in the description, retrieving the bacteriophage after release will be impossible. Different measures to reduce its chances of unintended propagation should be incorporated into the design. The first important factor is fitness, often the naturally occurring bacteriophages are robust and optimized for surviving and propagating itself, "the survival of the fittest". In our project genetic information is altered and introduced in the phage, reducing its fitness (Franklin, interview). This will leave them at a disadvantage, as the introduced elements will not form any competitive advantage over wild type phages. In theory the phage will be outcompeted and " go extinct".

Bacteriophages also require hosts to propagate, if these are not found in a relatively quick timescale they will also "die". Phages are relatively unstable in the phyllosphere [4].

Bacteriophages are extremely specific to their target bacterium, making it improbable that they will infect unintended targets but never impossible. Another aspect of this issue is that the targeted bacterium might also be present within the gut of other local animals. This would open up the opportunity that other animals can be "infected", but because the toxin is so specific it should have little effect on them [5]. This should also apply for humans, although a toxicity report would be required before implementation to ensure this.


Misuse of PHOCUS

To make it harder to repurpose our solution with malevolence it should have a high level of specificity towards locusts. Even though it might be modular in the sense that different bacteria can be targeted, the produced compounds within infected cells should be specific. In that case, changes to the system would have a marginal impact and repurposing the technology will take a lot of effort and expertise.

Collaborate with policy makers on how to deal with this risk. Advise responsible parties such as national governments to impose regulations and sanctions when said regulations are breached. Furthermore, educate end-users about the implications and direct uses of the technology to prevent unintentional misuse.


Legislative opposition

Adopt lobbying strategies to influence decision-making as often opposition arises from a lack of knowledge. If people understand, they can rationalize what they see as acceptable or usefull, while the unknown often scares people away. It is also of great interest to reach out to said opponents to acknowledge their opinions and adjust our project accordingly, if possible.


Barriers to technology diffusion

To increase knowledge and awareness of this biopesticide, details on the phage construction, production and delivery should be transparent and accessible through practices of open science and by having a clear website. Moreover, education of the public on the benefits and risks of our bio/pesticide compared to the current pesticides and bio/pesticides is of great importance.


Lack of interest from industry

As the possibility exists that our biopesticide will be disrupting in relation to other pesticides in the industry, other companies would be inclined to invest in our technology and safeguard their market share. It should be noted that it is geared at the current use of preventive biopesticides for locust control. Dr. Cyril Piou pointed out to us that making loads of pesticides for swarms once every so many years is of no economic interest to industry as this is not a profitable business plan in the long run.


Sensitivity Analysis

In this sensitivity analysis, we predicted which consequences might result from the occurrence of risks previously mentioned in the risk analysis. This tells you something about which factors may influence the success of the implementation. This is of great importance in making the assessment whether our product is likely to produce value for the local companies.


Inefficacy of PHOCUS in killing locusts

  • If control efforts rely on PHOCUS and it does not kill the locusts, they are free to do and go as they please. In this case they will swarm and spread. Ravaging crop and pasture all around.
  • The locust could develop a form of resistance to the produced toxin, rendering PHOCUS ineffective. Not many specific toxins are known so replacing it with another is something difficult. But our complementary approach of combining RNAi with the Cry7Ca1 toxin should render resistance development almost impossible.

Unintended effects of PHOCUS

  • Permanently propagating GMO in the environment which we are unable to remove.
  • Target bacterium of the bacteriophage might be under bigger selective pressure and might be washed out of the environment of the phage.
  • The gene encoding the toxin might end up in different places, it is known that phages play a role in horizontal gene transfer [6]. These bacteria might now be able to produce this toxin.

Misuse of PHOCUS

  • As safety measures are put into place to prevent unintended spread of the bacteriophage into the environment. Misuse could lead to unintended proliferation of PHOCUS in the environment, because these measures might not be effective. This could also be the case in areas where PHOCUS was not initially intended to be used and the interactions with the environment are unknown, resulting in unintended impacts on the environment.
  • Misuse could also be based on malicious intent. In this case PHOCUS could potentially be used to infect bacteria in the gut of humans and produce a protein toxic to humans. Although this would require laborious efforts and knowledge, PHOCUS could still be seen as a spark lighting the fire.

Legislative opposition

  • If legislation prohibits the use of PHOCUS the current state of affairs would remain.
  • If the procedure for legalising the use of PHOCUS is lengthly, it could happen that another locust upsurge takes place in the meantime.
  • If PHOCUS gets prohibited because something lacks, i.e. safe production procedures, it could prohibit similar products from entering markets as the general outline of the idea might become against the law. Which means this type of solution might not be able to solve other problems.

Barriers to technology diffusion

If the technology does not get picked up by the end-user the current state of locust control will remain, with all of its shortcomings. The pitfall is that in those cases the technology has been developed, and that PHOCUS contains sunken costs.


Lack of interest from industry

This could lead to cases where PHOCUS cannot be produced as there are too little funds to start production. In addition, PHOCUS would then not be developed after the iGEM period.


Risk-benefit balance

Benefits of applying PHOCUS compared to contemporary methods.

  • Faster than current biopesticide

A big drawback of the currently used biopesticide is the time scale in which it kills the locust. This is between 1 and 2 weeks. This is especially problematic when trying to control swarms. As in this timeframe they are able to ravage enormous plots of lands, while also still being able to lay eggs. Our solution is aimed to have a shorter time of action, making it a more effective tool for swarm control.


  • No effect on human health, when using a lytic phage

Our project aims at using lytic phages, which do not possess a threat to human health as explained by Steven. This is in contrast with chemical pesticides, which do affect human health like Fenitrothion [7]. Which results in a requirement of protective equipment and health monitoring of those handling the pesticides.


  • Toxin is specific to locusts

The toxin is specific to locusts. This toxin is then non-toxic to other animals or insects, so even when either their gut bacteria are infected or consume a killed locust it does not pose a threat. On top of that the toxin is a protein. Meaning it is susceptible to being degraded and becoming unstable, preventing accumulation in nature.


Conclusion

The benefits of the application of PHOCUS are assumed to be 100% true for the sake of comparing the risks to the benefits. Currently, the responsible locust authorities have displayed a serious need for a novel pesticide against desert locusts (FAO, interview; Cyril Piou, interview), which is safe and fast. Something that PHOCUS brings to the table. There are some risks that could cause great harm, i.e. propagation in nature or malicious misuse, that would not be seen as acceptable when weighing the risks and the benefits. Therefore, these risks should be further assessed to give more detail on their severity, which helps risk managers to determine their acceptability. The other risks, although potentially detrimental to further use of PHOCUS, would be worth taking when compared to the benefits for human and environmental health.