Working in a new dimension in synthetic biology, we also believed it was necessary to questionate ourselves about the philosophical perspective of synthetic microbial communication.
As a descriptive ethical matrix, its purpose is to highlight the decision process done by the team at every step of the project in the respect and integrity of stakeholders. During the conception of our device, three conditions are emphasized from the crossings between stakeholders and values (well-being, autonomy, fairness):
material conditions: taking into account the resources, to compensate their unequal repartition)
moral conditions: responsibility in design, responsibility of the intermediate actors )
cultural conditions: acceptability by users.
As a normative ethical matrix towards the members of the team, it is a way to compel our social responsibility in this project.
Presentation of the stakeholders in the development of our technology
Actors: any person or group of people who can make a decision concerning the right way to act and who can have an impact on that decision. As actors, we also wanted to introduce non-humans, in our case the environment.
→ iGEM Team, environment, non stakeholders, future generation
Users: people who use a technology and who may formulate certain wishes or requirements for the functioning of a technology.
→ Users local, NGO
Regulators: organizations who formulate rules or regulations that engineered products have to meet. It can be norms concerning health and safety, but also guidelines linked to relations between competitors and fair trade.
→ NGO, government, WHO
Ref.: Van de Poel, I. and Royakkers L. (2011). Ethics, Technology and Engineering. An introduction.
Remains to be examined
Currently being examined
Already examined or inherently true
No control possible for now
Click on the numbers to see how our different matrices evolved with the life cycle of our device (see their descriptions below)
1
2
3
4
5
6
7
8
1- Brainstorming
2- Production
3- Sales
4- Education
5- Distribution
6- Containment
7- Utilization
8- Waste treatment
Well-being
Autonomy
Fairness
iGEM team
Technological and innovative challenge; responsible design
Design choices; no ideological/financial constraints
Respond to a worldwide problem
NGO
Users
Inclusion in the conception process
Transparency of design
Improving everyday life
Environment
Early risk management
Respect of biodiversity
Eco-friendly by design
Government
Safe in accordance with public policies
Transparency of design
Social inclusion
WHO
Non stakeholders
No long term negative impact
Transparency of design
No long term inequalities
Future generations
No long term negative impact
Discussion about synthetic biology
No long term inequalities
Well-being
Autonomy
Fairness
iGEM team
Quality control; respect of the specifications
Choice of technical production solutions
Respect of environment and labour in the production process
NGO
Respect of their quality and traceability norms
Collaboration; possibility of inspection
Users
Quality and performance controls
Fair production price
Environment
Eco-friendly process; use a cluster structure to limit the transport between the factories (ie device production, Membrane production)
Durable production process; value protection of ecosystems as much as workers
Government
Employement of workers in the production process
Right of review
Respect of the state legislation in terms work condition and GMO
WHO
Right of review
Non stakeholders
No impact on their quality of life
Future generations
Constant optimization of the production process
Review the production process when new technologies
Well-being
Autonomy
Fairness
iGEM team
Acceptance of product by stakeholders
No commercial constraints; freedom to offer product
Fair evaluation of product by stakeholders
NGO
Proven added value compared to existing solutions
No lobbying; freedom of choice
Fair price; homogeneity in price around the world
Users
Product integrity at delivery
Discussion about the product, inclusion in the NGO's choice
Distribution to different NGOs in the world
Environment
Easily transportable; reduction of transport pollution
No disturbance of natural ecosystems
No unnecessary transport
Government
Right of review
Financial help of governments towards the best existing solution
WHO
Proven added value compared to existing solutions
Right of review
Distribution to different NGOs in the world
Non stakeholders
Future generations
Evaluation of long term business plan, viability of the project
Freedom to change for another solution
Regular evaluation of selling price
Well-being
Autonomy
Fairness
iGEM team
Responsiblity of explaination
Elaboration of educational supports
Right to expose the considered use of the device
NGO
Able to transmit general messages along with the speciifc explanations of the device
Participation in the elaboration of educational support; veto power about the message transmitted
Even distribution of educational support
Users
Useful key elements about cholera and use of the device: education done in accordance with cultural approches
Education availbale at all time
Equal access to education; comprehensible by everyone
Environment
Education done about the respect of environment
No contradictions with the preservation of the environment
Government
Education integrated with the one already in place
Veto power about the message transmitted
No competition with education done by the government
WHO
Supports also explain basic hygiene rules
Inclusion in the elaboration of the user manual, to work together towards a better health
Information about the existing structures to help the population
Non stakeholders
Trustworthiness in the information given
No right to deliver an information as general truth without debate
Future generations
Long lasting information passed generation after generation
Upgrading of educational supports
No detrimental health information in the long run
Well-being
Autonomy
Fairness
iGEM team
Ensure the disponibility of product; alternative distribution pattern
Right of review
Fair distribution according to the initial project
NGO
Product integrity at delivery; no disturbance of routine activities
Possibility of re-discussion of the collaboration; decision of distribution
Responsability in the even distribution of the product
Users
Product integrity at delivery; accessibility of product
Discussion about the product; inclusion in the establishment of the distribution process
Fair distribution; not discriminatory
Environment
Easily transportable; reduction of transport pollution
No disturbance of natural ecosystems
No unnecessary transport
Government
Right of review
No countries excluded by the distribution
WHO
Distribution to populations in need
Right of review
Fair distribution; not discriminatory
Water treatment firm
No disturbance of daily life
Non stakeholders
Distribution does not create long term inequalities between populations affected
Autonomous gestion of the distribution without the help of NGOs
Constant evalutation of the distribution pattern
Well-being
Autonomy
Fairness
iGEM team
Scientific responsibility; global managing of containment respect
Decision in the scientific solution kept to achieve containment
Homogeneity of containment in production through time
NGO
Care responsibility; evaluation of containment
Right to review their collaboration if containment not satisfying
Homogeneity of containment during storage and distribution
Users
No danger in drinking the water
No intervention needed; emergency infromation accessible
Homogeneity of containment in the different distribution patterns
Environment
No spoilage of surrounding water
No disturbance of natural ecosystems
No leak of containment, no destruction of natural organisms
Government
No degradation of valuable natural ressources; no negative impact on population health
Right of review
Fair access to containment verifications information
WHO
Respect of health recommendations in place
Right of review
Right of review
Non stakeholders
No degradation of environment
Right of review
No negative impacts
Future generations
No long lasting negative impacts
Re-assessment of containment scientific solution
No degradation of valuable natural ressources
Well-being
Autonomy
Fairness
iGEM team
Efficiency, trustworthiness
Right of review
Use understandable by everyone
NGO
Easy to set up; trustworthiness
No need of external help to use it
Care for everyone
Users
Efficiency; trustworhtiness; acceptability = no change in customs (water taste, color....
Easy-to-use, no disturbance of professional activity
Understandable by everyone
Environment
Maintaining biodiversity; maintain heterogeneity of ecosystems
Respect of natural ecosystems
Non-human life valued as much as human lifes
Government
No impact on the economical activities of its population; positive impact on the health of the population
Right of review
Adapted to the country at stake in terms of language, culture, traditions
WHO
Impact on the worldwide cholera epidemics
Right of review
Right of review
Non stakeholders
No interference with their quality of life
No interference with their quality of life
No interference with their quality of life
Future generations
Adapt the utilization to new scientific datas discovered
Proceed to vigilance over time of possible secondary effects of the use of the device
Well-being
Autonomy
Fairness
iGEM team
Care responsibility
Right of review
Waste treatment solution examined by authorities and NGOs
NGO
Simple waste treatment procedure, integrable in existing procedures
Not too much constraints, liberty to end the collaboration at anytime
Not to be responsible for all the costs
Users
Protection from risks
No need of external help to discard used device; inclusion in the setting up of the procedure
Equal access to this waste treatment; spatial and social repartition of the waste treatment
Environment
Maintaining biodiversity; reducing transport of waste
No disturbance of natural ecosystems
Non-human life valued as much as human lifes; help with the treatment of other wastes
Government
Integration in actual procedures
Right of review
No additional costs for treating the wastes
WHO
Respecting health recomendations concerning GMO waste
Right of review
Right of review
Non stakeholders
Not affecting the integrity of natural ressources in the process
No interference with environment
Solution for other wastes as well
Future generations
Not affecting the integrity of the ressources in the process
Autonomous and long term waste treatment solution
Solution available if needed
1 - Brainstorming
This ethical matrix is the one describing the important points taken in consideration to choose the subject. As an iGEM team we wanted a technological and innovative challenge, to surpass ourselves. We also wanted to help resolve a concrete worldwide problem, having a positive impact on social and environmental aspects.
2- Production
The key element emerging from this ethical matrix is that our production process needs to be as eco-friendly as possible. Thus applying all the classical rules of production as follows: use a cluster structure to reduce the transport, re-use facilities, treat production waste properly, choose the most optimized solution for each step of the process, from plant heating to final packaging. As the technologies are evolving rapidly, is seems then obvious that all the decisions taken need to be re-assessed regularly to ensure that the processes in place are the more efficient ones in terms of resource comsumption and secondary products production. Another important point is the respect of the workers, we need to make sure that the health standards for working in the factories will be respected at all times during the production.
3- Sales
The total freedom of choice and transparency of our selling process is intrisinc to our apporach and poses no problem. For the future, we have to work on communication around our product to be sure that the stakeholders know what is it and can take an enlightened decision. These stakeholders will then also be included in the setting of commercial deals. Then to respond to most of the challenges of this matrix, a business plan and a SWOT analysis were carried. Currently, an evaluation of the final price of the product is carried to ensure its long term economic viability in comparison to existing systems offered to NGOs. Furthermore tests are done to ensure the added value of our product. A problem that still exists for this step is that for now we have only contacted NGO doing actions in the African continent and thus thought our device to fit in their organisations. To further develop our product we need to ensure that we are not creating inequalities in the world, and making our device accessible et each place where cholera is a problem. For that we need to widen our contacts and think of adapting the selling process to different NGOs.
4 - Education
Here "education" describes the fact that with the distribution of a new product and the autonomous use of it, there is a minimum required education to do. Explaining to the local population the right way to use it is part of the responsibility of developping a new product. To be able to reach the attention of the population, this education needs to be done in accordance with cultural approaches and will introduce environment preservation. For example currently, theatrical representations are done to easily explain hygiene gestures to the african population. This education needs to be understandable by everyone, whatever their country, education or language... it has to be universal. On the other hand, the NGO participating in the device distribution need to be included in the elaboration of the educational supports since they will be in the foreground of this education. As a result of this matrix, it was decided to create a first draft of a user manual. It will evolve with the advices of NGOs and the consultation of local population to best fit their needs. To ensure an easy and fair access to this education support, it is necessary to always combine it if the device and make it available anywhere it is used.
5- Distribution
The distribution process was thought as a step carried by NGOs as they are the closest intervenant to the populations at risk and over the years they have established a trusting relationship with them. A key feature of this step is that these NGOs must have the freedom to re-evaluate the distribution pattern, as it must not disturb their routine activities and as they are in the best position to know the real needs of populations. On the other hand it is essential to ensure that the distribution process is helping populations in need, fairly and in a non-discriminatory way. For that the multiple stakeholders need to have of review on the distribution process and in that case NGO can no longer carry the distribution by themselves, other instances need to be able to replace them. This distribution can be re-evaluated regularly by demand of any of the stakeholders. For the future the ideal situation would be for the populations to self-manage the repartition of the product. In the meantime we have to make sure that the distribution pattern does not create long term inequalities between populations.
6 - Containment
Containment is a key element of our project and more generally in a lot of synthetic biology project. As these projects rely on modifying microorganisms, they also have to ensure the safety of the environment by not introducing non natural organisms in the wild. For that purpose, containment needs to be a priority at each step of the life cycle of the product. It is thus a responsiblity for many stakeholders in the process. Leakage of the container would result in spoilage of the surrounding area, possibly leading to an imbalance in its microbiota. A leakage in the drinking water would also lead to the contamination of the drinker, situation for which significative researches need to be carried. In more concrete terms, each stakeholder needs to verify the containment of the product. the iGEM team during production and sale, the NGOs during storage and distribution and local supervisors need to be appointed to manage storage and use. This containment constraint also needs to be managed during explainations of the system to the populations, they have to understand the importance of it. Therefore in the instruction manual considered, information concerning the verification of containment and emergency behavior to adopt in case of problem need to be clearly stated. The majority of the challenges concerning containment are still being adressed because it is not acceptable to do a rush job on them. Moreover, since the global system was not obtain in the laboratory in the time of the project, final tests were not carried and are still to be done.
7 - Utilization
As already stated in the education ethical matrix, in order for everyone to ba able to know the right way to use the device, a user manual needs to be done. The main challenges of utilization are technical and social but both are mutually dependent. To have a system user-friendly, trustworthy and acceptable depends on the technical choices made. The first key element that needs to be emphasized is the user-friendliness, without this users will not use it and will not even want to try it. Acceptability comes after, if the population tries it, the first uses will determine if it will be accepted or not. Acceptability relies on not changing the habits of the users, meaning that the utilization should not interfere neither with any kind of professional activity, nor with their daily life routines. The taste, color and odor of water needs to remain unchanged. Trustworthiness is dependent on the quality of our system and its result in its real-life utilization, if the populations knows how to use it and accept it, they will judge its trustworthiness on the long run. If our system gives them good results and no one falls sick using it, its trustworthiness will gradually increase. We can also try improve this trustworthiness feeling by showing NGOs and local people of influence the scientific results to convince them of the efficiency of our device. They will then induce trust in the users. Furthermore for now we do not have the benefice of hindsight and thus we need to conduct regular assesment of the social and technical balance between benefices and risks of the use of our device. This continual evaluation needs to be done with the help of the NGOs and the population, reporting to us problems encountered. The kind of surveillance will allow us to coninually adapt the use of the device and correct potential flaws.
User manual disclaimer:
The notice was made to provide a pragmatic support to the use of the Bluepuri bag project.
The notice was established with hypothetical data. The duration of the water treatment was estimated thanks to modeling. We are aware that complementary results are needed to validate the main features of our system. We set up the response time at 1 hour according modeling work.
In the context of our social justice engagement, we also produced a more symbolical user manual which aims at being understood beyond the language barrier and to guarantee a fair understandability and a good utilization of our device. This user note is supposed to be distributed to NGOs and any potential user. Making two different user manuals was needed to adapt to different users. They also ensure traceability in our process.
8 - Waste treatment
We firstly imagined a simple waste treatment solution comprised of a container where people can throw the used device, the NGOs were imagined as the responsibles for bringing and collecting these containers. Considering the contamination risk this solution needs to be more elaborated and people need to know the right gestures to treat their waste, education needs to be done and as for the utilization, a user manual needs to be done. Furthermore this treatment solution can not only rely on the NGOs, the population should be able to operate the waste treatment itself. The goal for the future being that the population can treat the waste completely autonomously. Speaking of waste treatment for our system we realised that we should also think about the treatment of other waste in these countries. It led us to the idea that if we want to have a great impact we have to take into account the current situation of waste treatment in the country targeted. For instance in Africa in spite of the efforts engaged in waste treatment there are still problems of transportation, storage and treatment1. We should thus think of a way to adress this situation at the same time as we imagine the waste treatment system for our device. This also applies if we want a fair access to the treatment solution, inequalities in waste treatment already exist and need to be adressed first if we want a fair access to the waste treatment of our device.
Croc’n Cholera makes it possible to think of a new ethical dimension about genetic information and communication in synthetic biology. The discovery of quorum sensing gives rise to a lot of questions but we emphasize a philosophical perspective, because communication is fundamental in the world.
The discovery of quorum sensing
For many years synthetic biology has been interested in engineering only one organism. The task assigned to this organism can be quite diverse: protein production, biosensor… The recent work on quorum sensing (QS) communication systems can be qualified as a small revolution in synbio because it allowed the identification many mechanisms responsible of a lot of bacterial behaviors: biofilm formation and toxin production for example.
Homo sapiens tried to study QS system and to turn it to his advantage: e.g. quorum amplification encourages the biofilm formation and is particularly studied in the mining industry to improve the copper chelation power of bacteria2 . Quorum quenching is used in order to inhibit the biofilm formation3 or the expression of virulence genes. Medical applications are numerous as well as industrial projects e.g. to reduce membrane biofouling4
With a few molecular backbones, AHL and CAI-1 for instance, a lot of combination of communication molecules were produced and were viewed as as many language elements to interact with bacteria and modify their comportment.
But can we truly speak about language? What defines the language, can it come down to a panel of molecules?
The strength of quorum sensing of communication
In nature, at the wild type state, we can wonder about the status of a bacterial biofilm. Several works show that in this structure each bacterium has a specific role and acquires it by communication with its pairs and by sensing the environment5
. Can we compare these well-organized bacterial structures to other remarkable structures in wild life? Like anthill, termite mound, beehive or the human political society (only if we are thinking with the analogy between biology and collectivity in the thoughts of G. W. F. Hegel (1817), Claude Bernard (1865, 1867) and Georges Canguilhem (1968)) which are characterized by the construction of a superior edifice and by task repartition. The main difference is that in a biofilm the bacteria form the structure and there is no matter moving. But we can all remember the impressive picture of “ant rafts” during hurricanes in south United States. In this case the individuals directly take part in the formation of a structure.
Nevertheless, the formation of such structures all have one thing in common: communication. But with bacteria, due to the opposition between pluricellular and unicellular organisms, it is more impressive: can a biofilm be considered as a punctual pluricellular organisms like fungi or fruiting bodies?
Those examples demonstrate the power of communication and language in wildlife: from a single isolated organism an incredibly powerful structure can be developed. At the end, all the cells together are superior to the single sum of each cell by giving better survival chances6 .
We can also think of another mechanism: symbiosis and more specifically commensalism. Communication is tight with common metabolism cascades and contact interactions. The symbiote quantity is well balanced to ensure the equilibrium of both organisms.
Synthetic communications and their potential downward slides
Considering the strength of an interacting system, we can clearly ask ourselves the question the outreach of a synthetic communication between two organisms that do not normally communicate together.
To our minds, no works report any issues about synthetic communication. Nevertheless, we carried a reflection about our synthetic consortium and the potential downward slides of communication.
According to the literature the word consortium refers to cooperation between minimum two stakeholders in order to realize a task. In our case, the players are a cholera mimicking E. coli, V. harveyi the sensor, and Pichia pastoris the effector and the protein producer. The final task being to produce antimicrobial peptides (AMPs) in order to eliminate V. harveyi and E. coli upon the detection of QS molecules. So the consortium task is to inhibit 2 of its own members. At the end, we intend the eradication of both E. coli and V. harveyi. To effect this task communication is presupposed. In our consortium, communication molecules play a double role: sense the target and transmit the information. Special attention must be given to the choice of the signaling molecules. Leaks and their consequences have to be considered and investigated.
The communication molecules are specific (CAI-1) or are not usually meant to communication (diacetyl) so the leaks are relatively limited. We can assess that no communication molecules will be interfering with external organisms.
Consortia for microbial treatment do not show controversial consequences mainly because the population of microorganisms is heavily regulated by the selection pressure induced by AMPs. There is still the resistance phenomena that can advantage a microbe, which will became problematic. But this feature is common to every antimicrobial substance.
Considering these facts, it is hard to establish a pertinent snapshot or catastrophe scenario for our consortium.
The problem can mainly be due to the engineering of our effector, the main features of bacterial life and natural selection are competition with other organisms to access the resources and adapt to the environment. By engineering Pichia pastoris to produce antimicrobial peptides killing bacteria (and more specifically Vibrio sp.) the risk is to destabilize the natural equilibrium. That is why we need to ensure that this effect is contained in the device and does not affect natural water sources.
Perspectives of synthetic consortia
With the mastering of communication systems, the range of possibilities for microbial consortia seems to be very large. The use of microbial consortia aims to simplify the genetic engineering work to build a complex synthetic system. “Natural microbial consortia hold many appealing properties in one bioprocess, such as stability, functional robustness, and the ability to perform complex tasks”7
However the applications have to be studied with an introspective and prospective approach.
The misuse of this technology can be real: e.g. design of a BSL-3 organism consortia with synchronized virulence genes.
Philosophical opening
Finally, can communication redefine the definition of an organism or the barrier between the self and nonself?
On the field of hubris, the human can affirm itself a geological force capable to pressure evolution by engineering. By using genetic engineering the organisms can be specialized (or differentiated) and their proportion can be regulated by synthetic circuits (see Imperial College 2016). Those mechanisms can imitate (more or less) true biological developmental phenomena (like apoptosis and cell differentiation).
Can this artificial design meet the complexity of life and still unknown mechanism?
Even if synthetic biology gives us plenty of tools, we are far to create the first pluricellular organism based on the association of different microorganism species.
In this perspective, the Integrated Human Practices and Public Engagement aspects will need to be at the center of our reflection8 .
We may have discovered what a gear is, let’s now build a watch!
References
Lin, L. and Meighen, E. (2017). BACTERIAL BIOLUMINESCENCE. [online] Photobiology.info. Available at: http://photobiology.info/Lin.html [Accessed 1 Nov. 2017].
http://photobiology.info/Lin.html
Weiland-Bräuer, N., Kisch, M., Pinnow, N., Liese, A. and Schmitz, R. (2016). Highly Effective Inhibition of Biofilm Formation by the First Metagenome-Derived AI-2 Quenching Enzyme. Frontiers in Microbiology, 7.
https://www.ncbi.nlm.nih.gov/pubmed/27468282
Weerasekara, N., Choo, K. and Lee, C. (2016). Biofouling control: Bacterial quorum quenching versus chlorination in membrane bioreactors. Water Research, 103, pp.293-301.
http://pubs.acs.org/doi/abs/10.1021/es303995s
Jiang, L., Zhou, J., Quan, C. and Xiu, Z. (2017). Advances in industrial microbiome based on microbial consortium for biorefinery. Bioresources and Bioprocessing, 4(1)
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5306255/
Evolving information in living systems, a pathway for the understanding of cooperation and other major transitions, Livio Riboli Sasco, 2010
https://www.theses.fr/155130935
