Team:Toulouse INSA-UPS/Implementation

Ghana Survey and ethical matrix for implementation on Earth

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Ethical Matrix on Earth

As a descriptive ethical matrix, its purpose is to highlight the elements that have influenced the decision-making process carried out by the team. Our aim is to build a project that respects and preserves the stakeholders. We also wanted a project that was as responsible and good for the world as possible. During the design of our system, three conditions were put forward based on the crossroads between stakeholders and values (well-being, autonomy, fairness):

• Material conditions: considering resources, to compensate for their unequal distribution

• Moral conditions: responsibility in design, responsibility of intermediary actors

• Cultural conditions: acceptability by users.

As a normative ethical matrix towards the team members, it is a way to constrain our social responsibility in this project.

Presentation of the actors involved in the development of our technologies for an application on Earth

Actors: any person or group of people who can decide about the right way to act and who can have an impact on that decision.

→ iGEM team

Users: people who use a technology and who may have certain wishes or requirements for the operation of a technology.

→ Population of vitamin A deficient countries, agro-industry and private companies

Regulators: organizations that formulate rules or regulations that technical products must comply with. These may include standards related to health and safety, but also guidelines related to competitor relations and fair trade.

→ Government of vitamin A-deficient countries and nutrition NGOs, Recycling and waste treatment actors

References:

[1] Van de Poel, I. and Royakkers L. (2011). Ethics, technology and engineering. An introduction.

Stakeholders	Autonomy	Well-being	Fairness
	The autonomy principle stipulates the need to respect the autonomy of an independent person who is considered to be free and capable	The principle of beneficence, which requires contributing to the well-being of others; Well-being is a state related to various factors considered separately or jointly: health, social or economic success, pleasure, self-actualization, harmony with oneself and with others.	The justice principle is concerned with the distribution of resources.
iGEM Team	Design transparency: the entire project, from the details of our design to the feedback on the experiments are detailed on our wiki, allowing any research team to take over our project or to use it as a basis to build their own project in complete autonomy.	Responsible design: our design respects the safety instructions for using and handling this system in the laboratory, by following the decree of July 16th 2007 (NOR: MTST0756429A, revised on February 16th 2018).	Fair access to information: all information about our project is freely available, accessible to all and well documented.
Agricultural industry and private companies
Actors in recycling and waste treatment	Right of review: these institutions have the right to supervise the design and implementation of the project.	Eco-responsible design: the design anticipates the end-of-life treatment stages of the device. Materials are studied to have a minimal impact after their end of life.	Sustainable production methods: we tried to design our system so that it is as resource-efficient and as easy to recycle as possible. In this way, we tried to create a more sustainable production method. However, some aspects still need improvement. Notably through the use of sensors in our device.
Vitamin A-deficient populations	Ease of use: Our device has been designed so that a non-specialized user, such as a resident of a country suffering from vitamin A deficiency, can use it without any outside help other than an instruction manual. This instruction manual could have been part of our future plans. We discussed this with the Ghanaian iGEM team with whom we collaborated during our interviews. Our device uses few resources and can be used for a long period of time. This means that, unlike a regular replenishment of chemically produced vitamins, our device allows people to produce vitamins by themselves:  this increases their autonomy. However, one of the scarce resources needed to use our device is electricity, and a survey we conducted among the Ghanaian population indicates that a portion of the population does not have access to electricity, which jeopardizes the use of our device in these areas. Additionally, the procedures for starting up and cleaning the reactor to avoid the release of GMOs into the environment require advanced equipment that is not readily available on site.	Deficiency complementing device: a future development of our project could be to verify and test the non-toxicity of the products and co-products generated throughout the production process. Tests to validate the safety of the device by operating in various and extreme conditions are also planned to prove the non-toxicity of our project regardless of the conditions in which it is used.	-
Government of vitamin A-deficient countries and nutrition NGOs	Right of review: these institutions have the right to supervise the design and implementation of the project.	Device solving a real problem, vitamin A deficiency in some countries: our process allows the production of vitamin A-enriched yeast using a minimum of resources and recycling them. Our process is therefore part of the actions carried out to limit vitamin A deficiencies.	Compliance with regulations: It is part of our plans, in the long term, to adapt our system to the regulations established in the various countries suffering from vitamin A deficiency. These regulations are numerous, they involve the configuration of the device itself or the health of the populations, with tests on the non-toxicity of the yeast.

Ghana survey on the acceptability and possible improvements of our system

We were thinking of implementing our vitamin production system on earth, in areas affected by vitamin A deficiency. Since Ghana is one of them, we needed Ghanaian population feedback in order to make our system evolve towards real life issues. To do so, we created a survey about the usefulness and acceptability of our system. In this survey, we asked for advice on how to improve the system according to the needs of the local population. Team AshesiGhana collected and analyzed the data of this survey.

This survey allowed us to highlight some limitations of our device for implementation on Earth. The fact that many people do not have electricity is a real difficulty knowing that our system uses electricity as a resource. Nevertheless, some solutions could be found to counteract this problem and the overall acceptability of our project for these populations is quite good. However, we were not able to survey a sufficient number of people for this survey to be representative.

A more detailed analysis of this survey can be found on the Partnership page.

Conclusion

The study of this matrix and this survey allowed us to understand that our device, entirely designed to be integrated in a spacecraft, is not suitable in its current form for an application on Earth for vitamin A deficient populations. The high price, the restrictive sterilization conditions are many points to address that make this application difficult. Moreover, one of our main resources is electricity, and as the survey carried out in collaboration with the iGEM Ashesi Ghana team has shown us, some populations do not yet have electricity.

Thus, our system will have to be rethought if we want to apply it on Earth. Many aspects of our project remain interesting, such as our nutritious yeast or the coculture system using minimal resources to produce compounds of interest.

Implementation

Overview

The implementation of our project consists of three modules. The first module is “Use of SynBio to solve an unmet need of astronauts”. We identified our potential customers and their unmet needs not yet covered by other existing solutions using a market study. The second module is “Implementation technical and sustainable constraints in our system”. We showed that our solution is feasible, scalable, and inventive through the specifications, the issue table, the 3D model and the life-cycle of our solution. The third module is “Supporting entrepreneurship”. This module is detailed on the Entrepreneurship page but we present here an overview and its main conclusions. We present logical product development plans with realistic milestones, timelines, resources, and risks. In short, we took an entrepreneurial approach to develop our project outside the framework of the iGEM competition. Throughout this process, an ethical approach is added around the implementation of our system for the space market. An opening will be made around the possibility of implementing our system in regions suffering from vitamin A deficiency. To see our ethical matrix for implementation on Earth, click here!

Our partners for the implementation of our project:

Le Catalyseur is a pre-incubator that encourages the culture of innovation, supports project leaders and enables the development of original activities in terms of employment, while maintaining high social added value. We followed their pre-incubation program which consists of three phases. We did 6 work sessions, one every month since May. They gave us tools such as market study to implement our idea in the real world and such as business models or risk analysis to have an entrepreneurial approach. The work that we have done with them is presented here. Now, we are at the end of the first phase and we have the opportunity to continue our project after the iGEM competition.

Toulouse White Biotechnology (TWB) is a catalyst weaving links between fundamental research and the industrial world. On the one hand, it offers collaborative public/private R&D projects and services. On the other hand, it supports start-ups in order to accelerate their start-up and growth. They support us financially and we had a meeting with them in order to be advised on the path to take. We also participated in the 2020 edition of TWB start-up day, which was on “Bioproduction for sustainability”. This is where we met Randy Rettberg.

Module 1 - Use of SynBio to solve an unmet need of astronauts

In this first part, we will see that our system has been built from the beginning to be implemented and produced, then we will study the market targeted by our device and we will establish the influence of our consultants in the positioning of our product.

First of all, it is important to note that we have started from a real society problem to establish our technology and not the opposite. This gives our device a real legitimacy and interest that makes its production realistic. We were able to establish this problem thanks to the expertise of Alain Maillet, a physiologist at CNES (French space agency), who helped us list the major issues to be resolved for long-duration space travel. That is how we discovered that vitamins degraded over time (to know more about it, see the description page of our wiki [here]). Doing so, we came to the problem of producing a nutritious yeast enriched with β-carotene to supplement the diet of astronauts during long-duration space missions.

Market Analysis

Segmentation

In order to define a marketing strategy and to know which market we want to target, we must first segment the market demand. We consider the market for products aimed at improving the health of astronauts for long-duration space missions. This is a niche market and as such we will have few segments.

Figure 1: Market segmentation

Targeting

The second step of the marketing strategy is to perform a targeting, i.e. to choose the segments on which we will focus. This targeting is done according to the size of the segments, their potential profitability or the ability of our company to meet the needs of these segments. Here, we will target space agencies with the potential to send manned flights into space and the SpaceX company.

• Estimation of segment size

• Correspondence between the need and the offer

Our product is a device for the production of β-carotene-enriched nutritious yeasts operating through coculture using minimal resources and aimed at supplementing the diet of astronauts for long-duration space travel.

Institutions aiming to send astronauts to space for long periods of time require fully recycled devices, using very few resources, to ensure the survival of astronauts.

Our product is a good match for the needs.

• Competitive analysis
• NASA Bionutrient Project: The BioNutrient experiment is part of NASA's SynBio project and will test a method of nutrient production in space that uses genetically modified baker's yeast to produce specific antioxidants, such as β-carotene. However, the system is not designed to be able to produce yeast indefinitely (unlike ours) but is in the form of sachets that can be stored like probiotics.
• VEG-01 NASA: This project aims to produce edible plants in microgravity.
• MELISSA ESA (European Space Agency) project: The MELISSA project consists in transforming a spacecraft into a closed ecosystem based on bacteria, algae, plants, chemical elements and natural processes.

Certain dimensions are more important depending on the project. Here, what matters most to buyers is that the product:

• Is sufficiently nutritious to supplement the deficiencies of astronauts
• Works in spite of the constraints of microgravity
• Is sustainable over time
• Consumes few resources
• Requires little maintenance and is autonomous
• Takes up little space
• Is not heavy
• Is sufficiently solid

We can then position the different competing authorities on the following diagram, according to the two most important values: the economy of resources and the nutritional quality of the product supplied. We chose these two characteristics because, out of all those mentioned above, they have the most decisive impact in terms of usefulness and difficulty of implementation.

Figure 2: Marketing positioning of our project

It can be observed that, with regard to the envisaged problematic, the iGEMINI project and the MELISSA project are those which best respond to the characteristics of interest on the market.
Indeed, in his interview, the former French astronaut Philippe Perrin highlighted the interest of our innovative project by declaring:

“What interests me in your approach is the nutritional capacity of your yeast.” , Philippe Perrin on the 1st of July 2020

If you want to learn more about this interview click [here]

Thus, to establish this market study we were able to rely, in addition to the literature, on the testimonies of two astronauts: Philippe Perrin and Jean-Jacques Favier as well as the testimony of Mr. Alain Maillet from CNES (French space agency) who helped us to identify the constraints that a device must respect to go into space. This allowed us to adapt our device to the constraints of the space environment and to those of long-duration space travel.
This market study gives us the direction to follow in our product development process to adapt the production methods, the business model and all the tools to the real market as much as possible. Indeed, in order to sell a product properly, it is necessary to know to whom to sell it and to know the characteristics of the market. This is all the more true when considering the space market, given the many constraints associated with it.

Module 2 - Implementing technical and sustainable constraints in our system

We demonstrated that our solution is feasible, scalable, and inventive through the specifications, the issue table, the 3D model and the life-cycle of our solution.

The system

Specifications

Space is a special environment with special constraints. We listed and answered the main constraints that our device has to meet in order to operate safely and meet our production targets. We have thought about many aspects, but sending a system into space for a long duration mission requires a lot of thinking on many subjects. We have thought about all aspects of our project, but there are still some things that need to be matured before we can launch our system into space (about the recycling of the medium, the degradation of urine for salts and urea, the system of light induction…).
All feedback from experts can be found on the IHP page [here].

Safety constraints

Constraint
The reactor must not contain a gaseous phase. This phase would not be at the top of the reactor as on Earth but all around the liquid phase. Evacuation of the gaseous phase is also complicated and it can be dangerous in addition to limit the gas transfer.

Our solution
Hollow fiber membranes can be used for the dissolution of gases. This method would allow us to regulate the dissolution of H₂, CO₂ and O₂ and avoid gaseous phase at the same time.

Experts who helped us in our choice
Arnaud Cockx, expert on process engineering, Head of the Transfer Interface Mixing team at TBI at TBI. Benjamin Erable, expert on biofilm engineering, member of BioSym department and biofilm engineering expert at the Chemical Engineering Research Center of Toulouse

Constraint
Local pressure variations should be minimized as it causes the vaporization of dissolved CO₂, H₂ and O₂ and the apparition of a gaseous phase.

Our solution
Variation of volumes and high flow can cause local pressure variations. Frontal filtration or piston pump should rather be used than peristaltic pump.

Expert who helped us in our choice
Christine Lafforgue, expert on biosystems and process engineering. She has already worked on a coculture system.

Yeast growth constraints

Constraint
Yeasts and bacteria must be separated, to not contaminate each other.

Our solution
We have thought of two reactors with medium flowing between the two through a filter.

Expert who helped us in our choice
Christine Lafforgue, expert on biosystems and process engineering. She has already worked on a coculture system.

Constraint
The medium must be homogeneous for a correct transfer of metabolites between the two microorganisms.

Our solution
There should be agitation in both reactors. The pumps need also to be adjusted for optimal medium recirculation.

Expert who helped us in our choice
Nathalie Leys, expert on biosystems and process engineering. She has already worked on a coculture system.

Constraint
The yeast must be harvested.

Our solution
We do not want the separation filtration system to be continuous. The yeast reactor should be purged every day with recirculation of the medium and the yeast should be collected on a washable filter and added to the astronauts meal.

Constraints for β-carotene production

Constraint
The system must produce 0.00279 mmol of β-carotene per day per astronaut.

Our solution
Thanks to the model, we know that this production can be achieved with the volumes of reactors below. The C. ljungdahlii reactor should have a size larger than 0.5 L to produce the desired amount of β-carotene in one day. Check our modeling page [here] to learn more about this.

Figure 3. Production time as a function of the reactor volume of Clostridium ljungdahlii and Saccharomyces cerevisiae

Induction of flavors constraints

Constraint
The cells should not receive outside light from because the flavor induction uses light.

Our solution
The reactor must be opaque. Stainless steel could be use.

Constraint
The yeast reactor must contain a blue and red light system that needs to be efficient enough to irradiate the entire reactor even with a high cell density.

Our solution
We did not have enough time to meet this constraint. Nevertheless, we know that the solutions put in place for photosynthetic microorganisms could be suitable for our problem.

3D model

Considering the specifications and the many feedbacks from experts, the 3D model of our coculture system was designed for space application. You can follow the path of our reasoning that resulted in the design of our 3D model [here]

Limits and possible improvements

After solving all the main technical issues, we used 3D modeling to get an overview of our system.

Obviously, this design can be improved and the system will meet new constraints when progressing in the Technology readiness level (TRL) steps [6]. You can learn more about TRL here. We already thought of an improved design with the help of Christine Lafforgue but we have to evaluate its advantages compared to the design we used for our 3D model.

Life-cycle and table of issues

Using the “Specifications” tool, we were able to list the technical constraints of our system and design an appropriate solution. We then planned the life-cycle of our system to have an overview of our product implementation in the real world. For each step of the life-cycle, we used the table of issues tool. This tool consists in listing the different problematics that could be raised from this step of the life-cycle (historical perspective, societal/environmental/economical issues, maintenance, safety). Thus, it is a useful way of thinking to build a sustainable, viable and responsible product.

Life-cycle

MANUFACTURING

Our product would be produced at a very limited amount, as we tackle a very specific and limited market (space projects). It would be only produced to respond to a specific need. We plan to outsource to an industrial company both the production of the reactor from the components listed (to a company expert in mechanical devices production, ideally oriented to space applications), and the production of our freeze-dried microorganisms to a company from the biotechnological industry. The choice of the materials used is critical as it has to resist space travel, and in particular the take-off step. We could collaborate with the CNES for this aspect. We plan to be involved in the assembling step of the reactor components produced, and to deliver our system as a kit containing both the reactor, the freeze-dried microorganisms and the culture medium.

TRANSPORT

Our system is transported directly to the spacecraft building place. During the transport phase, the freeze-dried strains must be carefully handled and keep them dried. At the end of the transport phase the product is integrated in the spacecraft.

USE

Our product will be used during space missions, in spacecrafts. The product is built to avoid heavy maintenance tasks for the astronauts using a monitoring system, and the astronauts will be trained before their mission to learn how to use the coculture system. We learned that from two astronauts interviews that you can find on our Integrated Human Practices page.

END OF LIFE

After a space mission, the GMO strain must be eliminated.

Table of issues

Materials and manufacturing

Historical perspective
Biotechnology related production (bioreactor, coculture, GMOs) is new in the space field and is at its very beginning. However, space exploration often accelerates the development of emerging technological fields. As biotechnology is considered a growing and innovative field of technology, it is expected that space exploration will take advantage of it.

Environmental issues
Our manufacturing process has an energetical cost, both for the production of the reactor and the microorganisms. However, the advantage of microorganisms, compared to a chemical process is that they could be grown at mild conditions (temperature/pressure) whereas a chemical process usually requires harsh conditions. Furthermore, the major resource to produce them are organic nitrogen/carbon sources and thus we do not rely on chemicals (e.g derived from petroleum). We can also note the fact that the production is not expected to be at a high scale as we are targeting a very limited and specific market.

Economical issues
Our system requires the use of hollow fiber membranes and several sensors for O₂ and CO₂ which are costly products. Research associated with the design of a biological reactor for space will require significant funding. However, spatial missions usually receive important endowment. We have to find financial support from private and public investors.

Maintenance of the system
We anticipate that several sensors must be taken, as they could stop functioning during a long space mission.
The materials that make up the flexible parts of the system, such as the hoses, must be thoughtful to have a lifespan longer than that of the space mission.

Safety
The absence of any manufacturing defect is particularly important for space projects. The technical maintenance in spacecraft is limited to the materials and the skills on board. A small technical problem on the device could therefore have heavy consequences on the spatial mission. Thus, our manufacturing process requires the highest level of quality check. Also, specific safety rules regarding GMOs production must be integrated.

Use

Historical perspective
Nutrition has always been a concern for a successful exploration, more specifically since the period of great discoveries during the 15th century. The first vitamin isolated and used for nutrition was produced at the beginning of the 19th century.

Societal issues Providing a solution for vitamin deficiencies in space while it is still a big problem in some parts of the world could be criticized. That is why we considered adapting our technology on Earth, as developed at the end of this page. The use and ingestion of GMOs is also still debated. One solution could be to neutralize the yeast before ingestion.

Environmental issues
Using microorganisms in space could potentially contaminate new planets and compromise the search of extraterrestrial life. Clear instruction of the use and disposal of the coculture system should be given to astronauts during training.

Maintenance of the system
The astronauts should be trained to autonomously handle all the possible issues they may encounter while using our system. The necessary maintenance needs to be as little as possible. The system and filtration is designed to delay clogging of the filter. However, it would still be necessary to provide for regular purges to change the filters. This raises the question of sterility.

Safety The main risk of use is contamination of the reactor that could affect the health of astronauts.

End of life

Historical perspective
In recent years, there has been more awareness of the issues related to the “end of life” of technologies (societal, environmental, economical and even spiritual). Materialism and the short life of many technologies are now heavily criticized. However, this issue has poorly been integrated in space projects since the beginning of the spatial conquest and there must be an improvement in the field. As a new innovative project led by young biological engineers, IGEMINI must open the path towards a more responsible space exploration.
Another solution could be to eject the reactor into space before returning to Earth. This solution is questionable in relation to the fate of the waste in orbit around the Earth, of the waste burning in the atmosphere and of the microorganisms in space.

Economical issues
Providing a solution for vitamin deficiencies in space while it is still a big problem in some parts of the world could be criticized. That is why we considered adapting our technology on Earth (cf Little Cosmos on the bottom corner). The use and ingestion of GMOs is also still debated. One solution could be to neutralize the yeast before ingestion.

Environmental issues
It is necessary to assess the additional costs that the recycling of faulty parts will generate.The release of the reactor into space would not cost anything.

Safety
GMOs produced for our technology must be eliminated at the end of the space mission, using appropriate sterilization solutions.The release of the reactor into space does not pose a safety issue.

Using life-cycle analysis and tables of issues, we raised 3 key points that are critical for the technical development of our IGEMINI project.

1. GMOs: concerns about GMOs were noted at each step. GMOs must be on one side demystified for citizens, and on the other side carefully handled (containment and elimination).
2. Training and maintenance: we noted that using an innovative technology in space is challenging as the astronauts cannot rely on technical support. Thus, a deep quality check is essential before using our product, and the maintenance must be reduced as much as possible. Furthermore, training must be given to the astronauts on several parts of the system: avoiding risks, handling the maintenance tasks and the use of GMOs.
3. Sustainability: even though we do not plan to have huge production rates (our product will be only used by astronauts), we realized while using these tables of issues that we had some responsibilities. Thus, we decided to consider finding a second life for our reactors after a space mission, and to think about how we could use and adapt our system for vitamin deficiencies on Earth in developing countries.

The iGEM’s aim is for student to get scientific knowledge and lab experience. These are not the aims of the development of a start-up. We realized that there are many concerns to handle because of the table of issues. We did not handle them for the iGEM competition because we have preferred to manipulate and reflect on biological problems. The start-up iGEMINI will need to address all the issues raised in this table.

Ethics, a key point in the implementation of our system

Throughout our implementation process, we considered the issues that our project could raise, in terms of economic, societal and environmental aspects. This approach is supported by our ethical matrix for space, which allowed us to highlight the impact of our project on its stakeholders regarding the values of autonomy, well-being and justice. The work around the Sustainable Development Goals has also allowed us to show that our project is part of a more sustainable development. Our project is innovative, responsible in its production and consumption methods, contributes to the well-being of the astonauts and involves many partnerships. We have thus proved the feasibility of our project from an ethical point of view in space applications. This is unfortunately not the case for the application we wanted to make on Earth (cf. Little Astronaut on the right side). Thus, our device will have to be rethought if we want to apply it on Earth. Many aspects of our project remain interesting, such as our nutritive yeast or the coculture system using minimal resources to produce compounds of interest.

Whether in the treatment of plant effluents or in the complementing of deficiencies, our project remains a project for the future, both in space and on Earth, and it is up to us or our iGEMeur successors to make it a reality.

Module 3 - Supporting entrepreneurship

In this part, we present product development plans with realistic milestones, timelines, resources, and risks. In short, we took an entrepreneurial approach to develop our project outside the framework of the iGEM competition.
We collaborated with a business incubator named “Le Catalyseur” which helped us in this process through workshops throughout the project.

Here are the steps that we have done for an entrepreneurship approach:

• Objectives and missions of the company
• Strengths, weaknesses, opportynities and theats
• Risk analysis
• Business model
• Business plan
• Milestones for space application

Go on the entrepreneurship page to follow our approach in more detail [here]

After these analyses, we know that our technology is valuable. We have several opportunities to develop our technology for space application or for other applications.

Our project got the attention of the Vice President R&D Operations and Innovation from Pierre Fabre. He advised us to continue to develop our technology as a start-up and he thinks that we could be hosted by the Fondation Pierre Fabre. We still have to define the mutual benefits of such a collaboration. The production of vitamins or other nutritive agents could be of interest for them.

Toulouse White Biotechnology is a cluster whose members share common social-economic goals in relation to industrial biotechnology. TWB also hosts and supports starts-up. As our partner, they are an opportunity for technological transfer to other applications.

The CNES (French National Centre of Space Studies) is with the Catalyseur, our main partner. The CNES launched the project Space Ship France to find solutions to space constraints for interplanetary ships. With the Spaceship project the CNES proposes to build a place of innovation, open, unique and immersive, facilitating collaborative and interdisciplinary work, learning and experimentation to better federate, stimulate, innovate and promote French excellence in the fields of Exploration and Human Spaceflight. Its objective is to facilitate the exchange of information between experts in the field and projects under development in order to boost the projects and allow them to mature faster.

We explained the aim of the Catalyseur in the beginning of this page. This is a free public help for students for the development of innovative projects and starts-up. We followed their pre-incubation program which is available for three years. We finished the first year and the first step, the state of play. We can continue this program to learn how to make this iGEMINI project a company.

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Interview with Brigitte Godard, ESA doctor, you can read the full interview here

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